Quantitative and functional post-translational modification proteomics reveals that TREPH1 plays a role in plant thigmomorphogenesis

Plants can sense both intracellular and extracellular mechanical forces and can respond through morphological changes. The signaling components responsible for mechanotransduction of the touch response are largely unknown. Here, we performed a high-throughput SILIA (stable isotope labeling in Arabidopsis)-based quantitative phosphoproteomics analysis to profile changes in protein phosphorylation resulting from 40 seconds of force stimulation in Arabidopsis thaliana. Of the 24 touch-responsive phosphopeptides identified, many were derived from kinases, phosphatases, cytoskeleton proteins, membrane proteins and ion transporters. TOUCH-REGULATED PHOSPHOPROTEIN1 (TREPH1) and MAP KINASE KINASE 2 (MKK2) and/or MKK1 became rapidly phosphorylated in touch-stimulated plants. Both TREPH1 and MKK2 are required for touch-induced delayed flowering, a major component of thigmomorphogenesis. The treph1-1 and mkk2 mutants also exhibited defects in touch-inducible gene expression. A non-phosphorylatable site-specific isoform of TREPH1 (S625A) failed to restore touch-induced flowering delay of treph1-1, indicating the necessity of S625 for TREPH1 function and providing evidence consistent with the possible functional relevance of the touch-regulated TREPH1 phosphorylation. Bioinformatic analysis and biochemical subcellular fractionation of TREPH1 protein indicate that it is a soluble protein. Altogether, these findings identify new protein players in Arabidopsis thigmomorphogenesis regulation, suggesting that protein phosphorylation may play a critical role in plant force responses

Generation of diffuse large B cell lymphoma-associated antigen-specific Vα6/Vβ13+T cells by TCR gene transfer

<p>Abstract</p> <p>Background</p> <p>Our previous study had amplified antigen-specific full-length TCR α and β genes of clonally expanded T cells in the peripheral blood (PB) of patients with diffuse large B-cell lymphoma (DLBCL). The transfer of T cell receptor (TCR) genes endows T cells with new antigen specificity. Therefore, the aim of this study is to generate diffuse large B cell lymphoma (DLBCL)-specific T cells by T cell receptor (TCR) gene transfer.</p> <p>Materials and methods</p> <p>Two different eukaryotic expression plasmids harboring TCR Vα6 and TCR Vβ13 genes specific for DLBCL-associated antigens were constructed and subsequently transferred into human T cells using Nucleofector™ technique. The expression of targeted genes in TCR gene-modified cells was detected by real-time PCR, and western blot using TCR Vβ antibody. The specific cytotoxicity of TCR gene-transferred T cells <it>in vitro </it>was estimated using a lactate dehydrogenase (LDH) release assay.</p> <p>Results</p> <p>Two different eukaryotic expression plasmids harboring TCR Vα6 and TCR Vβ13 genes specific for DLBCL-associated antigens were constructed and subsequently transferred into T cells from healthy donors. Specific anti-DLBCL cytotoxic T lymphocytes (CTL) could be induced by transduction of specific TCR gene to modify healthy T cells. The transgene cassette of TCR Vβ13-IRES-TCR Vα6 was superior to the other in the function of TCR-redirected T cells.</p> <p>Conclusions</p> <p>Specific anti-DLBCL cytotoxic T lymphocyte (CTL) could be inducted by transduction of specific TCR gene to modify healthy T cells.</p

RCAgent: Cloud Root Cause Analysis by Autonomous Agents with Tool-Augmented Large Language Models

Large language model (LLM) applications in cloud root cause analysis (RCA) have been actively explored recently. However, current methods are still reliant on manual workflow settings and do not unleash LLMs' decision-making and environment interaction capabilities. We present RCAgent, a tool-augmented LLM autonomous agent framework for practical and privacy-aware industrial RCA usage. Running on an internally deployed model rather than GPT families, RCAgent is capable of free-form data collection and comprehensive analysis with tools. Our framework combines a variety of enhancements, including a unique Self-Consistency for action trajectories, and a suite of methods for context management, stabilization, and importing domain knowledge. Our experiments show RCAgent's evident and consistent superiority over ReAct across all aspects of RCA -- predicting root causes, solutions, evidence, and responsibilities -- and tasks covered or uncovered by current rules, as validated by both automated metrics and human evaluations. Furthermore, RCAgent has already been integrated into the diagnosis and issue discovery workflow of the Real-time Compute Platform for Apache Flink of Alibaba Cloud

