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Summary 1. Kindlin-3 but not talin-1 contributes to integrin inside-out signaling induced β2 integrin clustering. 2. The Pleckstrin homology domain of kindlin-3 was critical for its mediated β2 integrin clustering.

Abstract To evaluate the room-temperature hydrogen embrittlement susceptibility (HES) of high-pressure hydrogen storage vessels, a modified slow-strain-rate tensile (MSSRT) testing method was proposed for effectively aligning with their actual operating conditions. The effectiveness of the MSSRT testing method in evaluating the room-temperature HES of steels under high pressure was validated by comparing the results obtained using the conventional slow strain rate tensile (SSRT) and proposed MSSRT testing methods for 30CrMo steel, which is widely used for manufacturing high-pressure hydrogen storage vessels. The tensile properties and fracture morphologies of 23Cr2Ni4MoV steel were then examined using the MSSRT testing method under 35 MPa hydrogen and nitrogen at room temperature. Results indicate that 35 MPa hydrogen exerted a marginal effect on the tensile properties of 23Cr2Ni4MoV steel at room temperature when considering the MSSRT testing method; moreover, the test specimen basically exhibited macroscopic ductile fracture. Furthermore, obvious surface cracking was observed on the fractured specimen tested under hydrogen, whereas surface cracking was not observed on the fractured specimen tested under nitrogen. Hence, the relative reduction of area and surface cracking are necessary criteria for evaluating the room-temperature HES of steels using the MSSRT testing method. Overall, 23Cr2Ni4MoV steel might be unsuitable for manufacturing high-pressure hydrogen storage vessels.

Abstract Arterial thrombosis is a leading cause of death and disability worldwide with no effective bioassay for clinical prediction. As a symbolic feature of arterial thrombosis, severe stenosis in the blood vessel creates a high-shear, high-gradient flow environment that facilitates platelet aggregation towards vessel occlusion. Here, we present a thrombus profiling assay that monitors the multi-dimensional attributes of thrombi forming in such biomechanical conditions. Using this assay, we demonstrate that different receptor–ligand interactions contribute distinctively to the composition and activation status of the thrombus. Our investigation into hypertensive and older individuals reveals intensified biomechanical thrombogenesis and multi-dimensional thrombus profile abnormalities, endorsing the diagnostic potential of the assay. Furthermore, we identify the hyperactivity of GPIbα-integrin αIIbβ3 mechanosensing axis as a molecular mechanism that contributes to hypertension-associated arterial thrombosis. By studying drug-disease interactions and inter-individual variability, our work reveals a need for personalized anti-thrombotic drug selection that accommodates each patient’s pathological profile.

To ensure optimal penetration in direct current (DC) argon tungsten arc welding, a method was proposed to recognize the molten pool penetration during molten pool oscillation under alternating cusp-shaped magnetic field (ACSMF). An arc discharge model is established to analyze the oscillation mechanism of the molten pool under ACSMF. The periodic variation of arc shape induces the periodic oscillation to establish a mapping relationship with the fluctuation of arc voltage waveform. At the excitation current of 3 A and frequency of 10 Hz, the arc voltage waveform undergoes abrupt changes at 1.26 V and 1.37 V, corresponding to the critical and the full penetration states, respectively. The errors in identifying moving penetration are 0.8 % and 0.7 %, with a full penetration weld, demonstrating a penetration rate of 100 %, indicating effective penetration recognition. Arc sensing technology for tungsten inert gas (TIG) welding under ACSMF is poised for gradual application in single-side welding and double-side forming of medium and thick plates. It is anticipated to evolve towards real-time identification technology for molten pool penetration monitoring.

Abstract The Phosphatidylinositol-3-kinase (PI3K) family is well-known to comprise three classes of intracellular enzymes. Class I PI3Ks primarily function in signaling by responding to cell surface receptor stimulation, while class II and III are more involved in membrane transport. Under normal physiological conditions, the PI3K signaling network orchestrates cell growth, division, migration and survival. Aberrant activation of the PI3K signaling pathway disrupts cellular activity and metabolism, often marking the onset of cancer. Currently, the Food and Drug Administration (FDA) has approved the clinical use of five class I PI3K inhibitors. These small-molecule inhibitors, which exhibit varying selectivity for different class I PI3K family members, are primarily used in the treatment of breast cancer and hematologic malignancies. Therefore, the development of novel class I PI3K inhibitors has been a prominent research focus in the field of oncology, aiming to enhance potential therapeutic selectivity and effectiveness. In this review, we summarize the specific structures of PI3Ks and their functional roles in cancer progression. Additionally, we critically evaluate small molecule inhibitors that target class I PI3K, with a particular focus on their clinical applications in cancer treatment. Moreover, we aim to analyze therapeutic approaches for different types of cancers marked by aberrant PI3K activation and to identify potential molecular targets amenable to intervention with small-molecule inhibitors. Ultimately, we propose future directions for the development of therapeutic strategies that optimize cancer treatment outcomes by modulating the PI3K family.

