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The fundamental communication problem in the wireless Internet of Things (IoT) is to discover a massive number of devices and to allow them reliable access to shared channels. Oftentimes these devices transmit short messages randomly and sporadically. This paper proposes a novel signaling scheme for grant-free massive access, where each device encodes its identity and/or information in a sparse set of tones. Such transmissions are implemented in the form of orthogonal frequency-division multiple access (OFDMA). Under some mild conditions and assuming device delays to be bounded unknown multiples of symbol intervals, sparse OFDMA is proved to enable arbitrarily reliable asynchronous device identification and message decoding with a codelength that is O(K(log K + log S + log N)), where N denotes the device population, K denotes the actual number of active devices, and log S is essentially equal to the number of bits a device can send (including its identity). By exploiting the Fast Fourier Transform (FFT), the computational complexity for discovery and decoding can be made to be sub-linear in the total device population. To prove the concept, a specific design is proposed to identify up to 100 active devices out of $2^{38}$ possible devices with up to 20 symbols of delay and moderate signal-to-noise ratios and fading. The codelength compares much more favorably with those of standard slotted ALOHA and carrier-sensing multiple access (CSMA) schemes.

Lesion mimic mutants (LMMs) are valuable genetic resources for unraveling plant defense responses including programmed cell death. Here, we identified a rice (Oryza sativa) LMM, spotted leaf 38 (spl38), and demonstrated that spl38 is essential for the formation of hypersensitive response-like lesions and innate immunity. Map-based cloning revealed that SPL38 encodes MEDIATOR SUBUNIT 16 (OsMED16). The spl38 mutant showed enhanced resistance to rice pathogens Magnaporthe oryzae and Xanthomonas oryzae pv. oryzae (Xoo) and exhibited delayed flowering, while OsMED16-overexpressing plants showed increased rice susceptibility to M. oryzae. The OsMED16-edited rice lines were phenotypically similar to the spl38 mutant but were extremely weak, exhibited growth retardation, and eventually died. The C-terminus of OsMED16 showed interaction with the positive immune regulator PATHOGENESIS RELATED 3 (OsPR3), resulting in the competitive repression of its chitinase and chitin-binding activities. Furthermore, the ospr3 osmed16 double mutants did not exhibit the lesion mimic phenotype of the spl38 mutant. Strikingly, OsMED16 exhibited an opposite function in plant defense relative to that of Arabidopsis (Arabidopsis thaliana) AtMED16, most likely because of 2 amino acid substitutions between the monocot and dicot MED16s tested. Collectively, our findings suggest that OsMED16 negatively regulates cell death and immunity in rice, probably via the OsPR3-mediated chitin signaling pathway.

Acute myeloid leukemia (AML) is fueled by leukemic stem cells (LSC) whose determinants are challenging to discern from hematopoietic stem cells (HSC) or uncover by approaches focused on general cell properties. We have identified a set of RNA-binding proteins (RBP) selectively enriched in human AML LSCs. Using an in vivo two-step CRISPR-Cas9 screen to assay stem cell functionality, we found 32 RBPs essential for LSCs in MLL-AF9;NrasG12D AML. Loss-of-function approaches targeting key hit RBP ELAVL1 compromised LSC-driven in vivo leukemic reconstitution, and selectively depleted primitive malignant versus healthy cells. Integrative multiomics revealed differentiation, splicing, and mitochondrial metabolism as key features defining the leukemic ELAVL1–mRNA interactome with mitochondrial import protein, TOMM34, being a direct ELAVL1-stabilized target whose repression impairs AML propagation. Altogether, using a stem cell–adapted in vivo CRISPR screen, this work demonstrates pervasive reliance on RBPs as regulators of LSCs and highlights their potential as therapeutic targets in AML. Significance: LSC-targeted therapies remain a significant unmet need in AML. We developed a stem-cell–adapted in vivo CRISPR screen to identify key LSC drivers. We uncover widespread RNA-binding protein dependencies in LSCs, including ELAVL1, which we identify as a novel therapeutic vulnerability through its regulation of mitochondrial metabolism. This article is highlighted in the In This Issue feature, p. 171

