pH-responsive bentonite nanoclay carriers control the release of benzothiazolinone to restrain bacterial wilt disease.

Ralstonia solanacearum (R. solanacearum) is one of the most devastating pathogens in terms of losses in agricultural production. Bentonite (Bent) is a promising synergistic agent used in development of effective and environmentally friendly pesticides against plant disease. However, the synergistic mechanism of Bent nanoclays with benzothiazolinone (BIT) against R. solanacearum is unknown. In this work, acid-functionalized porous Bent and cetyltrimethylammonium bromide (CTAB) were employed as the core nanoclays, and BIT was loaded into the clay to form BIT-loaded CT-Bent (BIT@CT-Bent) for the control of bacterial wilt disease. BIT@CT-Bent exhibited pH-responsive release behavior that fit the Fickian diffusion model, rapidly releasing BIT in an acidic environment (pH = 5.5). The antibacterial effect of BIT@CT-Bent was approximately 4 times greater than that of the commercial product BIT, and its biotoxicity was much lower than that of BIT under the same conditions. Interestingly, R. solanacearum attracted BIT@CT-Bent into the nanocomposites and induced cytoplasmic leakage and changes in membrane permeability, indicating an efficient and synergistic bactericidal effect that rapidly reduced bacterial density. In addition, BIT@CT-Bent significantly inhibited R. solanacearum biofilm formation and swimming activity, by suppressing the expression of phcA, solR and vsrC. Indeed, exogenous application of BIT@CT-Bent significantly suppressed the virulence of R. solanacearum on tobacco plants, with control effect of 75.48%, 72.08% and 66.08% at 9, 11 and 13 days after inoculation, respectively. This study highlights the potential of using BIT@CT-Bent as an effective, eco-friendly bactericide to control bacterial wilt diseases and for the development of sustainable crop protection strategies.

Molecular mechanism of plant elicitor daphnetin-carboxymethyl chitosan nanoparticles against Ralstonia solanacearum by activating plant system resistance.

The exploration of biopolymer-based materials to avoid hazardous chemicals in agriculture has gained enormous importance for sustainable crop protection. Due to its good biocompatibility and water solubility, carboxymethyl chitosan (CMCS) has been widely applied as a pesticide carrier biomaterial. However, the mechanism by which carboxymethyl chitosan-grafted natural product nanoparticles induce tobacco systemic resistance against bacterial wilt remains largely unknown. In this study, water-soluble CMCS-grafted daphnetin (DA) nanoparticles (DA@CMCS-NPs) were successfully synthesized, characterized, and assessed for the first time. The grafting rate of DA in CMCS was 10.05 %, and the water solubility was increased. In addition, DA@CMCS-NPs significantly increased the activities of CAT, PPO and SOD defense enzymes, activated the expression of PR1 and NPR1, and suppressed the expression of JAZ3. DA@CMCS-NPs could induce immune responses against R. solanacearum in tobacco, including increases in defense enzymes and overexpression of pathogenesis-related (PR) proteins. The application of DA@CMCS-NPs effectively suppressed the development of tobacco bacterial wilt in pot experiments, and the control efficiency was as high as 74.23 %, 67.80 %, 61.67 % at 8, 10, and 12 days after inoculation. Additionally, DA@CMCS-NPs has excellent biosafety. Therefore, this study highlighted the application of DA@CMCS-NPs in manipulating tobacco to generate defense responses against R. solanacearum, which can be attributed to systemic resistance.

Comprehensive structural characterization of the human AAA+ disaggregase CLPB in the apo- and substrate-bound states reveals a unique mode of action driven by oligomerization.