Abstract Sirtuin 3 (SIRT3) is well known as a conserved nicotinamide adenine dinucleotide+ (NAD+)-dependent deacetylase located in the mitochondria that may regulate oxidative stress, catabolism and ATP production. Accumulating evidence has recently revealed that SIRT3 plays its critical roles in cardiac fibrosis, myocardial fibrosis and even heart failure (HF), through its deacetylation modifications. Accordingly, discovery of SIRT3 activators and elucidating their underlying mechanisms of HF should be urgently needed. Herein, we identified a new small-molecule activator of SIRT3 (named 2-APQC) by the structure-based drug designing strategy. 2-APQC was shown to alleviate isoproterenol (ISO)-induced cardiac hypertrophy and myocardial fibrosis in vitro and in vivo rat models. Importantly, in SIRT3 knockout mice, 2-APQC could not relieve HF, suggesting that 2-APQC is dependent on SIRT3 for its protective role. Mechanically, 2-APQC was found to inhibit the mammalian target of rapamycin (mTOR)-p70 ribosomal protein S6 kinase (p70S6K), c-jun N-terminal kinase (JNK) and transforming growth factor-β (TGF-β)/ small mother against decapentaplegic 3 (Smad3) pathways to improve ISO-induced cardiac hypertrophy and myocardial fibrosis. Based upon RNA-seq analyses, we demonstrated that SIRT3-pyrroline-5-carboxylate reductase 1 (PYCR1) axis was closely assoiated with HF. By activating PYCR1, 2-APQC was shown to enhance mitochondrial proline metabolism, inhibited reactive oxygen species (ROS)-p38 mitogen activated protein kinase (p38MAPK) pathway and thereby protecting against ISO-induced mitochondrialoxidative damage. Moreover, activation of SIRT3 by 2-APQC could facilitate AMP-activated protein kinase (AMPK)-Parkin axis to inhibit ISO-induced necrosis. Together, our results demonstrate that 2-APQC is a targeted SIRT3 activator that alleviates myocardial hypertrophy and fibrosis by regulating mitochondrial homeostasis, which may provide a new clue on exploiting a promising drug candidate for the future HF therapeutics.

Summary: Background: The ubiquitin regulatory X (UBX) domain-containing proteins (UBXNs) are putative adaptors for ubiquitin ligases and valosin-containing protein; however, their in vivo physiological functions remain poorly characterised. We recently showed that UBXN3B is essential for activating innate immunity to DNA viruses and controlling DNA/RNA virus infection. Herein, we investigate its role in adaptive immunity. Methods: We evaluated the antibody responses to multiple viruses and pathogenesis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza in tamoxifen-inducible global and constitutive B cell-specific Ubxn3b knockout mice; quantified various immune populations, B lineage progenitors/precursors, B cell receptor (BCR) signalling and apoptosis by flow cytometry, immunoblotting and immunofluorescence microscopy. We also performed bone marrow transfer, single-cell and bulk RNA sequencing. Findings: Both global and B cell-specific Ubxn3b knockout mice present a marked reduction in small precursor B-II (>60%), immature (>70%) and mature B (>95%) cell numbers. Transfer of wildtype bone marrow to irradiated global Ubxn3b knockouts restores normal B lymphopoiesis, while reverse transplantation does not. The mature B population shrinks rapidly with apoptosis and higher pro and activated caspase-3 protein levels were observed following induction of Ubxn3b knockout. Mechanistically, Ubxn3b deficiency leads to impaired pre-BCR signalling and cell cycle arrest. Ubxn3b knockout mice are highly vulnerable to respiratory viruses, with increased viral loads and prolonged immunopathology in the lung, and reduced production of virus-specific IgM/IgG. Interpretation: UBXN3B is essential for B lymphopoiesis by maintaining constitutive pre-BCR signalling and cell survival in a cell-intrinsic manner. Funding: United States National Institutes of Health grants, R01AI132526 and R21AI155820.

To determine the structure and sequence variation of the chloroplast genome of genus Aucuba and to reveal their interspecies relationship, the chloroplast genomes of six Aucuba species including A. chinensis and A. japonica var. variegata as well as Garrya buxifolia were sequenced using next-generation sequencing technology. The chloroplast genomes were assembled and annotated using bioinformatics software. The genome characteristics, sequence comparison and phylogeny were analyzed based on the chloroplast genomes. The results were as follows: (1) Six Aucuba species chloroplast genomes exhibited typical quadripartite circular structure (i.e., LSC, SSC, IRB and IRA) and were between 157 891-158 325 bp in length. A total of 114 genes were annotated (except the duplicate genes on IRs), including 80 protein-coding genes (CDS), 30 tRNA genes and 4 rRNA genes. (2) A total of 29 high-frequency codons prefer ending in A/U, and a total of 100 optimal codons of these six sequences were filtered out, including 12 were shared by six Aucuba species. (3) A total of 270 interspersed repeats, 133 tandem repeats and 412 SSR loci were detected in the six Aucuba chloroplast genomes. (4) Comparative analysis result of six Aucuba species revealed a highly conserved chloroplast genome sequence. (5) Ten highly divergent regions were identified based on nucleotide diversity analysis. (6) The phylogenetic trees were constructed using maximum likelihood (ML) and Bayesian (BI) showing a similar topology. The results supported that genus Aucuba was monophyletic with high support rate, as a sister to genus Garrya. The complete chloroplast genomes of these five Aucuba species and one Garrya species is firstly sequenced in the study. The results reveal the interspecies relationships and the taxonomic status of genus Aucuba, and provide references for the classification, identification and phylogeny of the genus Aucuba.

