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When a multilayer ceramic capacitor (MLCC) is soldered on a printed circuit board (PCB), the capacitor deformation generates a force on the PCB, which serves as a source to excite PCB vibration and generate unwanted acoustic noise. A MLCC can be used under any loading conditions as long as the power supply voltage remains below its rated voltage; thus, the MLCC source mode can be extremely complicated because of different combinations including direct current voltage levels, alternating current noise amplitudes, and related frequencies. When the power rail is under different loading conditions, the PCB vibration amplitude can differ; however, the MLCC vibration source model should remain the same under a given condition. In this article, the MLCC equivalent source models are developed with a simplified method, and a source model library for the six MLCCs is constructed. A method for simplifying the MLCC equivalent source model is addressed from three aspects: frequency, power supply dc voltage, and ac noise amplitude. The developed source model library is validated by comparing the simulated and measured system vibration velocity. The proposed simplification method can significantly reduce model development efforts and enable the creation of a comprehensive source model library for MLCCs.

Multilayer ceramic capacitors (MLCCs) are commonly used on printed circuit board (PCB) power distribution networks (PDN) for decoupling purposes. However, an MLCC can act as a source to excite PCB vibrations due to noise ripples on the power rail. In this letter, four types of equivalent source configurations are studied and compared to represent the excitation of an MLCC on a PCB driven by power rail noise. The corresponding source values are extracted at test-vehicle PCB resonance frequencies. The investigation of the four equivalent source types involves a comparison between simulated and measured PCB vibration velocity responses triggered by MLCC deformation across an audible frequency band. The advantages of each equivalent source are discussed for specific application scenarios.

Four Ag(I) complexes with mefenamato and nitrogen heterocyclic ligands, [Ag(2-apy)(mef)]2(1), [Ag(3-apy)(mef)] (2), [Ag2(tmpyz)(mef)2] (3), and {[Ag(4,4′-bipy)(mef)]2(CH3CN)1.5(H2O)2}n(4), (mef = mefenamato, 2-apy = 2-aminopyridine, 3-apy = 3-aminopyridine, tmpyz = 2,3,5,6-tetramethylpyrazine, 4,4′-bipy = 4,4′-bipyridine), were synthesized and characterized. The interactions of these complexes with BSA were investigated by fluorescence spectroscopy, which indicated that these complexes quench the fluorescence of BSA by a static mechanism. The fluorescence data also indicated that the complexes showed good affinity for BSA, and one binding site on BSA was suitable for the complexes. The in vitrocytotoxicity of the four complexes against human cancer cell lines (MCF-7, HepG-2, A549, and MDA-MB-468) and one normal cell line (HTR-8) was evaluated by the MTT assay. Complex 1displayed high cytotoxic activity against A549 cells. Further studies revealed that complex 1could enhance the intracellular levels of ROS (reactive oxygen species) in A549 cells, cause cell cycle arrest in the G0/G1 phase, and induce apoptosis in A549 cells in a dose-dependent manner.

Exosomes (EXOs) are a class of natural nanovesicles secreted endogenously by mammalian cells. Recent decades have witnessed the broad applications of exosomes in cancer diagnosis and treatment. In the present study, an exosome-based drug delivery system employing synergistic miRNA-based autophagy inhibition and 7-coumarin-based chemotherapy was established for ovarian cancer therapy. Taking advantage of the outstanding biocompatibility and immune evasion, exosomes are used as carriers for targeted drug delivery. Magnetic nanoparticles (MNPs) were coupled to exosomes to improve the separation efficiency of exosomes. Two drugs, miRNA-FAM and 7-coumarin, were loaded into EXOs-MNPs by electroporation to achieve the miRNA-FAM/7-coumarin@EXOs-MNPs drug delivery system. The engineered exosome-based drug delivery system improved the cell entry ability of drugs and the targeting of cancer cells. Simultaneous application of the two drugs significantly improved the efficacy of cancer cell therapy, indicating that the two drugs have synergistic effects. Therefore, this work provided a new drug delivery system and synergistic therapy idea, which showed great potential for future clinical applications.

On-board electrical components can cause printed circuit board (PCB) vibration, thus generating audio noise if the electrical noise is in the audible frequency range. The electrical component-induced vibration can be equated to an external force applied to the PCB. This article presents a novel methodology to extract the equivalent force of electrical components on a PCB to study board vibration and potential acoustic noise problems. The method is based on a combination of measurement and simulation, wherein PCB vibration is used as the medium in the extraction process. The methodology is validated by the correlation of PCB vibration pattern, frequency, and amplitude with a known electromagnetic force applied to the PCB.

We mainly study an approximation theorem and generic uniqueness of weakly Pareto-Nash equilibrium (PNE) for multiobjective population games. First, we define the concept of approximate weakly PNE of multiobjective population games. Then, an approximation theorem is proved under very mild conditions. Furthermore, we prove the generic uniqueness of weakly PNE for multiobjective population games by establishing a function space. Finally, we obtain the generic uniqueness of weakly PNE for multiobjective population games in the case of function perturbation. The above results are new and improve the related results of the recent literature.

Heart failure with preserved ejection fraction (HFpEF) is a complex clinical syndrome with cardiac dysfunction, fluid retention and reduced exercise tolerance as the main manifestations. Current treatment of HFpEF is using combined medications of related comorbidities, there is an urgent need for a modest drug to treat HFpEF. Geniposide (GE), an iridoid glycoside extracted from Gardenia Jasminoides, has shown significant efficacy in the treatment of cardiovascular, digestive and central nervous system disorders. In this study we investigated the therapeutic effects of GE on HFpEF experimental models in vivo and in vitro. HFpEF was induced in mice by feeding with HFD and L-NAME (0.5 g/L) in drinking water for 8 weeks, meanwhile the mice were treated with GE (25, 50 mg/kg) every other day. Cardiac echocardiography and exhaustive exercise were performed, blood pressure was measured at the end of treatment, and heart tissue specimens were collected after the mice were euthanized. We showed that GE administration significantly ameliorated cardiac oxidative stress, inflammation, apoptosis, fibrosis and metabolic disturbances in the hearts of HFpEF mice. We demonstrated that GE promoted the transcriptional activation of Nrf2 by targeting MMP2 to affect upstream SIRT1 and downstream GSK3β, which in turn alleviated the oxidative stress in the hearts of HFpEF mice. In H9c2 cells and HL-1 cells, we showed that treatment with GE (1 μM) significantly alleviated H2O2-induced oxidative stress through the MMP2/SIRT1/GSK3β pathway. In summary, GE regulates cardiac oxidative stress via MMP2/SIRT1/GSK3β pathway and reduces cardiac inflammation, apoptosis, fibrosis and metabolic disorders as well as cardiac dysfunction in HFpEF.