The human AAA+ ATPase CLPB (SKD3) is a protein disaggregase in the mitochondrial intermembrane space (IMS) and functions to promote the solubilization of various mitochondrial proteins. Loss-of-function CLPB mutations are associated with a few human diseases with neutropenia and neurological disorders. Unlike canonical AAA+ proteins, CLPB contains a unique ankyrin repeat domain (ANK) at its N-terminus. How CLPB functions as a disaggregase and the role of its ANK domain are currently unclear. Herein, we report a comprehensive structural characterization of human CLPB in both the apo- and substrate-bound states. CLPB assembles into homo-tetradecamers in apo-state and is remodeled into homo-dodecamers upon substrate binding. Conserved pore-loops (PLs) on the ATPase domains form a spiral staircase to grip and translocate the substrate in a step-size of 2 amino acid residues. The ANK domain is not only responsible for maintaining the higher-order assembly but also essential for the disaggregase activity. Interactome analysis suggests that the ANK domain may directly interact with a variety of mitochondrial substrates. These results reveal unique properties of CLPB as a general disaggregase in mitochondria and highlight its potential as a target for the treatment of various mitochondria-related diseases.

PPICT: an integrated deep neural network for predicting inter-protein PTM cross-talk.

Post-translational modifications (PTMs) fine-tune various signaling pathways not only by the modification of a single residue, but also by the interplay of different modifications on residue pairs within or between proteins, defined as PTM cross-talk. As a challenging question, less attention has been given to PTM dynamics underlying cross-talk residue pairs and structural information underlying protein-protein interaction (PPI) graph, limiting the progress in this PTM functional research. Here we propose a novel integrated deep neural network PPICT (Predictor for PTM Inter-protein Cross-Talk), which predicts PTM cross-talk by combining protein sequence-structure-dynamics information and structural information for PPI graph. We find that cross-talk events preferentially occur among residues with high co-evolution and high potential in allosteric regulation. To make full use of the complex associations between protein evolutionary and biophysical features, and protein pair features, a heterogeneous feature combination net is introduced in the final prediction of PPICT. The comprehensive test results show that the proposed PPICT method significantly improves the prediction performance with an AUC value of 0.869, outperforming the existing state-of-the-art methods. Additionally, the PPICT method can capture the potential PTM cross-talks involved in the functional regulatory PTMs on modifying enzymes and their catalyzed PTM substrates. Therefore, PPICT represents an effective tool for identifying PTM cross-talk between proteins at the proteome level and highlights the hints for cross-talk between different signal pathways introduced by PTMs.

Deglacial Subantarctic CO2 outgassing driven by a weakened solubility pump.

The Subantarctic Southern Ocean has long been thought to be an important contributor to increases in atmospheric carbon dioxide partial pressure (pCO2) during glacial-interglacial transitions. Extensive studies suggest that a weakened biological pump, a process associated with nutrient utilization efficiency, drove up surface-water pCO2 in this region during deglaciations. By contrast, regional influences of the solubility pump, a process mainly linked to temperature variations, have been largely overlooked. Here, we evaluate relative roles of the biological and solubility pumps in determining surface-water pCO2 variabilities in the Subantarctic Southern Ocean during the last deglaciation, based on paired reconstructions of surface-water pCO2, temperature, and nutrient utilization efficiency. We show that compared to the biological pump, the solubility pump imposed a strong impact on deglacial Subantarctic surface-water pCO2 variabilities. Our findings therefore reveal a previously underappreciated role of the solubility pump in modulating deglacial Subantarctic CO2 release and possibly past atmospheric pCO2 fluctuations.

[Correlation of gut microbiota and ischemic stroke: a review].

With the widespread application of next-generation sequencing(NGS), especially 16 S rRNA and shotgun sequencing, researchers are no longer troubled with massive data on the gut microbiota, and the correlation between the gut microbiota and the brain(central nervous system) has been gradually revealed. Research on the microbiota-gut-brain axis(MGBA) based on the gut microbiota have provided insights into the exploration of the pathogenesis and risk factors of ischemic stroke(IS), a cerebrovascular disease with high disability and mortality rates, and also facilitate the selection of therapeutic targets of this class of drugs. This study reviewed the application of NGS in the study of gut microbiota and the research progress of MGBA in recent years and systematically collated the research papers on the correlation between IS and gut microbiota. Furthermore, from the bi-directional regulation of MGBA, this study also discussed the high-risk factors of IS under the dysregulation of gut microbiota and the pathophysiological changes of gut microbiota after the occurrence of IS and summarized the related targets to provide a reliable reference for the therapeutic research of IS from the gut microbiota.

In Situ Programmable DNA Circuit-Promoted Electrochemical Characterization of Stemlike Phenotype in Breast Cancer.