In order to address the ‘capacity crisis’ caused by the narrow bandwidth of the current C band and the demand for wide-spectrum sensing sources and tunable fiber lasers, a broadband luminescence covering the C + L bands using Er3+/Yb3+ co-doped fluorotellurite glass fiber is investigated in this paper. The optimal doping concentrations in the glass host were determined based on the intensity, lifetime, and full width at half maximum (FWHM) of the fluorescence centered at 1.5 µm, which were found to be 1.5 mol% Er2O3 and 3 mol% Yb2O3. We also systematically investigated this in terms of optical absorption spectra, absorption and emission cross-sections, gain coefficients, Judd–Ofelt parameters, and up-conversion fluorescence. The energy transfer (ET) mechanism between the high concentrations of Er3+ and Yb3+ was summarized. In addition, a step-indexed fiber was prepared based on these fluorotellurite glasses, and a wide bandwidth of ~112.5 nm (covering the C + L bands from 1505.1 to 1617.6 nm) at 3 dB for the amplified spontaneous emission (ASE) spectra has been observed at a fiber length of 0.57 m, which is the widest bandwidth among all the reports based on tellurite glass. Therefore, this kind of Er3+/Yb3+ co-doped fluorotellurite glass fiber has great potential for developing broadband C + L band amplifiers, ultra-wide fiber sources for sensing, and tunable fiber lasers.

Peritonitis is a common and life-threatening inflammatory disease. Myeloid cells are elevated in the peripheral blood and contribute to peritonitis, but their circulating dynamics are not clear. In vivo flow cytometry (IVFC) is a noninvasive technique for monitoring the dynamics of circulating cells in live animals. It has been extensively used to detect circulating tumor cells, but rarely for monitoring immune cells. Here, we describe a method adapting an intravital microscope for IVFC so that we can monitor LysM-EGFP-labeled circulating myeloid cells in a tumor necrosis factor (TNF) α-induced peritonitis mouse model. Using this IVFC method, we quantified the blood flow velocity and cell concentration in circulation. We observed a significant increase in LysM-EGFP+ cells in circulation after TNFα intraperitoneal (i.p.) injection, which reached a plateau in ~20 min. Conventional cytometry analysis showed that most LysM-EGFP+ cells were neutrophils. Increasing blood neutrophils were accompanied by neutrophil recruitment to the peritoneal cavity and neutrophil emigration from the bone marrow. We then monitored neutrophil CD64 expression in vivo and found a significant increase in TNFα-induced peritonitis. We also found that CD18 blockade doubled the circulating neutrophil number in TNFα-induced peritonitis, suggesting that CD18 is critical for neutrophil recruitment in peritonitis. Overall, we demonstrate that IVFC techniques are useful for studying the circulating dynamics of immune cells during inflammatory diseases.

In the petrochemical industry, centrifugal cast Fe–Cr–Ni heat-resistant alloy tubes are critical components that are exposed to severe conditions for long-time service. The life of the alloy tubes is primarily limited by creep damages, which can evolve into catastrophic failures. In order to improve the creep properties in a low-cost approach, the influence of N + C content on the microstructure evolution and creep behavior of the 25Cr35NiNb alloys has been studied in this work. Creep tests were carried out at 1173 K (900 °C) and 1323 K (1050 °C) with different stress levels. Microstructure investigation on precipitates revealed that the C/N ratio significantly affects the content and morphology of Cr-rich carbides and the phase transition of Nb-rich carbonitrides after thermal exposure. By adopting the threshold stress analysis, the true creep activation energy and stress exponent of all the alloys were found to be ∼270 kJ mol−1 and ∼5, respectively, which indicated that dislocation climb is the rate-controlling mechanism of creep. Furthermore, the tailored novel N-bearing alloy possessed superior comprehensive properties of creep resistance and damage tolerance compared to the 25Cr35NiNb grade alloys. Our current findings not only realize improved creep properties by the combined modification of N addition and C/N ratio, but also provide new insights into understanding the microalloying mechanisms of high-temperature materials in general.

Summary: Canonical interleukin-2 (IL-2) signaling via the high-affinity CD25-containing IL-2 receptor-Janus kinase (JAK)1,3-signal transducer and activator of transcription 5 (STAT5) pathway is essential for development and maintenance of CD4+CD25HiFoxp3+ regulatory T cells (Tregs) that support immune homeostasis. Here, we report that IL-2 signaling via an alternative CD25-chemokine receptor pathway promotes the suppressive function of Tregs. Using an antibody against CD25 that biases IL-2 signaling toward this alternative pathway, we establish that this pathway increases the suppressive activity of Tregs and ameliorates murine experimental autoimmune encephalomyelitis (EAE). Furthermore, heparan sulfate, an IL-2-binding element of cell surfaces and extracellular matrix, or an engineered IL-2 immunocytokine can also direct IL-2 signaling toward this alternative pathway. Overall, these data reveal a non-canonical mechanism for IL-2 signaling that promotes suppressive functions of Tregs, further elucidates how IL-2 supports immune homeostasis, and suggests approaches to promote or suppress Treg functions.