Epigenetic regulation plays an important role in regulating the transgenerational toxicity of pollutants. However, the underlying mechanism of microRNAs (miRNAs) in regulating transgenerational nanoplastic toxicity remains largely unclear. We aimed to determine the miRNA-mediated mechanism for the induction of transgenerational nanoplastic toxicity. In Caenorhabditis elegans, although germline RNAi of both mir-240and mir-36suppressed polystyrene nanoparticle (PS-NP) toxicity, exposure to PS-NPs (1–100 μg L−1) only increased mir-240expression. A transgenerational increase in mir-240expression was observed after PS-NP exposure at P0 generation (P0-G), and the germline RNAi of mir-240suppressed transgenerational PS-NP toxicity. Among the predicted target genes of mir-240in the germline, the exposure to PS-NPs (1–100 μg L−1) decreased rab-5and rab-6.2expressions, whereas the germline RNAi of mir-240only increased rab-5expression in PS-NP exposed nematodes. A transgenerational decrease in rab-5expression was detected after PS-NP exposure at P0-G, and the germline RNAi of rab-5strengthened transgenerational PS-NP toxicity. Moreover, the resistance of mir-240(RNAi)to transgenerational PS-NP toxicity in inhibiting locomotion behavior and in reducing the brood size was inhibited by the germline RNAi of rab-5. Among the secreted ligands, the germline RNAi of rab-5increased the expressions of genes encoding insulin peptides (ins-3, ins-39, and daf-28), FGF ligand (egl-17), and ephrin ligand (efn-3) in PS-NP exposed nematodes and their corresponding receptor genes (daf-2, egl-15, and vab-1) in the offspring of PS-NP exposed nematodes. Therefore, an increase in germline mir-240mediated transgenerational PS-NP toxicity through insulin, FGF, and ephrin signals by affecting its target RAB-5. Our data demonstrated the important involvement of germline microRNA in mediating nanoplastic toxicity across multiple generations in organisms.

In this paper, a label-free electrochemical immunosensor for sensitive detection of prostate antigen (PSA) was developed based on a NiFe PBA/AuNPs composite. The prostate antigen antibody was immobilized and the immunosensor was constructed by using a glassy carbon electrode modified with a nanocomposite consisting of nickel–iron Prussian blue analog (NiFe PBA) and gold nanoparticles (AuNPs). Due to the good biological affinity of AuNPs for biomolecules, as well as the porous nanostructure and regular shape of NiFe PBA, NiFe PBA/AuNPs nanocomposites significantly improve the electron transport rate, while achieving excellent performance for the sensor. Due to the interaction between the antibody and the antigen on the modified electrode, the current signal of the NiFe PBA itself is reduced due to the redox changes in Fe2+and Fe3+, which can be determined by differential pulse voltammetry (DPV). Therefore, the monitoring of prostate antigen detection is realized. Under optimal experimental conditions, the immunosensor exhibited excellent detection performance with a dynamic response range from 0.5 pg mL−1to 1000 pg mL−1for the PSA concentration and a detection limit of 0.23 pg mL−1(S/N = 3). In addition, the PSA aptasensor has good selectivity, high stability, and satisfactory reproducibility and has broad potential in clinical research and diagnostic applications.

N-(1,3-Dimethylbutyl)-N′-phenyl-p-phenylenediamine (6-PPD), one of the most common additives used in rubber, enters the environment due to significant emissions of tire wear particles. 6-PPD quinone (6-PPDQ) is an important derivative of 6-PPD after ozonization. With concentrations ranging from nanograms per liter to μg/L, 6-PPDQ has so far been identified in a series of water samples. Acute lethality of 6-PPDQ in coho salmon (LC50< 1 μg/L) was lower than environmental concentrations of 6-PPDQ, highlighting the environment exposure risks of 6-PPDQ. It is becoming increasingly necessary to investigate the potential toxicity of 6-PPDQ at environmental concentrations. Here, we examined the effect of 6-PPDQ exposure on lifespan and healthspan and the underlying mechanism in Caenorhabditis elegans. Exposure to 6-PPDQ (1 and 10 μg/L) shortened the lifespan. Meanwhile, during the aging process, 6-PPDQ (0.1–10 μg/L) could decrease both pumping rate and locomotion behavior, suggesting the 6-PPDQ toxicity on healthspan. For the underlying molecular mechanism, the dysregulation in the insulin signaling pathway was linked to toxicity of 6-PPDQ on lifespan and healthspan. In the insulin signaling pathway, DAF-2 restricted the function of DAF-16 to activate downstream targets (SOD-3 and HSP-6), which in turn controlled the toxicity of 6-PPDQ on lifespan and healthspan. Additionally, in response to 6-PPDQ toxicity, insulin peptides (INS-6, INS-7, and DAF-28) could activate the corresponding receptor DAF-2. Therefore, exposure to 6-PPDQ at environmentally relevant concentrations potentially causes damage to both lifespan and healthspan by activating insulin signaling in organisms.

Acute myocardial infarction (AMI) is the most important cause of death among cardiovascular diseases. Long noncoding RNAs (lncRNAs) have been widely implicated in the regulation of AMI progression. Discrimination antagonizing nonprotein coding RNA (DANCR) alleviated hypoxia‐caused cardiomyocyte damages, and the underlying mechanisms remain unclear. Here, we investigated the function and mechanism of DANCR in hypoxia‐induced cardiomyocytes and AMI model by enzyme‐linked immunosorbent assay, reactive oxygen species and adenosine triphosphate measurement, and mitochondrial activity determination. Additionally, luciferase reporter assay, immunoblotting, and qRT‐PCR were performed to validate the interactions between DANCR/miR‐509‐5p and miR‐509‐5p/Kruppel‐like factor 13 (KLF13). The role of DANCR was also verified in AMI model by overexpression. Our results showed that DANCR expression was significantly downregulated in hypoxia‐induced cardiomyocytes or AMI model. Overexpression of DANCR significantly alleviated mitochondrial damages, reduced inflammation, and improved cardiac function in the AMI model. Furthermore, we demonstrated that miR‐509‐5p/KLF13 axis mediated the protective effect of DANCR. The current study highlighted the critical role of DANCR in alleviating AMI progression through targeting the miR‐509‐5p/KLF13 signaling axis, suggesting that DANCR may serve as a potential diagnostic marker or therapeutic target for AMI.