Breast cancer is one of the most common malignant diseases among women worldwide, and the existence of breast cancer stem cells is closely associated with poor outcomes. Herein, we report an electrochemical phenotyping method to characterize the stemlike phenotype in breast cancer, offering a low-cost but robust choice other than the highly expensive and experience-dependent flow cytometry. Specially, after immune-magnetic beads-assisted enrichment, an in situ programmable DNA circuit is designed using capture probes to bring in the toeholds for DNA assembly and effector probes to accelerate the removal of background signals. The electrochemical phenotyping method could sensitively determine breast cancer stem cells in a wide linear range and exhibit desirable accuracy and reliability. The method can not only monitor the phenotypic transition of breast cancer cells and the drug-reversed effect but also determinate stemlike phenotype in the mice bearing breast cancer xenograft tumor. Overall, the electrochemical phenotyping method may provide promising technical support for precise management of breast tumors.

Kaempferol Reverses Aerobic Glycolysis via miR-339-5p-Mediated PKM Alternative Splicing in Colon Cancer Cells.

Colon cancer is an aggressive malignancy with very limited therapeutic approaches. The available therapeutic agents for colon cancer show strong adverse effects and poor effectiveness, indicating the urgent need to identify new therapeutic drugs for this malignancy. Kaempferol, a flavonoid found in a variety of natural foods, exhibits significant inhibitory effects on colon cancer. Here, it was found that kaempferol inhibited the proliferation of human colon cancer cells HCT116 and DLD1 in a dose-dependent manner, and the IC50 values were 63.0 ± 12.9 and 98.3 ± 15.9 µM, respectively. Also, kaempferol treatment delayed G1 phase progression of cell cycle and induced apoptosis. Aerobic glycolysis is the major energy source for various tumor growths, including colon cancer. Indeed, kaempferol treatment impaired glucose consumption, which subsequently led to reduced lactic acid accumulation and ATP production. Mechanistically, kaempferol promoted the expression of miR-339-5p. Further studies identified hnRNPA1 and PTBP1 as two direct targets of miR-339-5p. By directly targeting hnRNPA1 and PTBP1, miR-339-5p reduced the expression of M2-type pyruvate kinase (PKM2) but induced that of PKM1. In conclusion, these data demonstrate that by modulating miR-339-5p-hnRNPA1/PTBP1-PKM2 axis, kaempferol inhibits glycolysis and colon cancer growth, which reveals a new explanation for the molecular mechanism underlying kaempferol anti-tumor.

A catalytic molecule machine-driven biosensing method for amplified electrochemical detection of exosomes.

Nowadays, exosomes that carry abundant information have attracted increasing attention as potent biomarkers of liquid biopsy and ideal candidates for early diagnosis and treatment of cancers. In this work, we propose a "principle-of-proof" biosensing method for amplified electrochemical detection of exosomes by using HepG2-derived exosomes as models. Specifically, target exosomes are enriched on anti-CD63-functionalized immunobeads and then recognized by a DNA chain containing CD63 aptamer region, which subsequently initiates a catalytic molecule machine that relies on cascade toehold-mediated strand displacement reaction. Benefiting from high efficiency of the molecule machine, the method shows a linear range from 1 × 105 to 5 × 107 particles/mL and a detection limit of 1.72 × 104 particles/mL toward target exosomes, better than most existing detection methods. Moreover, the method demonstrates a high specificity even in serum samples and suggests a potential use in clinic, which may provide sufficient information for disease diagnosis, especially early detection and prognosis monitoring of tumors.

Self-Assembling Peptide-Based Multifunctional Nanofibers for Electrochemical Identification of Breast Cancer Stem-like Cells.