Aims This study aimed to provide a safe, stable and efficient SARS-CoV-2 oral vaccine development strategy based on the type III secretion system of attenuated Salmonella and a reference for the development of a SARS-CoV-2 vaccine. Methods and Results The attenuated Salmonella mutant ΔhtrA-VNP was used as a vector to secrete the antigen SARS-CoV-2 based on the type III secretion system (T3SS). The Salmonella pathogenicity island 2 (SPI-2)-encoded T3SS promoter (sifB) was screened to express heterologous antigens (RBD, NTD, S2), and the SPI-2-encoded secretion system (sseJ) was employed to secrete this molecule (psifB-sseJ-antigen, abbreviated BJ-antigen). Both immunoblotting and fluorescence microscopy revealed effective expression and secretion of the antigen into the cytosol of macrophages in vitro. The mixture of the three strains (BJ-RBD/NTD/S2, named AisVax) elicited a marked increase in the induction of IgA or IgG S-protein Abs after oral gavage, intraperitoneal and subcutaneous administration. Flow cytometric analysis proved that AisVax caused T-cell activation, as shown by a significant increase in CD44 and CD69 expression. Significant production of IgA or IgG N-protein Abs was also detected by using psifB-sseJ-N(FL), indicating the universality of this strategy. Conclusions Delivery of multiple SARS-CoV-2 antigens using the type III secretion system of attenuated Salmonella ΔhtrA-VNP is a potential COVID-19 vaccine strategy. Significance and Impact of the Study The attenuated Salmonella strain ΔhtrA-VNP showed excellent performance as a vaccine vector. The Salmonella SPI-2-encoded T3SS showed highly efficient delivery of SARS-COV-2 antigens. Anti-loss elements integrated into the plasmid stabilized the phenotype of the vaccine strain. Mixed administration of antigen-expressing strains improved antibody induction.

It is a main direction of industrial application that manipulator performs grasping task in task space and combines obstacle avoidance planning.It is a main direction of industrial application that manipulator performs grasping task in task space and combines obstacle avoidance planning.I This paper presents an improved RRTConnect path planning algorithm IRRTC based on dynamic constrained space and admittance control. The sliding constraints on the Cartesian position space is used to search for the reachable path of the traction robot by the algorithm. For orientation control, a priori path is formed by artificially given constraint interval, and then the dynamic admittance control is modified by using the end grasping load moment estimated by the joint torque sensor of the manipulator as the constraint. At the same time, the algorithm designs a local escape algorithm based on the current state and target state of the robotic arm, which overcomes the problem that the traditional RRTConnect and RRT cannot escape the local minimum. The simulation and experimental results of the Xmate3 redundant manipulator show that a feasible path can be quickly searched based on constraints and local escape strategies, and a relatively optimal trajectory can be planned based on the original path by the optimized path and also meet the constraints. Within 3S, the average planning success rate is increased to over 90%, and without time constraint, the success rate is 100%, the efficiency is higher than other algorithms. Because the admittance control is used in conjunction with the joint torque feedback to maintain the dynamic correction of the end orientation, which improves the stability of the manipulator. The algorithm can be artificially designed to solve the obstacle avoidance and task constraints in disorderly grasping.

There is an urgent need for animal models of coronavirus disease 2019 to study immunopathogenesis and test therapeutic intervenes. In this study, we showed that NOD/SCID IL2rg