Term extraction aims to extract domain-specific terminology from documents. Secondary school mathematics terminology plays a guiding role in the learning and teaching of secondary school mathematics. We found that there needs to be more work in this field and that there are two main issues in extracting the terms from the middle school mathematics text: the polysemy of one character problem and nested terms problem. Accordingly, we have constructed a secondary school mathematics corpus and proposed a model to resolve these problems, termed BERT-LLA-CRF. Using pretraining to obtain dynamic word vectors to improve the model’s understanding of text semantics and solve the problem of multiple meanings of one word, the proposed model integrates attention mechanisms and Chinese character information to solve the nested terminology problem. The experiment results show that the proposed model effectively solves those problems and achieves good middle-school mathematics term extraction results.

Magnetic refrigeration technology based on the magnetocaloric effect can better meet the requirements of efficient energy consumption and conversion. From engineering perspective, preparing magnetocaloric composites is an effective and efficient approach to combine desirable magnetocaloric performance, large thermal conductivity and ameliorated mechanical properties. In the present work, Ni-Mn-Ga/Cu magnetocaloric composites were prepared by spark plasma sintering (SPS) and compared with their counterparts fabricated viaconventional methods. Magnetic properties were studied in detail and the magnetocaloric effect of Ni-Mn-Ga/Cu composites was characterized to be better than that of their hot-pressed counterparts and that of many micro/nano-sized Ni-Mn-Ga alloys. Besides, the composites exhibited favorable thermal conductivity of 11.2 W mK−1. The Hassel-man-Johnson model was adopted and modified to relate the thermal conductivity to the microstructure of the composites. Compared with arc-melted alloys, these SPS magnetocaloric composites sintered at different temperatures exhibited largely enhanced mechanical performance with minimum fracture stress of 340 MPa and strain of 4%. Furthermore, the failure mechanism of Ni-Mn-Ga/Cu magnetocaloric composite was elucidated by finite element simulation based on an extended linear Drucker-Prager model. The above findings from both experiments and simulation advance the knowledge of magnetocaloric materials and promote the development of magnetic refrigeration technology towards practical applications.

In this paper, we propose a novel four-stage data augmentation approach to ResNet-Conformer based acoustic modeling for sound event localization and detection (SELD). First, we explore two spatial augmentation techniques, namely audio channel swapping (ACS) and multi-channel simulation (MCS), to deal with data sparsity in SELD. ACS and MDS focus on augmenting the limited training data with expanding direction of arrival (DOA) representations such that the acoustic models trained with the augmented data are robust to localization variations of acoustic sources. Next, time-domain mixing (TDM) and time-frequency masking (TFM) are also investigated to deal with overlapping sound events and data diversity. Finally, ACS, MCS, TDM and TFM are combined in a step-by-step manner to form an effective four-stage data augmentation scheme. Tested on the Detection and Classification of Acoustic Scenes and Events (DCASE) 2020 data set, our proposed augmentation approach greatly improves the system performance, ranking our submitted system in the first place in the SELD task of the DCASE 2020 Challenge. Furthermore, we employ a ResNet-Conformer architecture to model both global and local context dependencies of an audio sequence and win the first place in the DCASE 2022 SELD evaluations.

To design a promising Al—Si filler alloy with a relatively low melting-point, good strength and plasticity for the Cu/Al joint, the Cu, Ni, Zr and Er elements were innovatively added to modify the traditional Al—Si eutectic filler. The microstructure and mechanical properties of filler alloys and Cu/Al joints were investigated. The result indicated that the Al—Si—Ni—Cu filler alloys mainly consisted of Al(s,s), Al2(Cu,Ni) and Si(s,s). The Al—10Si—2Ni—6Cu filler alloy exhibited relatively low solidus (521 °C) and liquidus (577 °C) temperature, good tensile strength (305.8 MPa) and fracture elongation (8.5%). The corresponding Cu/Al joint brazed using Al—10Si—2Ni—6Cu filler was mainly composed of Al8(Mn,Fe)2Si, Al2(Cu,Ni)3, Al(Cu,Ni), Al2(Cu,Ni) and Al(s,s), yielding a shear strength of (90.3±10.7) MPa. The joint strength was further improved to (94.6±2.5) MPa when the joint was brazed using the Al—10Si—2Ni—6Cu—0.2Er—0.2Zr filler alloy. Consequently, the (Cu, Ni, Zr, Er)-modified Al—Si filler alloy was suitable for obtaining high-quality Cu/Al brazed joints.

Recent studies on autophagy demonstrated a new extracellular secretion pathway for autophagosomes in addition to the routinely described intracellular degradation pathway. Besides, the secretory autophagosomes were found closely related to the occurrence and development of cancers. Therefore, analysis of the protein expression on secretory autophagosomes is a promising noninvasive strategy for cancer diagnosis and mechanism study. Herein, we constructed a three-dimensional (3D) microfluidic chip employing a fusiform micropillar array and layer-by-layer modification of gelatins, which obviously enhanced the mass transfer between reactants and increased the immobilization sites for capture antibody. As a result, the autophagosome capture efficiency of the 3D chip (74%) is significantly higher than that of the unmodified flat chip (47%). Using a two-step immunoreaction, ovarian cancer cell-secreted autophagosomes were successfully captured and detected. The results showed that two proteins, LC3B and HSP60 at the surface of autophagosomes, can be detected with limits of detection (LODs) of 141 particles μL–1and 126 particles μL–1, respectively. In addition, both LC3B and HSP60 expressions on autophagosomes can be used to distinguish the serum samples between cancer patients and healthy people, with a pvalue less than 0.01 (statistically significant difference) or 0.05 (statistically different), respectively. Moreover, the summed signal of LC3B and HSP60 showed a pvalue less than 0.001 (extremely statistically significant difference), demonstrating the good potential of this chip for further application in cancer diagnosis.