Cancer stem-like cells are closely related with the development and metastasis of tumors. Herein, an electrochemical method is proposed to identify stem-like cells in breast tumor. The core concept of the method is the use of multifunctional nanofibers (MNFs), which are synthesized through facile self-assembly of peptide probes. MNFs can perform three functions, specifically targeting surface biomarker to identify stem-like cells, recruiting silver nanoparticles (AgNPs) to generate electrochemical signals, and providing large amounts of reaction sites to amplify signals. Specially, breast cancer stem cells (BCSCs) are first captured by nucleolin aptamer immobilized on the electrode surface and then selectively recognized by MNFs through the binding with CD44, thereby offering a large number of azide groups for signal labeling. By tracing electrochemical signals from MNF-recruited AgNPs, the method demonstrates to detect target cells as low as 6 cells/mL within a wide linear range from 10 to 5 × 105 cells/mL. Moreover, the method can not only recognize BCSCs with high selectivity in complex environment but also monitor drug-induced stemness changes with high sensitivity, providing promising prospective clinic applications in the future.

Cellular interface supported toehold strand displacement cascade for amplified dual-electrochemical signal and its application for tumor cell analysis.

In this work, toehold strand displacement cascade (TSDC) has been delicately designed and carried out on the cellular interface for the amplification and output of dual-electrochemical signal. Specifically, antibody cross-linked T strand can recognize cell which is linked with immune-magnetic bead. Subsequently, T strand on the cellular interface can mediate the occurrence of TSDC, resulting the change of SN3/S1-MB to SN3/S2-Fc ratio in the supernatant after magnetic separation. The resultant SN3/S1-MB and SN3/S2-Fc can be immobilized on the electrode interface through click chemistry and give the amplified double electrochemical signal. So the tumor cell amount can closely correlate with the change of the double signal. Except for output of the double signal for improvement of analytical accuracy, the double magnetic separation not only eliminate the interference of the complicated substances in serum, but also remove the influence of cell on click reaction on the electrode interface. So based on cellular interface supported TSDC for amplified dual-electrochemical signal, the established method has been successfully applied to analyze the tumor cells in serum accurately and sensitively.

Integration of fluorescence imaging and electrochemical biosensing for both qualitative location and quantitative detection of cancer cells.

In this work, DNA-templated silver nanoclusters (DNA-AgNCs) with unique fluorescent and electrochemical properties are prepared as dual signal probes for both qualitative imaging and quantitative detection of cancer cells in an integrated system. ITO electrode that has good light transmittance and electric conductivity is employed as a substrate for dual analysis of cancer cells. ITO electrode is firstly modified by AS1141 aptamer, which could selectively bind to nucleolin overexpressed on the surface of a model breast cancer cell, MCF-7 cell line. The composite of mucin 1 antibody (anti-MUC1) and DNA-AgNCs then binds to MUC1 on the surface of captured MCF-7 cell, forming a sandwich-like structure. Therefore, our method allows noninvasive fluorescence imaging and amplified electrochemical detection using a single labeling platform, providing a biocompatible and highly specific method for adequate analysis of cancer cells. Experimental results demonstrate that strong red fluorescence of DNA-AgNCs clearly displays the loading of cancer cells on ITO electrode after dual recognition, and amplified electrochemical signals of DNA-AgNCs enable improved sensitivity toward quantitative analysis with a detection limit of 3 cells.

Peptide-templated multifunctional nanoprobe for feasible electrochemical assay of intracellular kinase.

Protein kinases play a critical role in regulation of intracellular signal transduction, whose aberrant expression is closely associated with various dangerous human diseases. In this paper, we propose a feasible electrochemical assay of intracellular kinase by incorporating peptide nanoprobe-assisted signal labeling and signal amplification. Protein kinase A (PKA)-specific peptide P1 is self-assembled on the surface of a gold electrode, serine of which could be phosphorylated with catalysis of PKA in the presence of adenosine-5'-triphosphate (ATP). Another artificial peptide P2 contains a short template for preparation of copper nanoparticles-based nanoprobe (P2-CuNPs) and provides arginine residues for specific recognition of phosphorylation site. After PKA-catalyzed phosphorylation, phosphorylated P1 specially binds with P2-CuNPs through ultra-stable phosphate-guanidine interaction, and thus results in amplified electrochemical response from surface-attached CuNPs. Our method demonstrates a satisfactory sensitivity toward PKA detection with a detection limit of 0.0019 U/mL, which is also successfully applied in intracellular PKA assay and inhibitory study with high specificity comparable to ELISA. Therefore, the facile method suggests a promising potential use in kinase-related biochemical fundamental research, disease diagnosis and drug discovery in the future.