Compared to single functional materials, multifunctional materials with magnetism and luminescence are more attractive and promising; Thus, it has become an important subject. In our work, bifunctional Fe3O4/Tb(acac)3phen/polystyrene) microfibers with magnetic and luminescent properties (acac: acetylacetone, phen: 1,10-phenanthroline) were synthesized by simple electrospinning process. The doping of Fe3O4 and Tb(acac)3phen made the fiber diameter larger. The surface of pure polystyrene microfibers and microfibers doped only with Fe3O4 nanoparticles were chapped similar to bark, whereas the surface of the microfibers was smoother after doping with Tb(acac)3phen complexes. The luminescent properties of the composite microfibers were systematically studied in contrast to pure Tb(acac)3phen complexes, including excitation and emission spectra, fluorescence dynamics, and the temperature dependence of intensity. Compared with the pure complexes, the thermal activation energy and thermal stability of composite microfiber was significantly improved, and the luminescence of the unit mass of Tb(acac)3phen complexes in composite microfibers was stronger than that in pure Tb(acac)3phen complexes. The magnetic properties of the composite microfibers were also investigated using hysteresis loops, and an interesting experimental phenomenon was found that the saturation magnetization of the composite microfibers gradually increased with the increase in the doping proportion of terbium complexes.

To observe the characteristics of suction loss in small-incision lenticule extraction (SMILE) and analyze the factors affecting the stability of the suction ring, classify and grade suction loss, and determine the principles for its prevention and control.This study enrolled patients who underwent SMILE between June 2014 and June 2017. The relationship between the stability of the suction ring and suction loss was ascertained using surgical records and video recordings. The suction loss was classified and graded according to its characteristics and relationship with eye or head movement. The effect of target intervention on suction loss was observed.Suction loss can be divided into sudden and progressive types. According to the severity, the latter was divided into three grades (grade 1A or 1B, 2 and 3). Of the 1200 eyes (608 patients), two (0.17%) had sudden suction loss, and 132 (11%) had progressive suction loss. The superior part, inferior part, and other parts accounted for 63.4%, 19.3%, and 17.3% of progressive suction loss, respectively. The proportion of grades 1A, 1B, 2, and 3 suction loss was 53%, 34%, 12%, and 1%, respectively. The location of the threatened suction loss was opposite to the direction of the patient's head movement. After the intraoperative intervention, grades 1 and 2 did not develop into "actual" suction loss.Progressive suction loss was the most frequently observed suction loss during SMILE procedure. Grading suction loss can elucidate its underlying mechanism, which can guide targeted intervention measures to effectively control and reduce suction loss-induced damage, and further improve the safety and efficacy of SMILE.

The dictionary learning method has been successfully applied to denoise and interpolate seismic data. However, this method cannot be used to adequately interpret weak seismic events and structural features. By combining dictionary learning and a convolutional neural network (CNN) denoiser, we have constructed a new dictionary learning method regularized by a supervised denoiser (DL-SD). In addition to the sparse prior used in previous dictionary learning, the CNN denoiser learns from sizeable amounts of natural images using a deep neural network to help regularize the fine and structural features of data in the DL-SD. We use the plug-and-play alternating directional method of multipliers to solve the net-transform balanced DL-SD model. The results of simultaneous denoising and interpolation indicates that the proposed method is more effective than a deep learning method called the FFDNet and a dictionary learning method known as the data-driven tight frame.

To develop a suitable clinical laboratory assay for detecting the activity of circulating immune complexes (CICs) that activate complement (ACIC).CICs measured in serum were initially used to activate complement, and the remaining complement was activated through sensitized human O-erythrocytes. ACIC was quantified by the degree of hemolysis. Each serum sample was tested for ten consecutive days to determine its stability. Reference ranges are suggested. ACIC was measured in both healthy individuals and patients with autoimmune diseases as well.The OD values of the hemolysis degree index were inversely proportional to ACIC (r = -0.986, P = 0.002). A pooled serum was used to eliminate interference and optimize the experiment. The hemolysis degree (HD) was used to indicate the detection result. HD = (detection value OD/negative value OD)*100. Both the intra-batch and the inter-batch results showed good stability with a CV of 6.5% and 8.1%, respectively. HD differences between males and females were significant (P = 0.015) while the normal distribution for both genders was conformed. The HD recommended reference range for men is 56-88 while for women is 51-86. Serum HD of healthy subjects and autoimmune disease patients showed a significant difference (P = 0.001). Autoimmune disease patients have lower HD which was a result of having stronger ACIC.The ACIC assay while utilizing human O-erythrocytes as an indicator system is sensitive and accurate, and has potential in clinical applications.