Graphical abstract:

This study presents a novel series of structural color coatings synthesized by blending fluorinated acrylate emulsion (FPA) with light absorbers (T6721) within polystyrene (PS) microspheres. The innovative strategy of replacing inter-microspherical air with FPA, a material of higher refractive index, has significantly boosted the reflectivity of the colloidal crystals to 95.86 %, particularly enhancing UVA reflectivity to a peak of 89 %. These coatings also showcase exceptional UV stability, structural integrity, and improved hydrophobicity, resisting UV-induced fading. The coatings' performance profiles recommend their use in demanding applications such as the coatings and painting industries, with potential for broader adoption in high-performance sectors including aerospace and automotive. This research advances the field of structural color technology, offering a sustainable solution for durable and vibrant pigment applications. Future efforts will aim to scale the synthesis and expand industrial applications.

Guided waves can propagate from flat plates to curved structures and carry structural health information. In this paper, an ultrasonic guided wave inspection method is proposed for detecting damages in curved parts made of carbon fibre reinforced plastics (CFRPs), such as hat-stringer-stiffened panel. By collecting full wavefield information, we obtain the standing wave information formed by the interaction between guided waves and damage. The interaction mechanism between guided waves and damage is simulated through finite element simulation, and the wavefield curvature imaging algorithm is used to generate a damage image in the curved area. Furthermore, a specimen with delamination damages is fabricated, and full wavefield information is collected through a scanning laser Doppler vibrometer (SLDV). We then use the proposed imaging algorithm to detect delamination damages of different sizes. The experimental results were consistent with the simulation results, confirming the effectiveness of the proposed method. Notably, the technology combines the advantages of guided waves and SLDV, which can obtain full wavefield information without contact, including damage-related characteristics. This work provides a convenient and comprehensive method for collecting wavefield information on complex curved CFRP parts for structural health monitoring, which would significantly improve quality control for CFRP components in various fields.

Tailored reinforcement architectures in discontinuous metal matrix composites (DMMCs) offer superior mechanical performance with broad scientific and financial interests. This study presents a domain-knowledge enhanced machine learning approach to efficiently explore the design space of Al-SiC DMMCs for optimization. A substantial dataset containing 140,000 instances, resembling characteristic reinforcement configurations and variants, is generated using a series of algorithms. Employing high-throughput finite element analysis, the elastic properties of each configuration are estimated. Statistical analysis reveals that a more homogeneous distributed reinforcement contributes to mechanical stability, whereas configurations with extreme performance tend to have inhomogeneous reinforcement distribution. A deep residual neural network trained on this dataset accurately learns the structure-property correlations. Coupled with a genetic algorithm, the framework identifies optimal configurations across different volume fractions for maximizing/minimizing properties including tensile modulus, shear modulus, and Poisson's ratio. Comparative analysis shows the incorporation of domain knowledge improves data quality, facilitating more effective design space exploration. This study contributes to advancing composite materials design, particularly for next-generation high-performance DMMCs.

In this study, we isolated and characterized HSP70 cDNA from pufferfish (Takifugu rubripes). The 3053 bp full-length TrHSP70sequence consisted of a 167 bp 5′-UTR (untranslated region), a 2535 bp open reading frame, and a 351 bp 3′-UTR. BLAST analysis revealed that the TrHSP70shared high similarity with HSP70 sequences in other species. In our study, we set 3 experimental groups as H1 group (20 °C), H2 group (24 °C), and H3 group (28 °C) for checking the expression level of TrHSP70in T. rubripes.Tissue-specific gene expression results showed that TrHSP70had higher expression in the intestines than other tissues of the T. rubripes byRT-qPCR. In the experimental group, we found that the expression of TrHSP70was upregulated in different tissues in the H3 group. The results show that TrHSP70is a constitutively expressed gene, which plays an important role in maintaining normal physiological function and coping with stress.Graphical AbstractThe complete cDNA sequence of the TrHSP70gene was cloned from T. rubripesby RACE-PCR. Gene expression of HSP70 was studied by RT-qPCR.

Cake layer formation is the dominant ultrafiltration membrane fouling mechanism after long-term operation. However, precisely analyzing the cake-layer structure still remains a challenge due to its thinness (micro/nano scale). Herein, based on the excellent depth-resolution and foulant-discrimination of time-of-flight secondary ion mass spectrometry, a three-dimensional analysis of the cake-layer structure caused by natural organic matter was achieved at lower nanoscale for the first time. When humic substances or polysaccharides coexisted with proteins separately, a homogeneous cake layer was formed due to their interactions. Consequently, membrane fouling resistances induced by proteins were reduced by humic substances or polysaccharides, leading to a high flux. However, when humic substances and polysaccharides coexisted, a sandwich-like cake layer was formed owing to the asynchronous deposition based on molecular dynamics simulations. As a result, membrane fouling resistances were superimposed, and the flux was low. Furthermore, it is interesting that cake-layer structures were relatively stable under common UF operating conditions (i.e., concentration and stirring). These findings better elucidate membrane fouling mechanisms of different natural-organic-matter mixtures. Moreover, it is demonstrated that membrane fouling seems lower with a more homogeneous cake layer, and humic substances or polysaccharides play a critical role. Therefore, regulating the cake-layer structure by feed pretreatment scientifically based on proven mechanisms should be an efficient membrane-fouling-control strategy.

Based on the theory of durability degradation of marine concrete, thorough investigation and analysis and field measurement of solid engineering components, the coupling test scheme of environment and load is formulated, and the pressure effect under different load levels and loading frequencies are studied. It is found that the chloride ion diffusion coefficient of concrete increases significantly with the increase of load level and the frequency of cyclic fatigue load increases. The results show that the chloride diffusion coefficient of structural concrete also increases significantly. The chloride diffusion coefficient of structural concrete with 2 and 5 Hz loading frequency conforms to the exponential relationship with the load level and the structural concrete with 10 Hz frequency conforms to the power relationship. Significantly reduce the service life of the concrete structure.

Alterations in lipid metabolites in coronary artery tissues are phenotypic changes in the progression of atherosclerosis (AS). A full picture of the spatiotemporal distribution of lipid metabolites in coronary AS is needed for a deeper understanding of its pathology and the identification of potential biomarkers of disease progression. In this work, the changes in species, quantity, and distribution of lipid metabolites at different stages of AS, which were standardized by the disease areas, were analyzed through the high spatial resolution- and high sensitivity-time-of-flight secondary ion mass spectrometry (ToF-SIMS) under delayed extraction mode. Based on high lateral resolution imaging, we further analyzed the ToF-SIMS data extracted from the subregions of AS lesion tissues at different disease progression stages by semiquantitative comparison, clustering analysis (t-stochastic neighbor embedding and HCA), and KEGG enrichment. Thus, a much-detailed description of lipids’ features in coronary AS was achieved. We constructed a ToF-SIMS mass spectrometry database of coronary AS lipids. 40 specific lipid metabolites with distinctive patterns between different pathological stages were obtained. Chemical imaging unveiled further details regarding the spatial distribution of lipids. Moreover, linoleic acid and arachidonic acid metabolic pathway were predicted to be critical in AS progression.

