HSPA12A Attenuates Lipopolysaccharide-Induced Liver Injury Through Inhibiting Caspase-11-Mediated Hepatocyte Pyroptosis via PGC-1α-Dependent Acyloxyacyl Hydrolase Expression

Liver dysfunction is strongly associated with poor survival of sepsis patients. Cytosolic lipopolysaccharide (LPS) sensing by Caspase-4/5/11 for pyroptosis activation is a major driver of the development of sepsis. Studies in macrophages and endothelial cells have demonstrated that LPS is inactivated by acyloxyacyl hydrolase (AOAH) and leading to desensitizing Caspase-4/5/11 to LPS. However, little is known about the cytosolic LPS-induced pyroptosis in hepatocytes during sepsis. Heat shock protein 12A (HSPA12A) is a novel member of the HSP70 family. Here, we report that LPS increased HSPA12A nuclear translocation in hepatocytes, while knockout of HSPA12A (Hspa12a−/−) in mice promoted LPS-induced acute liver injury. We also noticed that the LPS-induced Caspase-11 activation and its cleavage of gasdermin D (GSDMD) to produce the membrane pore-forming GSDMDNterm (markers of pyroptosis) were greater in livers of Hspa12a−/− mice compared with its wild type controls. Loss- and gain-of-function studies showed that HSPA12A deficiency promoted, whereas HSPA12A overexpression inhibited, cytosolic LPS accumulation, Caspase-11 activation and GSDMDNterm generation in primary hepatocytes following LPS incubation. Notably, LPS-induced AOAH expression was suppressed by HSPA12A deficiency, whereas AOAH overexpression reversed the HSPA12A deficiency-induced promotion of LPS-evoked and Caspase-11-mediated pyroptosis of hepatocytes. In-depth molecular analysis showed that HSPA12A interacted directly with peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) and increased its nuclear translocation, thereby inducing AOAH expression for cytosolic LPS inactivation, which ultimately leading to inhibition of the Caspase-11 mediated pyroptosis of hepatocytes. Taken together, these findings revealed HSPA12A as a novel player against LPS-induced liver injury by inhibiting cytosolic LPS-induced hepatocyte pyroptosis via PGC-1α-mediated AOAH expression. Therefore, targeting hepatocyte HSPA12A represents a viable strategy for the management of liver injury in sepsis patients

Acetylation of Myocardin Is Required for the Activation of Cardiac and Smooth Muscle Genes

Myocardin belongs to the SAF-A/B, Acinus, PIAS (SAP) domain family of transcription factors and is specifically expressed in cardiac and smooth muscle. Myocardin functions as a transcriptional coactivator of SRF and is sufficient and necessary for smooth muscle gene expression. We have previously found that myocardin induces the acetylation of nucleosomal histones surrounding SRF-binding sites in the control regions of cardiac and smooth muscle genes through recruiting chromatin-modifying enzyme p300, yet no studies have determined whether myocardin itself is similarly modified. In this study, we show that myocardin is a direct target for p300-mediated acetylation. p300 acetylates lysine residues at the N terminus of the myocardin protein. Interestingly, a direct interaction between p300 and myocardin, which is mediated by the C terminus of myocardin, is required for the acetylation event. Acetylation of myocardin by p300 enhances the association of myocardin and SRF as well as the formation of the myocardin-SRF-CArG box ternary complex. Conversely, acetylation of myocardin decreases the binding of histone deacetylase 5 (HDAC5) to myocardin. Acetylation of myocardin is required for myocardin to activate smooth muscle genes. Our study demonstrates that acetylation plays a key role in modulating myocardin function in controlling cardiac and smooth muscle gene expression

Novel Evolved Immunoglobulin (Ig)-Binding Molecules Enhance the Detection of IgM against Hepatitis C Virus