Most of the aircraft skin is assembled by riveted structural parts, and the riveted holes are prone to cracks due to stress concentration, so it is urgent to carry out non-destructive testing research on aircraft riveted components. In this paper, a planar remote-field eddy current focusing detection probe was designed to study the hidden defects around the holes of aircraft riveted components, and a three-dimensional simulation model was established for the detection of hidden defects in aircraft riveted components. The structural parameters of the focused remote-field eddy current probe were optimized by combining simulation and experiment, and the planar remote-field eddy current focusing detection test was carried out. The simulation and results showed that: When the inclination angle of the excitation coil is 10° and the coil spacing is 0mm, the optimized focused remote-field eddy current probe can effectively detect the defect in length × width × depth of 10mm×0.2mm×1mm under the buried depth of 9mm. When the buried depth of the defect is 6mm-9mm, the focused remote-field eddy current probe has a better detection ability than the non-focused remote-field eddy current probe.

Background BCMA CAR-T is highly effective for relapsed/refractory multiple myeloma(R/R-MM) and significantly improves the survival of patients. However, the short remission time and high relapse rate of MM patients treated with BCMA CAR-T remain bottlenecks that limit long-term survival. The immune microenvironment of the bone marrow (BM) in R/R-MM may be responsible for this. The present study aims to present an in-depth analysis of resistant mechanisms and to explore potential novel therapeutic targets for relapse of BCMA CAR-T treatment via single-cell RNA sequencing (scRNA-seq) of BM plasma cells and immune cells. Methods This study used 10X Genomic scRNA-seq to identify cell populations in R/R-MM CD45+ BM cells before BCMA CAR-T treatment and relapse after BCMA CAR-T treatment. Cell Ranger pipeline and CellChat were used to perform detailed analysis. Results We compared the heterogeneity of CD45+ BM cells before BCMA CAR-T treatment and relapse after BCMA CAR-T treatment. We found that the proportion of monocytes/macrophages increased, while the percentage of T cells decreased at relapse after BCMA CAR-T treatment. We then reclustered and analyzed the alterations in plasma cells, T cells, NK cells, DCs, neutrophils, and monocytes/macrophages in the BM microenvironment before BCMA CAR-T treatment and relapse after BCMA CAR-T treatment. We show here that the percentage of BCMA positive plasma cells increased at relapse after BCMA CAR-T cell therapy. Other targets such as CD38, CD24, SLAMF7, CD138, and GPRC5D were also found to be expressed in plasma cells of the R/R-MM patient at relapse after BCMA CAR-T cell therapy. Furthermore, exhausted T cells, TIGIT+NK cells, interferon-responsive DCs, and interferon-responsive neutrophils, increased in the R/R-MM patient at relapse after BCMA CAR-T cell treatment. Significantly, the proportion of IL1βhi Mφ, S100A9hi Mφ, interferon-responsive Mφ, CD16hi Mφ, MARCO hi Mφ, and S100A11hi Mφ significantly increased in the R/R-MM patient at relapse after BCMA CAR-T cell therapy. Cell–cell communication analysis indicated that monocytes/macrophages, especially the MIF and APRIL signaling pathway are key players in R/R-MM patient at relapse after BCMA CAR-T cell therapy. Conclusion Taken together, our data extend the understanding of intrinsic and extrinsic relapse of BCMA CAR-T treatment in R/R-MM patient and the potential mechanisms involved in the alterations of antigens and the induced immunosuppressive microenvironment, which may provide a basis for the optimization of BCMA CAR-T strategies. Further studies should be performed to confirm these findings.

Table S5 shows the log2 fold change of differentially expressed transcripts from the ELAVL1 knockout RN2c RNA-sequencing data set.