Central nervous system (CNS) lymphoma is an extranodal non-Hodgkin B-cell lymphoma characterized by malignant lymph tissue arising in the brain or spinal cord, associated with inflammation and blood-brain barrier (BBB) disruption. Although BBB disruption is known to occur in patients with CNS lymphoma, a direct link between these two has not been shown. Herein, abundant deposition of the blood coagulation protein fibrinogen around B-cell lymphoma was detected in CNS lymphoma patients and in the CNS parenchyma in an orthotopic mouse model. Functional enrichment analysis of unbiased cerebrospinal fluid proteomics of CNS B-cell lymphoma patients showed that coagulation protein networks were highly connected with tumor-associated biological signaling pathways. In vivotwo-photon imaging demonstrated that lymphoma growth was associated with BBB disruption, and in vitroexperiments identified a role for fibrinogen in promoting lymphoma cell adhesion. Overall, these results identify perivascular lymphoma clustering at sites of fibrinogen deposition, and suggest that fibrinogen may be a target for pharmacologic intervention in metastatic B-cell lymphoma associated with BBB disruption.

Cutaneous T-cell lymphoma is a form of non-Hodgkin lymphoma that manifests initially in the skin and disseminates systemically as the disease progresses. Mycosis fungoides and Sézary syndrome are the most common subtypes of cutaneous T-cell lymphoma. Advanced mycosis fungoides and Sézary syndrome are life threatening with few treatment options. We searched for new agents by high-throughput screening of selected targeted compounds and identified high-value targets, including phosphatidylinositol 3-kinase (PI3K) and cyclin-dependent kinases. To validate these hits from the screen, we developed patient-derived xenograft mouse models that recapitulated the cardinal features of mycosis fungoides and Sézary syndrome and maintained histologic and molecular characteristics of their clinical counterparts. Importantly, we established a blood-based biomarker assay using tumor cell-free DNA to measure systemic tumor burden longitudinally in living mice during drug therapy. A PI3K inhibitor, BKM120, was tested in our patient-derived xenograft model leading to disease attenuation and prolonged survival. Isoform-specific small interfering RNA knockdowns and isoform-selective PI3K inhibitors identified PI3K-δ as required for tumor proliferation. Additional studies showed a synergistic combination of PI3K-α/δ inhibitors with histone deacetylase inhibitors. The strong preclinical efficacy of this potent combination against multiple patient-derived xenograft models makes it an excellent candidate for further clinical development.

Insect resistance to insecticides is an increasingly serious problem, and the resistant mechanisms are complicated. The resistance research based on the chemosensory pathway is one of the hot problems at present, but the specific binding mechanism of chemosensory genes and insecticides remains elusive. The binding mechanism of AlepGOBP2 (belong to insect chemosensory gene) with two insecticides was investigated by computational and experimental approaches. Our calculation results indicated that four key residues (Phe12, Ile52, Ile94, and Phe118) could steadily interact with these two insecticides and be assigned as hotspot sites responsible for their binding affinities. The significant alkyl−π and hydrophobic interactions involved by these four hotspot residues were found to be the driving forces for their binding affinities, especially for two residues (Phe12 and Ile94) that significantly contribute to the binding of chlorpyrifos, which were also validated by our binding assay results. Furthermore, we also found that the AlepGOBP2–chlorpyrifos/phoxim complexes can be more efficiently converged in the residue-specific force field-(RSFF2C) and its higher accuracy and repeatability in protein dynamics simulation, per-residue free energy decomposition, and computational alanine scanning calculations have also been achieved in this paper. These findings provided useful insights for efficient and reliable calculation of the binding mechanism of relevant AlepGOBPs with other insecticides, facilitating to develop new and efficient insecticides targeting the key sites of AlepGOBP2.

Hydroxyl radical (•OH) can hydroxylate or dehydrogenate organics without forming extra products and is thereby expediently applied in extensive domains. Although it can be efficiently produced through single-electron transfer from transition-metal-containing activators to hydrogen peroxide (H2O2), narrow applicable pH range, strict activator/H2O2ratio requirement, and byproducts that are formed in the mixture with the background matrix necessitate the need for additional energy-intensive up/downstream treatments. Here, we show a green Fenton process in an electrochemical cell, where the electro-generated atomic H* on a Pd/graphite cathode enables the efficient conversion of H2O2into •OH and subsequent degradation of organic pollutants (80% efficiency). Operando liquid time-of-fight secondary ion mass spectrometry verified that H2O2activation takes place through a transition state of the Pd–H*–H2O2adduct with a low reaction energy barrier of 0.92 eV, whereby the lone electron in atomic H* can readily cleave the peroxide bridge, with •OH and H2O as products (ΔGr= −1.344 eV). Using H+or H2O as the resource, we demonstrate that the well-directed output of H* determines the pH-independent production of •OH for stable conversion of organic contaminants in wider pH ranges (3–12). The research pioneers a novel path for eliminating the restrictions that are historically challenging in the traditional Fenton process.

Real-time in situ imaging of organelles is increasingly important in modern biomedical analysis and diseases diagnosis. To realize this goal, organelle-targeting nanoparticles as one of the most commonly used technologies in subcellular sensing and imaging has attracted a lot of interest. The biocompatibility, specificity, and binding efficiency are especially critical for efficient organelle-targeting bioimaging. Gold nanoparticles (AuNPs) fabricated with bifunctional peptides constructed with both Au-binding affinity and nucleus-targeting ability were designed and examined for efficient nucleus-targeting bioimaging. Such a design is expected to achieve an oriented assembling of peptides by the medium of the Au-binding peptides specifically assembled on the surface of AuNPs, with the nucleus-targeting end open for accessibility. The bifunctional peptides showed strong binding affinity toward AuNPs and led to a binding capability ∼1.5 times higher than that of the bare nucleus-targeting peptides, ensuring good surface coverage of the nanoparticles for enhanced nucleus-targeting ability. Such fabricated AuNPs demonstrated over 90% cell viability after incubation for 24 h with HepG2 cells, which were highly biocompatible. Precise and efficient bioimaging of the nucleus was achieved for HepG2 cells by using the fabricated AuNPs as observed with a confocal laser scanning microscope, a dark-field/fluorescence microscope, and a transmission electron microscope. The high surface coverage and oriented binding pattern appeared to be a promising strategy for construction of organelle-targeting agencies.