Prediction of creep properties is crucial but still very challenging for the heat-resistant alloys, for which data-driven materials genome technologies provide a promising approach. To this end, we have explored the application of deep learning to quantitatively predict the creep rupture time of Fe–Cr–Ni heat-resistant alloys using textual (chemical composition and creep testing conditions) and visual (as-cast microstructure) data. The multi-layer perceptron (MLP) and the convolutional neural network (CNN) are adopted to analyze the textual and visual data, respectively. Then the multi-source heterogeneous data are integrated by the fusion deep learning model, which achieves a significant enhancement in prediction accuracy compared with the single MLP and CNN models. The results demonstrate that the fusion model is efficient in leveraging information from diverse types of data and learning the relation between various features and creep properties in a high-throughput, statistically robust, and physically meaningful manner. The developed model can be regarded as a new workflow to predict creep properties and give insights into the inverse design of alloys. Our work is equally applicable to other classes of materials and properties as well, where the accumulated data can be reconsidered to guide discoveries.

Extracellular vesicles (EVs) are increasingly recognized as important intermediaries of intercellular communication. They have significant roles in many physiological and pathological processes and show great promise as novel biomarkers of disease, therapeutic agents, and drug delivery tools. Existing studies have shown that natural killer cell-derived EVs (NEVs) can directly kill tumor cells and participate in the crosstalk of immune cells in the tumor microenvironment. NEVs own identical cytotoxic proteins, cytotoxic receptors, and cytokines as NK cells, which is the biological basis for their application in antitumor therapy. The nanoscale size and natural targeting property of NEVs enable precisely killing tumor cells. Moreover, endowing NEVs with a variety of fascinating capabilities via common engineering strategies has become a crucial direction for future research. Thus, here we provide a brief overview of the characteristics and physiological functions of the various types of NEVs, focusing on their production, isolation, functional characterization, and engineering strategies for their promising application as a cell-free modality for tumor immunotherapy.

Abstract A high-precision evaluation of ultrasonic detection sensitivity for a micro-crack can be restricted by a corroded rough surface when the surface microtopography is of the same order of magnitude as the crack depth. In this study, a back-surface micro-crack is considered as a research target. A roughness-modified ultrasonic testing model for micro-cracks is established based on a multi-Gaussian beam model and the principle of phase-screen approximation. The echo signals of micro-cracks and noises corresponding to different rough front surfaces and rough back surfaces are obtained based on a reference reflector signal acquired from a two-dimensional simulation model. Further comparison between the analytical and numerical models shows that the responses of micro-cracks under the effects of different corroded rough surfaces can be accurately predicted. The numerical and analytical results show that the echo signal amplitude of the micro-crack decreases significantly with an increase in roughness, whereas the noise amplitude slightly increases. Moreover, the effect of the rough front surface on the echo signal of the micro-crack is greater than that of the rough back surface. When the root-mean-square (RMS) height of the surface microtopography is less than 15 μm, the two rough surfaces have less influence on the echo signals detected by a focused transducer with a frequency of 5 MHz and diameter of 6 mm. A method for predicting and evaluating the detection accuracy of micro-cracks under different rough surfaces is proposed by combining the theoretical model and a finite element simulation. Then, a series of rough surface samples containing different micro-cracks are fabricated to experimentally validate the evaluation method.

Neutrophils are the most abundant leukocyte in human blood. They are critical for fighting infections and are involved in inflammatory diseases. Mitochondria are indispensable for eukaryotic cells, as they control the biochemical processes of respiration and energy production. Mitochondria in neutrophils have been underestimated since glycolysis is a major metabolic pathway for fuel production in neutrophils. However, several studies have shown that mitochondria are greatly involved in multiple neutrophil functions as well as neutrophil-related diseases. In this review, we focus on how mitochondrial components, metabolism, and related genes regulate neutrophil functions and relevant diseases.

Aircraft cloud seeding refers to the use of equipment on aircraft to release chemicals into clouds, changing their physical and chemical properties to increase rainfall or snowfall. The purpose of precipitation enhancement is to alleviate drought and water scarcity issues. Due to the complexity of the technology, the precise control of factors such as cloud characteristics and chemical release amounts is necessary. Therefore, a scientific evaluation of the potential of aircraft cloud seeding can help to improve the effectiveness of the process, and is currently a technical challenge in weather modification. This study used the mesoscale numerical model WRF coupled with a catalytic process to simulate and evaluate the seven aircraft cloud seeding operations conducted in Hunan Province in 2021. The results show that WRF can effectively evaluate the effectiveness of cloud seeding. When the water vapor conditions are suitable, the airborne dispersion of silver iodide (AgI) can significantly increase the content of large particles of high-altitude ice crystals, snow, and graupel, resulting in an increase in low-level rainwater content and, correspondingly, an increase in ground precipitation. When the water vapor conditions are insufficient, the dispersion of AgI does not trigger effective precipitation, consistent with the results of station observations and actual flight evaluations. This study provides an effective method for scientifically evaluating the potential and effectiveness of aircraft cloud seeding operations.