Table S7 shows (a) reproducible peaks and (b) GO analysis of ELAVL1-bound transcripts identified in the eCLIP-sequencing data set. The negatively enriched (c) and positively-enriched (d) gene sets identified by GSEA of ELAVL1-bound and differentially expressed transcripts from the RN2c RNA-sequencing data set are shown.

Table S1 shows expression of RBPs in an RNA-sequencing data set of human bone marrow hematopoietic stem and progenitor subpopulations.

Supplemental Figure S4 shows the validation of ELAVL1 knockdown by western blot and effects of ELAVL1 loss on human primary AML cells in vitro and in vivo.

Table S8 shows transcripts from the ELAVL1 knockout RNA-sequencing data set that are (a) alternatively spliced and (b) alternatively spliced as well as bound by ELAVL1 as identified by eCLIP-sequencing.

Acute myeloid leukemia (AML) is fueled by leukemic stem cells (LSC) whose determinants are challenging to discern from hematopoietic stem cells (HSC) or uncover by approaches focused on general cell properties. We have identified a set of RNA-binding proteins (RBP) selectively enriched in human AML LSCs. Using an in vivo two-step CRISPR-Cas9 screen to assay stem cell functionality, we found 32 RBPs essential for LSCs in MLL-AF9;NrasG12D AML. Loss-of-function approaches targeting key hit RBP ELAVL1 compromised LSC-driven in vivo leukemic reconstitution, and selectively depleted primitive malignant versus healthy cells. Integrative multiomics revealed differentiation, splicing, and mitochondrial metabolism as key features defining the leukemic ELAVL1–mRNA interactome with mitochondrial import protein, TOMM34, being a direct ELAVL1-stabilized target whose repression impairs AML propagation. Altogether, using a stem cell–adapted in vivo CRISPR screen, this work demonstrates pervasive reliance on RBPs as regulators of LSCs and highlights their potential as therapeutic targets in AML.Significance:LSC-targeted therapies remain a significant unmet need in AML. We developed a stem-cell–adapted in vivo CRISPR screen to identify key LSC drivers. We uncover widespread RNA-binding protein dependencies in LSCs, including ELAVL1, which we identify as a novel therapeutic vulnerability through its regulation of mitochondrial metabolism.This article is highlighted in the In This Issue feature, p. 171

Supplemental Figure S5 shows the effects of DHTS treatment on THP-1 and human primary AML cells in vitro, the effects of MS-444 treatment on human umbilical cord blood in vivo, and validation of ELAVL1 overexpression by western blot.

Table S9 shows the (a) differentially expressed transcripts and (b) altered pathways as identified by GSEA from the ELAVL1 knockdown human primary AML RNA-sequencing data set.

Table S6 shows altered gene sets identified by GSEA of the ELAVL1 knockout RN2c RNA-sequencing data set.

Supplemental Figure S1 shows expression of RBPs in healthy HSPCs and RN2c cells, sgRNA drop out dynamics in primary and secondary transplants, the molecular functions and biological processes associated with the hit RBPs and their gene essentialities.

Supplemental Figure S6 shows the GSEA of ELAVL1 depletion in mouse RN2c and human primary AML cells, ELAVL1 subcellular localization in mouse and human AML, the qRT-PCR of ELAVL1 targets in MS-444-treated RN2c cells, and the integrative analysis of the mouse RNA-seq and eCLIP-seq.

Table S2 shows sequences of (a) sgRNAs from the RBP screen, (b) sequencing primers, (c) shRNAs and human-targeting sgRNAs, (d) eCLIP oligos.

Supplemental Figure S2 shows validation of sgRNA -mediated knockout of hit RBPs, comparison of hit RBP depletion in RN2c in vitro versus in vivo, effects of ELAVL1 loss on human AML cell lines, characteristics of the MLL-AF9 AML mouse model, and ELAVL1 expression in normal hematopoietic cells.

Table S3 shows clinical information for all AML patient specimens used in this study.

Supplemental Figure S3 shows the generation of the bcCML mouse model and the effects of ELAVL1 on normal bone marrow in vivo.