Ferromagnetic microwires are feasible building blocks for the construction of electromagnetic composites with tunable left-handed response towards external stimulus. In this work, a typical left-handed signature of the transmission window was revealed in Co-rich microwires ranging from 9.3 to 10.4 GHz, which arised from the ferromagnetic resonance under a constant DC magnetic field of 8 Oe. Such window was modulated by applying a DC current which tended to compensate the magnetic anisotropy of the microwires. Currents of up to 100 mA were applied to the wire resulting in a maximum transmission bandwidth at 60 mA. Such effect was corroborated by the competition between magnetic domains restoration and Joule effect in the presence of DC current. Embedding the microwire into fibre-reinforced 913 E-glass prepregs realised a metacomposite with a corresponding window in 8.5–10.2 GHz benefited by the restoration of circumferential domain structures during the composite curing process. Further, it was observed a dependence of electromagnetic behaviour on the angle between the microwire and the reinforcing glass fibers of the matrix. This investigation provides essential guidance for designing and tuning frequency-selective wave transmitting structures for potential sensing and cloaking applications.

A goal of HIV vaccine development is to elicit antibodies with neutralizing breadth. Broadly neutralizing antibodies (bNAbs) to HIV often have unusual sequences with long heavy-chain complementarity-determining region loops, high somatic mutation rates and polyreactivity. A subset of HIV-infected individuals develops such antibodies, but it is unclear whether this reflects systematic differences in their antibody repertoires or is a consequence of rare stochastic events involving individual clones. We sequenced antibody heavy-chain repertoires in a large cohort of HIV-infected individuals with bNAb responses or no neutralization breadth and uninfected controls, identifying consistent features of bNAb repertoires, encompassing thousands of B cell clones per individual, with correlated T cell phenotypes. These repertoire features were not observed during chronic cytomegalovirus infection in an independent cohort. Our data indicate that the development of numerous B cell lineages with antibody features associated with autoreactivity may be a key aspect in the development of HIV neutralizing antibody breadth.

Electrode-electrolyte interface, where the electrochemical reaction happened, played an important role in the electrochemical mechanism investigation. The traditional electrochemical techniques paid more attention on the electron transfer process on the electrode but provided limited information of the electrochemical evolution at molecular level. Realizing the in-situmonitoring of the intermediates at the electrode-electrolyte interface during the redox reaction was essential for the study of electrochemical mechanism. In this work, electrochemical reaction of coenzyme Q0(CoQ0) on gold electrode was investigated by time-of-flight secondary ion mass spectrometry (ToF-SIMS). A high vacuum compatible microfluidic electrochemical cell was fabricated for the electrochemical study of CoQ0, which realized the in-situdetection of the electrode-electrolyte interface in high vacuum environment (10−5Pa). The variations of CoQ0, CoQ0H2and related intermediate at different voltages agreed well with the electrochemical reaction of CoQ0in aqueous solution. The direct monitoring of electrode-electrolyte interface by pre-confined ToF-SIMS could reveal the electrochemical evolution on the electrode at molecular level, which was of great potential for the detailed understanding of the electrochemical reaction mechanism.

Epsilon-near-zero (ENZ) materials have drawn considerable attention due to their enticing properties, such as large wavelength, quasi-static and tunneling. Most research on ENZ behaviors is conducted in the visible, microwave, and infrared bands, but there is seldom research in radio frequency (MHz∼GHz). In this work, polyimide (PI) embedded cobalt nanoparticles (NPs) were fabricated to realize ENZ behavior in MHz-bands. The percolation networks were constructed and the dielectric resonance frequency was observed for PI-35 wt% Co composite at 580 MHz. As Co content increased, the ENZ response was obtained due to dielectric resonance and plasma oscillation. Also, conduction mechanism of PI/Co composites undergo a change from hopping state to metal-like state. The reactance value changed from negative to positive with increasing frequency, corresponding to the transition of permittivity from positive to negative value.

A sodium-ethylenediamine graphite intercalation compound (Na(ethylenediamine)C15: “GIC”) made from graphite flakes was used to study the microwave absorption performance of a GIC for the first time. Compared with the pristine graphite flakes, the neighboring layers in this GIC are pillared by Na(ethylenediamine)+and possess a larger layer distance and improved electrical conductivity. Owing to the electrical conductivity of this GIC, only half of the loading content, compared to graphite flakes, is needed to achieve an outstanding absorption of −75.6 dB at 9.25 GHz (10.0 wt % GIC in paraffin in a 4.0 mm thick sample), but for graphite, 20.0 wt % is required for an absorption of −37.6 dB.

Triterpene esters comprise a class of secondary metabolites that are synthesized by decorating triterpene skeletons with a series of oxidation, glycosylation, and acylation modifications. Many triterpene esters with important bioactivities have been isolated and identified, including those with applications in the pesticide, pharmaceutical, and cosmetic industries. They also play essential roles in plant defense against pests, diseases, physical damage (as part of the cuticle), and regulation of root microorganisms. However, there has been no recent summary of the biosynthetic pathways and biological functions of plant triterpene esters. Here, we classify triterpene esters into five categories based on their skeletons and find that C-3 oxidation may have a significant effect on triterpenoid acylation. Fatty acid and aromatic moieties are common ligands present in triterpene esters. We further analyze triterpene ester synthesis-related acyltransferases (TEsACTs) in the triterpene biosynthetic pathway. Using an evolutionary classification of BAHD acyltransferases (BAHD-ATs) and serine carboxypeptidase-like acyltransferases (SCPL-ATs) in Arabidopsis thalianaand Oryza sativa, we classify 18 TEsACTs with identified functions from 11 species. All the triterpene-skeleton-related TEsACTs belong to BAHD-AT clades IIIa and I, and the only identified TEsACT from the SCPL-AT family belongs to the CP-I subfamily. This comprehensive review of the biosynthetic pathways and bioactivities of triterpene esters provides a foundation for further study of their bioactivities and applications in industry, agricultural production, and human health.