Detection of specific antibodies against hepatitis C virus (HCV) is the most widely available test for viral diagnosis and monitoring of HCV infections. However, narrowing the serologic window of anti-HCV detection by enhancing anti-HCV IgM detection has remained to be a problem. Herein, we used LD5, a novel evolved immunoglobulin-binding molecule (NEIBM) with a high affinity for IgM, to develop a new anti-HCV enzyme-linked immunosorbent assay (ELISA) using horseradish peroxidase-labeled LD5 (HRP-LD5) as the conjugated enzyme complex. The HRP-LD5 assay showed detection efficacy that is comparable with two kinds of domestic diagnostic kits and the Abbott 3.0 kit when tested against the national reference panel. Moreover, the HRP-LD5 assay showed a higher detection rate (55.9%, 95% confidence intervals (95% CI) 0.489, 0.629) than that of a domestic diagnostic ELISA kit (Chang Zheng) (53.3%, 95% CI 0.463, 0.603) in 195 hemodialysis patient serum samples. Five serum samples that were positive using the HRP-LD5 assay and negative with the conventional anti-HCV diagnostic ELISA kits were all positive for HCV RNA, and 4 of them had detectable antibodies when tested with the established anti-HCV IgM assay. An IgM confirmation study revealed the IgM reaction nature of these five serum samples. These results demonstrate that HRP-LD5 improved anti-HCV detection by enhancing the detection of anti-HCV IgM, which may have potential value for the early diagnosis and screening of hepatitis C and other infectious diseases

Abstracts from the 8th International Conference on cGMP Generators, Effectors and Therapeutic Implications

This work was supported by a restricted research grant of Bayer AG

Tetrandrine Inhibits Skeletal Muscle Differentiation by Blocking Autophagic Flux

Tetrandrine is well known to act as a calcium channel blocker. It is a potential candidate for a tumor chemotherapy drug without toxicity. Tetrandrine inhibits cancer cell proliferation and induces cell death through apoptosis and autophagy. As cancer patients usually experience complications with sarcopenia or muscle injury, we thus assessed the effects of tetrandrine on skeletal muscle cells. We report in this study that a low dose of tetrandrine (less than 5 μM) does not affect the proliferation of C2C12 myoblasts, but significantly inhibits myogenic differentiation. Consistently, tetrandrine inhibited muscle regeneration after BaCl2-induced injury. Mechanistic experiments showed that tetrandrine decreased the p-mTOR level and increased the levels of LC3 and SQSTM1/p62 during differentiation. Ad-mRFP-GFP-LC3B transfection experiments revealed that the lysosomal quenching of GFP signals was suppressed by tetrandrine. Furthermore, the levels of DNM1L/Drp1, PPARGA1 and cytochrome C (Cyto C), as well as caspase 3 activation and ROS production, were decreased following tetrandrine administration, indicating that the mitochondrial network signaling was inhibited. Our results indicate that tetrandrine has dual effects on autophagic flux in myoblasts during differentiation, activation in the early stage and blockade in the late stage. The ultimate blocking of autophagic flux by tetrandrine led to the disruption of mitochondria remodeling and inhibition of myogenic differentiation. The inhibitory effects of tetrandrine on skeletal muscle differentiation may limit its application in advanced cancer patients. Thus, great attention should be paid to the clinical use of tetrandrine for cancer therapy

Location decision of low-altitude service station for transfer flight based on modified immune algorithm