A Data Transmission Algorithm Based on Dynamic Grid Division for Coal Goaf Temperature Monitoring

WSN (wireless sensor network) is a perfect tool of temperature monitoring in coal goaf. Based on the three-zone theory of goaf, the GtmWSN model is proposed, and its dynamic features are analyzed. Accordingly, a data transmission scheme, named DTDGD, is worked out. Firstly, sink nodes conduct dynamic grid division on the GtmWSN according to virtual semicircle. Secondly, each node will confirm to which grid it belongs based on grid number. Finally, data will be delivered to sink nodes with greedy forward and hole avoidance. Simulation results and field data showed that the GtmWSN and DTDGD satisfied the lifetime need of goaf temperature monitoring

Fuzzy sliding mode control of a multi-DOF parallel robot in rehabilitation environment

Multi-degrees of freedom (DOF) parallel robot, due to its compact structure and high operation accuracy, is a promising candidate for medical rehabilitation devices. However, its controllability relating to the nonlinear characteristics challenges its interaction with human subjects during the rehabilitation process. In this paper, we investigated the control of a parallel robot system using fuzzy sliding mode control (FSMC) for constructing a simple controller in practical rehabilitation, where a fuzzy logic system was used as the additional compensator to the sliding mode controller (SMC) for performance enhancement and chattering elimination. The system stability is guaranteed by the Lyapunov stability theorem. Experiments were conducted on a lower limb rehabilitation robot, which was built based on kinematics and dynamics analysis of the 6-DOF Stewart platform. The experimental results showed that the position tracking precision of the proposed FSMC is sufficient in practical applications, while the velocity chattering had been effectively reduced in comparison with the conventional FSMC with parameters tuned by fuzzy systems

Effect of Berberine on PPAR α

Rhizoma coptidis, the root of Coptis chinensis Franch, has been used in China as a folk medicine in the treatment of diabetes for thousands of years. Berberine, one of the active ingredients of Rhizoma coptidis, has been reported to improve symptoms of diabetes and to treat experimental cardiac hypertrophy, respectively. The objective of this study was to evaluate the potential effect of berberine on cardiomyocyte hypertrophy in diabetes and its possible influence on peroxisome proliferator-activated receptor-α (PPARα)/nitric oxide (NO) signaling pathway. The cardiomyocyte hypertrophy induced by high glucose (25.5 mmol/L) and insulin (0.1 μmol/L) (HGI) was characterized in rat primary cardiomyocyte by measuring the cell surface area, protein content, and atrial natriuretic factor mRNA expression level. Protein and mRNA expression were measured by western blot and real-time RT-PCR, respectively. The enzymatic activity of NO synthase (NOS) was measured using a spectrophotometric assay, and NO concentration was measured using the Griess assay. HGI significantly induced cardiomyocyte hypertrophy and decreased the expression of PPARα and endothelial NOS at the mRNA and protein levels, which occurred in parallel with declining NOS activity and NO concentration. The effect of HGI was inhibited by berberine (0.1 to 100 μmol/L), fenofibrate (0.3 μmol/L), or L-arginine (100 μmol/L). MK886 (0.3 μmol/L), a selective PPARα antagonist, could abolish the effects of berberine and fenofibrate. NG-nitro-L-arginine-methyl ester (100 μmol/L), a NOS inhibitor, could block the effects of L-arginine, but only partially blocked the effects of berberine. These results suggest that berberine can blunt HGI-induced cardiomyocyte hypertrophy in vitro, through the activation of the PPARα/NO signaling pathway

Simultaneous Extraction and Identification of Phenolic Compounds in Anoectochilus roxburghii Using Microwave-Assisted Extraction Combined with UPLC-Q-TOF-MS/MS and Their Antioxidant Activities

This study used MAE and RSM to extract phenolic compounds from Anoectochilus roxburghii, and the optimum conditions defined by the model to give an optimum yield of 1.31%. The antioxidant activity in vitro showed when the concentration of phenolic compounds was reached 1 mg mL-1, the clearance rates were 82.58% for DPPH and 97.62% for ABTS+. In vivo antioxidant experiments used D-galactose to build oxidative damage in healthy Kunming mice. The result showed that the extractions of A. roxburghii can improve the antioxidant ability and the medium and low dose groups had better ability to scavenge free radicals. The UPLC-Q-TOF-MS/MS was developed to identify 21 kinds of phenolic compounds by molecular mass, ms/ms fragmentation, as well as retention time. The result showed that the phenolic compounds of A. roxburghii had significant potential as a natural antioxidant to promote health and to reduce the risk of disease