Hygrothermal effects on the translaminar fracture toughness of the carbon/epoxy composite with two different stacking sequences at 120 °C were studied and the failure mechanisms are explained. The eccentrically loaded single-edge-notch tension (ESET) experiments showed that two different quasi-isotropic stacking sequences ([90/45/0/-45]4sand [45/90/-45/0]4s) have negligible difference in translaminar fracture toughness. It was found that the fracture toughness of conditioned specimens at hot temperature wet (HTW) condition increased by 75 % compared to that at room temperature dry (RTD). The average saturated damage height at HTW increased by 48 % compared with that at RTD according to ex situX-ray computed tomography (CT) observation. The larger saturated damage height observed in the 0° plies of the HTW specimens results in the higher translaminar fracture toughness.

Exosomes, also termed extracellular vesicles (EVs), are an important component of the tumor microenvironment (TME) and exert versatile effects on the molecular communications in the TME of hepatocellular carcinoma (HCC). Exosome-mediated intercellular communication is closely associated with the tumorigenesis and development of HCC. Exosomes can be extracted through ultracentrifugation and size exclusion, followed by molecular analysis through sequencing. Increasing studies have confirmed the important roles of exosome-derived ncRNAs in HCC, including tumorigenesis, progression, immune escape, and treatment resistance. Due to the protective membrane structure of exosomes, the ncRNAs carried by exosomes can evade degradation by enzymes in body fluids and maintain good expression stability. Thus, exosome-derived ncRNAs are highly suitable as biomarkers for the diagnosis and prognostic prediction of HCC, such as exosomal miR-21-5p, miR-221-3p and lncRNA-ATB. In addition, substantial studies revealed that the up-or down-regulation of exosome-derived ncRNAs had an important impact on HCC progression and response to treatment. Exosomal biomarkers, such as miR-23a, lncRNA DLX6-AS1, miR-21-5p, lncRNA TUC339, lncRNA HMMR-AS1 and hsa_circ_0004658, can reshape immune microenvironment by regulating M2-type macrophage polarization and then promote HCC development. Therefore, by controlling exosome biogenesis and modulating exosomal ncRNA levels, HCC may be inhibited or eliminated. In this current review, we summarized the recent findings on the role of exosomes in HCC progression and analyzed the relationship between exosome-derived ncRNAs and HCC diagnosis and treatment.

Graphene nanoplates are hoped-for solid lubricants to reduce friction and energy dissipation in micro and nanoscale devices benefiting from their interface slips to reach an expected superlubricity. On the contrary, we propose here by introducing engineered wrinkles of graphene nanoplates to exploit and optimize the interfacial energy dissipation mechanisms between the nanoplates in graphene-based composites for enhanced vibration damping performance. Polyurethane (PU) beams with designed sandwich structures have been successfully fabricated to activate the interlaminar slips of wrinkled graphene–graphene, which significantly contribute to the dissipation of vibration energy. These engineered composite materials with extremely low graphene content (∼0.08 wt %) yield a significant increase in quasi-static and dynamic damping compared to the baseline PU beams (by 71% and 94%, respectively). Friction force images of wrinkled graphene oxide (GO) nanoplates detected via an atomic force microscope (AFM) indicate that wrinkles with large coefficients of friction (COFs) indeed play a dominant role in delaying slip occurrences. Reduction of GO further enhances the COFs of the interacting wrinkles by 7.8%, owing to the increased effective contact area and adhesive force. This work provides a new insight into how to design graphene-based composites with optimized damping properties from the microstructure perspective.

A pore confined liquid–vacuum interface was created in liquid secondary ion mass spectrometry analysis in order to study the charge transfer in electrochemical reactions. The interfacial processes such as the critical diameter, influence of aperture properties on the morphology of the liquid–vacuum interface, pressure field, concentration field, and electric field were revealed by finite element simulation. The correlation between numerical study of the chemical changes at the electrode–electrolyte interface and experimental results during the dynamic potential scan was built successfully. Better understanding of these interfacial processes could promote further applications of liquid secondary ion mass spectrometry in energy storage and electrochemical catalysis.

This short work is to communicate the influence of hygrothermal aging on the mechanical performances of fiber-metal-laminates (FMLs) with various ingenious metal surface treatments, namely, nano-scale electrochemical sculpture (NES), the conventional surface polishing (PS) and NaTESi anodizing (NaTESi). The specimens with different edge conditions were prepared using milling or wire-electrode cutting and subjected to hygrothermal aging treatment in a climatic test chamber at 60 °C and 96% relative humidity. Interlaminar shear and flexural testing results showed that the wire-electrode cutting method produced edges with exposed fibers and spalling resin after hygrothermal aging, while the milling cutting method resulted in edges with a micro-porous and defective surface, but without exposing fibers or spalling resin. The degree of reduction in interlaminar shear strength (ILSS) and flexural strength varied depending on the surface treatments and machining methods used. After undergoing hygrothermal aging, the wire-electrode cutting method causes a greater decrease in ILSS compared to the milling cutting method. This reduction is particularly pronounced in specimens with weaker interfacial bonding strength, in which the ILSS of wire-electrode cut FMLs-PS decreased by 32.5%, while that of FMLs-NES decreased by 17.8%.

A library consisting of 429 food-source compounds was used to screen the natural products with anticancer properties in esophageal squamous cell carcinoma (ESCC). We demonstrated for the first time that synephrine, an active compound isolated from leaves of citrus trees, markedly suppressed cell proliferation (inhibition rate with 20 μM synephrine at day 5:71.1 ± 5.8% and 75.7 ± 6.2% for KYSE30 and KYSE270, respectively) and colony formation (inhibition rate with 10 μM synephrine: 86.5 ± 5.9% and 82.3 ± 4.5% for KYSE30 and KYSE270, respectively), as well as migration (inhibition rate with 10 μM synephrine: 76.9 ± 4.4% and 62.2 ± 5.8% for KYSE30 and KYSE270, respectively) and invasion abilities (inhibition rate with 10 μM synephrine: 73.3 ± 7.5% and 75.3 ± 3.4% for KYSE30 and KYSE270, respectively) of ESCC cells in a dose-dependent manner, without significant toxic effect on normal esophageal epithelial cells. Mechanistically, quantitative proteomics and bioinformatics analyses were performed to explore the synephrine-regulated proteins. Western blot and qRT-PCR data indicated that synephrine may downregulate Galectin-3 to inactivate AKT and ERK pathways. In addition, we found that the sensitivity of ESCC to fluorouracil (5-FU) could be enhanced by synephrine. Furthermore, in vivo experiments showed that synephrine had significant antitumor effect on ESCC tumor xenografts in nude mice (inhibition rate with 20 mg/kg synephrine is 61.3 ± 20.5%) without observed side effects on the animals. Taken together, synephrine, a food-source natural product, may be a potential therapeutic strategy in ESCC.