Abstract Background Critically ill patients diagnosed with COVID-19 may develop a pro-thrombotic state that places them at a dramatically increased lethal risk. Although platelet activation is critical for thrombosis and is responsible for the thrombotic events and cardiovascular complications, the role of platelets in the pathogenesis of COVID-19 remains unclear. Methods Using platelets from healthy volunteers, non-COVID-19 and COVID-19 patients, as well as wild-type and hACE2 transgenic mice, we evaluated the changes in platelet and coagulation parameters in COVID-19 patients. We investigated ACE2 expression and direct effect of SARS-CoV-2 virus on platelets by RT-PCR, flow cytometry, Western blot, immunofluorescence, and platelet functional studies in vitro, FeCl3-induced thrombus formation in vivo, and thrombus formation under flow conditions ex vivo. Results We demonstrated that COVID-19 patients present with increased mean platelet volume (MPV) and platelet hyperactivity, which correlated with a decrease in overall platelet count. Detectable SARS-CoV-2 RNA in the blood stream was associated with platelet hyperactivity in critically ill patients. Platelets expressed ACE2, a host cell receptor for SARS-CoV-2, and TMPRSS2, a serine protease for Spike protein priming. SARS-CoV-2 and its Spike protein directly enhanced platelet activation such as platelet aggregation, PAC-1 binding, CD62P expression, α granule secretion, dense granule release, platelet spreading, and clot retraction in vitro, and thereby Spike protein enhanced thrombosis formation in wild-type mice transfused with hACE2 transgenic platelets, but this was not observed in animals transfused with wild-type platelets in vivo. Further, we provided evidence suggesting that the MAPK pathway, downstream of ACE2, mediates the potentiating role of SARS-CoV-2 on platelet activation, and that platelet ACE2 expression decreases following SARS-COV-2 stimulation. SARS-CoV-2 and its Spike protein directly stimulated platelets to facilitate the release of coagulation factors, the secretion of inflammatory factors, and the formation of leukocyte–platelet aggregates. Recombinant human ACE2 protein and anti-Spike monoclonal antibody could inhibit SARS-CoV-2 Spike protein-induced platelet activation. Conclusions Our findings uncovered a novel function of SARS-CoV-2 on platelet activation via binding of Spike to ACE2. SARS-CoV-2-induced platelet activation may participate in thrombus formation and inflammatory responses in COVID-19 patients.

The ultrasonic detectability of buried defects within composite materials is dependent on the anisotropy of the composite material by which the propagation property of acoustic wave in each direction is variably affected. In this study, the characteristics of acoustic waves propagating in different directions for composite materials are explored based on the full matrix capture (FMC) data using an ultrasonic phased array. The elastic constant of multidirectional carbon fiber reinforced plastic (CFRP) laminate is first derived based on the genetic algorithm. The characteristics of transmitted and reflected waves in higher angles are predicted by implementing the Christoffel equation, and the focal law used in post-processing of FMC data can be optimized accordingly. The imaging results of the total focusing method (TFM) using the improved focal law are compared with the results of the conventional TFM. The results suggest that the optimized TFM can effectively characterize the defect by reducing the background noise. Furthermore, since it is impractical to theoretically correct angle-dependent velocity for in situ inspection, a linear extrapolation method based on the experimentally measurable velocity at low angles is proposed to estimate the velocity profile at higher angles. The imaging results using the fast extrapolated velocity profile is then compared with the theoretical, and it has been demonstrated that while the difference between the images using the theoretical focal law and the linearly extrapolated one is barely visible, the later one is overwhelmingly advantageous to be realiszd for engineering practices.

ABSTRACT Cherubism (CBM), characterized by expansile jawbones with multilocular fibrocystic lesions, is caused by gain‐of‐function mutations in SH3 domain‐binding protein 2 (SH3BP2; mouse orthologue Sh3bp2). Loss of jawbone and dental integrity significantly decrease the quality of life for affected children. Treatment for CBM is limited to multiple surgeries to correct facial deformities. Despite significant advances made with CBM knockin (KI) mouse models (Sh3bp2KI/KI), the activation mechanisms of CBM lesions remain unknown because mutant mice do not spontaneously develop expansile jawbones. We hypothesize that bony inflammation of an unknown cause triggers jawbone expansion in CBM. To introduce jawbone inflammation in a spatiotemporally controlled manner, we exposed pulp of the first right mandibular molar of 6‐week‐old Sh3bp2+/+, Sh3bp2KI/+, and Sh3bp2KI/KI mice. Bacterial invasion from the exposed pulp into root canals led to apical periodontitis in wild‐type and mutant mice. The pathogen‐associated molecular patterns (PAMPs)‐induced inflammation of alveolar bone resulted in jawbone expansion in Sh3bp2KI/+ and Sh3bp2KI/KI mice. CBM‐like lesions developed exacerbated inflammation with increased neutrophil, macrophage, and osteoclast numbers. These lesions displayed excessive neutrophil extracellular traps (NETs) compared to Sh3bp2+/+ mice. Expression levels of IL‐1β, IL‐6, and TNF‐α were increased in periapical lesions of Sh3bp2+/+, Sh3bp2KI/+, and Sh3bp2KI/KI mice and also in plasma and the left untreated mandibles (with no pulp exposure) of Sh3bp2KI/KI mice, suggesting a systemic upregulation. Ablation of Tlr2/4 signaling or depletion of neutrophils by Ly6G antibodies ameliorated jawbone expansion induced by PAMPs in Sh3bp2KI/KI mice. In summary, successful induction of CBM‐like lesions in jaws of CBM mice is important for studying initiating mechanisms of CBM and for testing potential therapies. Our findings further emphasize a critical role of host immunity in the development of apical periodontitis and the importance of maintaining oral health in CBM patients. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