The location of Low-Altitude Flight Service Station (LAFSS) is a comprehensive decision work, and it is also a multi-objective optimization problem (MOOP) with constraints. As a swarm intelligence search algorithm for solving constrained MOOP, the Immune Algorithm (IA) retains the excellent characteristics of genetic algorithm. Using some characteristic information or knowledge of the problem selectively and purposefully, the degradation phenomenon in the optimization process can be suppressed and the global optimum can be achieved. However, due to the large range involved in the low-altitude transition flight, the geographical characteristics, economic level and service requirements among the candidate stations in the corridor are quite different, and the operational safety and service efficiency are interrelated and conflict with each other. And all objectives cannot be optimal. Therefore, this article proposes a Modified Immune Algorithm (MIA) with two-layer response to solve the constrained multi-objective location mathematical model of LAFSS. The first layer uses the demand track as the cell membrane positioning pattern recognition service response distance to trigger the innate immunity to achieve the basic requirements of security service coverage. In the second layer, the expansion and upgrading of adjacent candidate sites are compared to the pathogen’s effector, and the adaptive immunity is directly or indirectly triggered again through the cloning, mutation and reproduction between candidate sites to realize the multi-objective equilibrium of the scheme. Taking 486,000 km2 of Sichuan Province as an example, MIA for LAFSS is simulated by the MATLAB platform. Based on the Spring open source application framework of Java platform, the cesiumjs map data is called through easyui, and the visualization of site selection scheme is presented with the terrain data of Map World as the background. The experimental results show that, compared with dynamic programming and ordinary immunization, the immune trigger mode of double response and the improved algorithm of operation parameter combination designed by the Taguchi experiment, the total economic cost of location selection is reduced by 26.4%, the service response time is reduced by 25%, the repeat coverage rate is reduced by 29.5% and the effective service area is increased by 17.5%. The security risk, service efficiency and location cost are balanced. The present work is to provide an effective location method for the layout number and location of local transfer flight service stations. For complex scenes with larger scale of low-altitude flight supply and demand and larger terrain changes in the region, the above research methods can be used to effectively split and reduce the dimension

Fundamental study of cracking gasification process for comprehensive utilization of vacuum residue

The article is devoted to investigating some fundamentals about a residue cracking gasification (RCG) process for petroleum residues which integrates the catalytic cracking of the residue and the gasification of the cracking-generated coke. Three heat carrier particles were tested to show their different activities in cracking a vacuum residue (VR), finding that the synthesized kaolin catalyst allowed the higher liquid yield and higher conversion in comparison with silica sand and a commercial FCC catalyst. The parametric influences on the product distribution for VR cracking were thus studied over the kaolin catalyst for the major parameters including reaction temperature, catalyst-to-oil ratio, steam-to-oil ratio and different VRs. The conversion over 90% and the liquid yield above 80 wt.% were achieved at 500 degrees C under the optimized operating conditions. The VR with higher content of residual carbon was proven to generate more coke in the cracking. The coke deposited on the catalyst was well gasified via its interaction with steam and oxygen at 800 degrees C, and the CO and H-2 together was up to 80 vol.% in the produced syngas. The gasification-regenerated kaolin catalyst enabled the similar product distribution of VR cracking, thus justifying the technology feasibility for the tested RCG process. (C) 2013 Elsevier Ltd. All rights reserved

Identity-based provable data possession revisited: Security analysis and generic construction

2016 Elsevier B.V.Provable Data Possession (PDP), which enables cloud users to verify the data integrity without retrieving the entire file, is highly essential for cloud storage. Observing all the existing PDP schemes rely on the Public Key Infrastructure (PKI), Wang proposed an identity-based distributed provable data possession (ID-DPDP) scheme that can (1) eliminate the complex certificate management and (2) be applied to the multi-cloud scenario. The scheme is efficient, flexible and supports private verification, delegated verification and public verification. In this paper, we find that ID-DPDP is flawed since it fails to achieve soundness. We then fix the flaw by presenting a generic construction for identity-based PDP (ID-PDP) protocol, derived from secure digital signature schemes and traditional PDP protocols. We prove that the soundness of the generic ID-PDP construction depends on the security of the underlying PDP protocols and the signature schemes. An instance of the generic construction by utilizing a state-of-the-art PDP protocol due to Shacham and Waters and BLS short signature scheme is given. Moreover, a new ID-DPDP protocol is obtained by extending the basic ID-PDP to multiple clouds environment. The implementation shows that the proposed ID-PDP protocol is efficient