T follicular helper (Tfh) cells are required to develop germinal center (GC) responses and drive immunoglobulin class switch, affinity maturation, and long-term B cell memory. In this study, we characterize a recently developed vaccine platform, nucleoside-modified, purified mRNA encapsulated in lipid nanoparticles (mRNA-LNPs), that induces high levels of Tfh and GC B cells. Intradermal vaccination with nucleoside-modified mRNA-LNPs encoding various viral surface antigens elicited polyfunctional, antigen-specific, CD4+ T cell responses and potent neutralizing antibody responses in mice and nonhuman primates. Importantly, the strong antigen-specific Tfh cell response and high numbers of GC B cells and plasma cells were associated with long-lived and high-affinity neutralizing antibodies and durable protection. Comparative studies demonstrated that nucleoside-modified mRNA-LNP vaccines outperformed adjuvanted protein and inactivated virus vaccines and pathogen infection. The incorporation of noninflammatory, modified nucleosides in the mRNA is required for the production of large amounts of antigen and for robust immune responses.

Development of a highly effective vaccine or antibodies for the prevention and ultimately elimination of malaria is urgently needed. Here we report the isolation of a number of human monoclonal antibodies directed against the Plasmodium falciparum (Pf) circumsporozoite protein (PfCSP) from several subjects immunized with an attenuated Pf whole-sporozoite (SPZ) vaccine (Sanaria PfSPZ Vaccine). Passive transfer of one of these antibodies, monoclonal antibody CIS43, conferred high-level, sterile protection in two different mouse models of malaria infection. The affinity and stoichiometry of CIS43 binding to PfCSP indicate that there are two sequential multivalent binding events encompassing the repeat domain. The first binding event is to a unique 'junctional' epitope positioned between the N terminus and the central repeat domain of PfCSP. Moreover, CIS43 prevented proteolytic cleavage of PfCSP on PfSPZ. Analysis of crystal structures of the CIS43 antigen-binding fragment in complex with the junctional epitope determined the molecular interactions of binding, revealed the epitope's conformational flexibility and defined Asn-Pro-Asn (NPN) as the structural repeat motif. The demonstration that CIS43 is highly effective for passive prevention of malaria has potential application for use in travelers, military personnel and elimination campaigns and identifies a new and conserved site of vulnerability on PfCSP for next-generation rational vaccine design.

The size distribution and molecular structure of water clusters play a critical role in the chemical, biological and atmospheric process. The common experimental study of water clusters in aqueous solution is challenged due to the influence of local Hbonding environments on vibration spectroscopies or vacuum requirements for most mass spectrometry technologies. Here, the time-of-flight secondary ion mass spectrometry (ToF-SIMS) combining with a microfluidic chip has been applied to achieve the in-situdiscrimination of the size distribution for water clusters in liquid water at room temperature. The results demonstrated that the presented method is highly system stable, reproducible and accurate. The comparison of heavy water with pure water was made to further demonstrate the accuracy of this technique. These results showed that (H2O)3H+and (D2O)4D+are the most dominant clusters in pure and heavy water, respectively. This one water molecule difference in the dominant cluster size may due to the nuclear quantum effects on water’s hydrogen bonded network. It is the first time to experimentally show the size distribution of water clusters over a wide range (n=1–30) for pure (H2O) and heavy (D2O) water from molecular level. This technique provides an achievable method for liquid water, which offers a bridge to close the gap between theoretical and experimental study of water cluster in aqueous solution.

The chemical components analysis of single cell is important for the understanding of physiological processes such as cell growth, signal transduction and apoptosis. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a sensitive surface analysis technique with high spatial resolution, which has been used for single cell and micro-area analysis. However, relatively low ionization yield of biomolecules limited its wide applications in single cell analysis. Herein, we used metal substrate and matrix material to enhance the ionization yield of lipids. The signal intensity of phosphatidylcholine (PC 40:0) casted on the matrix/gold-coated silicon substrate was 65 times higher than that on the silicon wafer. The signal enhancement of phosphatidylcholine (PC 34:1) on single cell surface cultured on matrix/gold-coated silicon substrate was observed as well. Owing to the influence of irregular topography and complex chemical environment of cell, the increase of lipids signal was smaller. Delayed extraction mode of ToF-SIMS overcame the effects of cell topography, leading to further enhancement of the signal intensity of lipids. Meanwhile, simultaneous high spatial resolution of chemical imaging and high mass resolution of the mass spectra of single cells were obtained. Our strategies provided new insights into the study of cell metabolism and cell-environment interactions.

In this study, a safety enhanced SalmonellaPullorum (S. Pullorum) ghost was constructed using an antimicrobial peptide gene, and evaluated for its potential as a Pullorum disease (PD) vaccine candidate. The antimicrobial peptide SMAP29 was co-expressed with lysis gene E to generate S. Pullorum ghosts. No viable bacteria were detectable either in the fermentation culture after induction of gene E- and SMAP29-mediated lysis for 24 h or in the lyophilized ghost products. Specific-pathogen-free (SPF) chicks were intraperitoneally immunized with ghosts at day 7 of age and no mortality, clinical symptoms or signs of PD such as anorexia, depression and diarrhea were observed. On challenge with a virulent S. Pullorum strain at 4 wk post-immunization, a comparatively higher level of protection was observed in the S. Pullorum ghost immunized chickens with a minimum of pathological lesions and bacterial loads compared to the birds in inactivated vaccine groups. In addition, immunization with the S. Pullorum ghosts induced a potent systemic IgG response and was associated with significantly increased levels of cytokine IFN-γ and IL-4 and relative percentages of CD4+and CD8+T lymphocytes. Our results indicate that SMAP29 can be employed as a new secondary lethal protein to enhance the safety of bacterial ghosts, and to prepare a non-living bacterial vaccine candidate that can prevent PD in chickens.