The defects in the welds of energy pipelines have significantly influenced their safe operation. The inefficient and inaccurate detection of the defects may give rise to catastrophic accidents. Ultrasonic phased array inspection is an important means of ensuring pipeline safety. The total focusing method (TFM), using ultrasonic phased arrays, has become widely used in recent years in non-destructive evaluation (NDE). However, manual defect recognition of TFM images is seen to lack accuracy and robustness, arising from deficiency of practical experience. In this paper, the automated classification of different defects from TFM images is studied with a view to facilitate inspection efficacy. By experimentally implementing the TFM approach on a bespoke specimen, the images corresponding to crack-like defects and pore-like defects were employed to investigate the effectiveness of four different machine learning models (known as Support Vector Machine, CART Decision tree, K Nearest Neighbors, Naive Bayes) containing data augmentation, feature extraction and defect classification. The results suggested that the accuracy of defect classification using the HOG-Poly-SVM algorithm was 93%, which outperformed the results from other algorithms. The advantage of the HOG-Poly-SVM algorithm used in defect classification of ultrasonic phased array TFM data is discussed by conducting ten-fold cross validation and other evaluation metrics. In this paper, in order to improve the efficiency of detecting pipeline defects in the future, and for testing test blocks simulating buried pipelines containing defects, we proposed, for the first time, that ultrasonic phased-array TFM imaging results in small object detection images, and found that the SVM algorithm was applicable to ultrasonic phased array TFM imaging, providing a research method and ideas for the use of artificial intelligence in industrial non-destructive testing.

Phagocytes, which include neutrophils, monocytes, macrophages, and dendritic cells, protect the body by removing foreign particles, bacteria, and dead or dying cells. Phagocytic integrins are greatly involved in the recognition of and adhesion to specific antigens on cells and pathogens during phagocytosis as well as the recruitment of immune cells. β2 integrins, including αLβ2, αMβ2, αXβ2, and αDβ2, are the major integrins presented on the phagocyte surface. The activation of β2 integrins is essential to the recruitment and phagocytic function of these phagocytes and is critical for the regulation of inflammation and immune defense. However, aberrant activation of β2 integrins aggravates auto-immune diseases, such as psoriasis, arthritis, and multiple sclerosis, and facilitates tumor metastasis, making them double-edged swords as candidates for therapeutic intervention. Therefore, precise regulation of phagocyte activities by targeting β2 integrins should promote their host defense functions with minimal side effects on other cells. Here, we reviewed advances in the regulatory mechanisms underlying β2 integrin inside-out signaling, as well as the roles of β2 integrin activation in phagocyte functions.

Neutrophils are the most abundant leukocytes in humans and are critical for innate immunity and inflammation. Integrins are critical for neutrophil functions, especially for their recruitment to sites of inflammation or infections. Integrin conformational changes during activation have been heavily investigated but are still not fully understood. Many regulators, such as talin, Rap1-interacting adaptor molecule (RIAM), Rap1, and kindlin, are critical for integrin activation and might be potential targets for integrin-regulating drugs in treating inflammatory diseases. In this review, we outline integrin activation regulators in neutrophils with a focus on the above critical regulators, as well as newly discovered modulators that are involved in integrin activation.

It is a well-documented event that fibroblast growth factors (FGFs) regulate liver development and homeostasis in autocrine, paracrine, and endocrine manners via binding and activating FGF receptors (FGFRs) tyrosine kinase in hepatocytes. Recent research reveals that hepatic stellate cells (HSCs) play a fundamental role in liver immunology. However, how FGF signaling in HSCs regulates liver inflammation remains unclear. Here, we report that FGF promoted NF-κB signaling, an inflammatory pathway, in human HSCs, which was associated with FGFR1 expression. Both FGF and NF-κB signaling in HSCs were compromised by FGFR1 tyrosine kinase inhibitor. After stimulating HSCs with proinflammatory cytokines, expression of multiple FGF ligands was significantly increased. However, disruption of FGF signaling with FGFR inhibitors prominently reduced the apoptosis, inflammatory response, NF-κB nuclear translocation, and expression of matrix metalloproteinase-9 (MMP-9) induced by TNFα in HSCs. Interestingly, FGF21 significantly alleviated the inflammation responses in the concanavalin A (Con A)-induced acutely injured liver. Unlike canonic FGFs that elicit signals through activating the FGFR–heparan sulfate complex, FGF21 activates the FGFR–KLB complex and elicits a different set of signals. Therefore, the finding here indicates the urgency of developing pathway-specific inhibitors that only suppress canonical FGF, but not non-canonical FGF21, signaling for alleviating inflammation in the liver, which is presented in all stages of diseased liver.

Summary: This protocol introduces the SuperSTORM technique, combining stochastic optical reconstruction microscopy (STORM) and molecular modeling. SuperSTORM is optimized for acquiring and processing STORM images of neutrophil integrins but can be used for any cell-surface molecule with known structure and antibody-binding site(s). SuperSTORM identifies molecular cut-offs for eliminating multiple blinks of STORM imaging, determines colocalization, identifies clusters, and reveals molecular orientations and distributions. This protocol extends STORM imaging to cells in microfluidic systems. Improved resolution is achieved by using biomolecule-inherent parameters.For complete information on the generation and use of this protocol, please refer to the paper by Fan et al. (2019).

Strain-controlled continuous fatigue and creep–fatigue tests were carried out at 700 °C and 800 °C on Inconel 625 alloy. The effects of strain rate and tensile-hold time on cyclic stress response and fatigue life were investigated. Then, the microstructural analysis and the fractographic analysis of fatigue-fractured specimens were performed by scanning electron microscopy and transmission electron microscopy. The cyclic stress responses during high-temperature fatigue and the creep–fatigue–oxidation damage mechanism were discussed. The results showed that the strain rate and the tensile-hold time had little effect on the fatigue life at 700 °C, but there was a significant impact at 800 °C due to the creep–fatigue–oxidation interaction. The cyclic plastic deformation accelerated the precipitation of the γ″ phase, resulting in a continuous cyclic hardening and negative strain rate sensitivity. The fatigue failures at 700 °C under continuous fatigue conditions occurred with a transgranular fracture mode, while a transgranular-intergranular hybrid fracture manner was found at 800 °C. Furthermore, a frequency-modified total strain energy density model was proposed to consider the effects of creep and oxidation on fatigue life, and the predicted fatigue lives were located within the 1.5 times error band.

Integrin-based therapeutics could block inflammatory processes but they also impair host defence, limiting their usefulness. Here the authors report an anti-Mac1 antibody that blocks its interaction with pro-inflammatory ligand CD40L but not other ligands, and show that it can protect against sepsis in mice.

The adaptive immune response involves T cell differentiation and migration to sites of inflammation. T cell trafficking is initiated by rolling on inflamed endothelium. Tethers and slings, discovered in neutrophils, facilitate cell rolling at high shear stress. Here, we demonstrate that the ability to form tethers and slings during rolling is highly inducible in T helper 1 (Th1), Th17, and regulatory T (Treg) cells but less in Th2 cells. In vivo, endogenous Treg cells rolled stably in cremaster venules at physiological shear stress. Quantitative dynamic footprinting nanoscopy of Th1, Th17, and Treg cells uncovered the formation of multiple tethers per cell. Human Th1 cells also showed tethers and slings. RNA sequencing (RNA-seq) revealed the induction of cell migration and cytoskeletal genes in sling-forming cells. We conclude that differentiated CD4 T cells stabilize rolling by inducible tether and sling formation. These phenotypic changes approximate the adhesion phenotype of neutrophils and support CD4 T cell access to sites of inflammation.

Abstract Coumarin and its derivatives are widely used as fragrances in industrial products and have medical value. The goal of the present study was to discover genes and pathways related to coumarin biosynthesis in Melilotus albus using transcriptome analysis. The genes of five M. albus near-isogenic lines (NILs) that had different coumarin content and β-glucosidase activity according to the investigation of pedigree were quantified and then analysed by RNA-Seq. Using transcriptome analysis, differentially expressed genes (DEGs) were identified in two pairwise comparisons that differed in coumarin content as well as in two pairwise comparisons that differed in β-glucosidase activity. Gene expression pattern analysis suggested similar transcriptional trends in the genotypes with the same coumarin levels. Furthermore, the Kyoto Encyclopedia of Genes and Genomes (KEGG) database of DEGs was used to identify functional pathways associated with coumarin biosynthesis. We identified 111 unigenes, with several DEGs among them possibly being related to coumarin synthesis pathways. Unigenes encoding a hexokinase, an abscisic acid receptor, a phenylalanine ammonia-lyase (PAL) and two peroxidases particularly showed correspondence with the coumarin content of different genotypes. These results will contribute to a better understanding of the coumarin biosynthesis in M. albus.

Integrin β2 attachment regulates inflammation via effects on neutrophil rolling and extravasation through sequential integrin extension then headpiece opening. Here the authors show an alternative open headpiece prior to extension stabilized in cisby ICAM-1 that limits neutrophil adhesion.

Soluble CD83 (sCD83) is the extracellular domain of the membrane-bound CD83 molecule, and known for its immunoregulatory functions. Whether and how sCD83 participates in the pathogenesis of uveitis, a serious inflammatory disease of the eye that can cause visual disability and blindness, is unknown. By flow cytometry and imaging studies, we show that sCD83 alleviates experimental autoimmune uveitis (EAU) through a novel mechanism. During onset and recovery of EAU, the level of sCD83 rises in the serum and aqueous humor, and CD83+ leukocytes infiltrate the inflamed eye. Systemic or topical application of sCD83 exerts a protective effect by decreasing inflammatory cytokine expression, reducing ocular and splenic leukocyte including CD4+ T cells and dendritic cells (DCs). Mechanistically, sCD83 induces tolerogenic DCs by decreasing the synaptic expression of co-stimulatory molecules and hampering the calcium response in DCs. These changes are caused by a disruption of the cytoskeletal rearrangements at the DC–T cell contact zone, leading to altered localization of calcium microdomains and suppressed T-cell activation. Thus, the ability of sCD83 to modulate DC-mediated inflammation in the eye could be harnessed to develop new immunosuppressive therapeutics for autoimmune uveitis.

T-cell activation requires the formation of the immunological synapse (IS) between a T-cell and an antigen-presenting cell (APC) to control the development of the adaptive immune response. However, calcium release, an initial signal of T-cell activation, has been found to occur before IS formation. The mechanism for triggering the calcium signaling and relationship between calcium release and IS formation remains unclear. Herein, using live-cell imaging, we found that intercellular adhesion molecule 1 (ICAM-1), an essential molecule for IS formation, accumulated and then was depleted at the center of the synapse before complete IS formation. During the process of ICAM-1 depletion, calcium was released. If ICAM-1 failed to be depleted from the center of the synapse, the sustained calcium signaling could not be induced. Moreover, depletion of ICAM-1 in ISs preferentially occurred with the contact of antigen-specific T-cells and dendritic cells (DCs). Blocking the binding of ICAM-1 and lymphocyte function-associated antigen 1 (LFA-1), ICAM-1 failed to deplete at the center of the synapse, and calcium release in T-cells decreased. In studying the mechanism of how the depletion of ICAM-1 could influence calcium release in T-cells, we found that the movement of ICAM-1 was associated with the localization of LFA-1 in the IS, which affected the localization of calcium microdomains, ORAI1 and mitochondria in IS. Therefore, the depletion of ICAM-1 in the center of the synapse is an important factor for an initial sustained calcium release in T-cells.

Mass sensitivity plays a crucial role in the practical application of quartz crystal microbalances (QCMs)-based quantitative analysis. n-m type QCMs have many applications, so it is necessary to clarify the relationship between the mass sensitivity and the electrode of the n-m type QCM. The performance of gold-plated films with different electrodes was studied by theoretical calculation and experiment. The results show that the mass sensitivity on the surface of the n electrode and the surface of the m electrode are essentially the same. Meanwhile, the mass sensitivity of n-m type QCMs varies with the diameter of the n and m electrodes. When the diameter of the n electrode is close to half the diameter of the m electrode, mass sensitivity is at maximum value. These results are important for the further designs and applications of n-m type QCMs.

Photodynamic therapy (PDT) has been commonly used in treating many diseases, such as cancer and infectious diseases. We investigated the different effects of PDT on three main pathogenic bacteria of periodontitis — Prevotella melaninogenica (P.m.), Porphyromonas gingivalis (P.g.) and Aggregatibacter actinomycetemcomitans (A.a.). The portable red light-emitting diode (LED) phototherapy device was used to assess the exogenous PDT effects with different light doses and photosensitizer concentrations (Toluidine blue O, TBO). The portable blue LED phototherapy device was used to assess the endogenous PDT effects with the use of endogenous photosensitizers (porphyrin) under different light doses. We found out that both exogenous and endogenous PDT were able to restrict the growth of all the three bacteria significantly. Moreover, the optimal PDT conditions for these bacteria were obtained through this in vitro screening and could guide the clinical PDT on periodontitis.

Immune checkpoint regulation is a negative feedback regulatory mechanism in the body, and sequential death receptor-1 (PD-1) and programmed death receptor ligand-1 (PD-L1) are known as immune checkpoints. The PD-1/PD-L1 pathway inhibits the activity of effector T cells through a negative regulatory mechanism to avoid excessive response-induced body damage. PD-L1 is highly expressed in many tumor tissues, and high PD-L1 expression can ultimately lead to tumor immune escape. Therefore, immune checkpoint blockade with inhibition of negative immune regulation therapy has become a cutting-edge hot spot for antitumor therapy, with the main target molecules being PD-1 and PD-L1. Recent years have seen promising progress in the study of traditional Chinese medicines and their effects on gastric and colon cancers, particularly in relation to the PD-1/PD-L1 pathway mechanisms. This review specifically examines the modulation of the PD-L1 pathway by certain traditional Chinese medicines in gastric and colon cancers, aiming to provide insights for the development of innovative drugs for these types of digestive cancers. [ABSTRACT FROM AUTHOR]

Warming radiative forced effect (RFE) derived from atmospheric amines attracts lots of attentions because of their contributions to brown carbons. Herein, the enhanced influence of amines (methyl-, dimethyl-, and trimethylamine) on cooling RFE of mineral particles is first confirmed at visible wavelengths. Present results state heterogeneous uptake and oxidation reactions of atmospheric amines are feasible on mineral particle at clean/polluted conditions, which are proofed by related thermodynamics and kinetics data obtained using combined classical molecular dynamics and density function theory methods. Based on mineral particles, simple forcing efficiency (SFE) results explain that amine uptake induces at least 11.8% - 29.5% enhancement on cooling RFE of amine-mineral particles at visible wavelengths. After amines' heterogeneous oxidation, oxidized amine-mineral particles' cooling RFE are furthermore enhanced due to increased oxygen contents. Moreover, oxidized trimethylamine-mineral particle under clean condition shows 45.6% - 47.1% SFE increment at 400-600 nm, which is at least 13.5% higher than that of itself under polluted condition, due to high-oxygen-content product formation through trimethylamine autoxidation. Our results suggest cooling RFE derived from atmospheric amines can be equally important to their warming RFE on atmosphere. It is necessary to update heterogeneous oxidation mechanism and kinetics data of amines in atmospheric model in order to accurately evaluate the whole RFE caused by amines on atmosphere. [ABSTRACT FROM AUTHOR]