Total Synthesis of Cyclotripeptidic Natural Products Anacine, Aurantiomide C, Polonimides A and C, and Verrucine F.

The total synthesis of cyclotripeptidic natural products possessing a central piperazino[2,1-b]quinazolin-3,6-dione core is described through an original strategy involving the pivotal cyclocondensation of an electrophilic homoserine lactone intermediate. The alkylidene group was spontaneously installed by autoxidation during the cyclocondensation process, while the propionamide side chain was introduced through the nickel-catalyzed aminocarbonylation of a bromoethyl intermediate. This last reaction is unprecedented on such highly functionalized intermediates. Finally, we explored structural modifications and interconversions of the natural products. Overall, this work led to anacine, aurantiomide C, polonimides A and C, and verrucine F.

Research on the Sensing Characteristics of an Integrated Grid-like Sensor Based on a Triboelectric Nanogenerator.

With the development of the integration and miniaturization of sensing devices, the concept of self-sensing devices has been proposed. A motion state is self-sensed via the structure or integration of an actuator in the construction of a sensing unit. This device is then used to capture the perception and measurement of states such as position, displacement, and speed. A triboelectric nanogenerator converts mechanical energy into electrical energy through the coupling effect of contact generation and electrostatic induction, which represents one of the reliable ways through which to realize integrated sensing. In this world, the power generation technology of the TENG is applied to a sensing device. The sensing characteristics of a grid-like TENG are designed and analyzed in freestanding triboelectric mode. Firstly, a relation model of displacement, velocity, voltage, and charge is established. The charge-transfer increment and current amounts are linearly related to the velocity. The open-circuit voltage has a positive relationship with the displacement. The maximum open-circuit voltage and the maximum charge transfer are fixed values, and they are only related to the inherent parameters of a triboelectric nanogenerator. Next, the sensor model is constructed using COMSOL Multiphysics 6.0. The simulation results show that the relationships between output voltage and charge transfer, as well as those between the increments of charge transfer, velocity, and displacement, are consistent with the results derived from the formula. Finally, a performance test of the designed sensor is carried out, and the results are consistent with the theoretical deduction and simulation. After analysis and processing of the output electrical signal by the host computer, it can feedback the frequency and speed value of the measured object. In addition, the output signal is stable, and there is no large fluctuation or attenuation during the 521-s vibration test. Because the working unit of the sensor is thin filmed, it is small in size, easy to integrate, and has no external power supply; moreover, it can be integrated into a device to realize the self-sensing of a motion state.

In situ annealing achieves an ultrafast synthesis of high coercive strontium ferrite foams and beyond.

Strontium ferrite nanostructures have attracted intensive interest recently due to the increasing demand for cost-effective features and good chemical corrosion resistance of magnetic materials, yet the ultrafast synthesis of strontium ferrite with desired coercivity is still experiencing a severe challenge. Herein, porous strontium ferrite foams with a coercivity up to 23.35 kOe were prepared by ultrafast in situ annealing for 1 min based on an auto-combustion strategy. The high coercivity of strontium ferrite benefits from the increasing magnetocrystalline anisotropy caused by the ion substitution and the appropriate grain size close to the critical single-domain size of strontium ferrite. In addition, this ultrafast synthesis can be extended to prepare a series of porous spinel, lanthanide-based perovskites, and their high-entropy counterpart foams. We also demonstrate that this strategy is feasible for preparing biphasic composite oxide foams. Furthermore, this work provides important guidance for the design of porous permanent magnet materials and the efficient preparation of porous oxide foam materials.

High-Throughput, Quantitative Analysis of Peptide-Exchanged MHCI Complexes by Native Mass Spectrometry.

Immune monitoring in cancer immunotherapy involves screening CD8+ T-cell responses against neoantigens, the tumor-specific peptides presented by Major histocompatibility complex Class I (MHCI) on the cell surface. High-throughput immune monitoring requires methods to produce and characterize small quantities of thousands of MHCI-peptide complexes that may be tested for a patient's T-cell response. MHCI synthesis has been achieved using a photocleavable peptide that is exchanged by the neoantigen; however, assays that measure peptide exchange currently disassemble the complex prior to analysis─precluding direct molecular characterization. Here, we use native mass spectrometry (MS) to profile intact recombinant MHCI complexes and directly measure peptide exchange. Coupled with size-exclusion chromatography or capillary-zone electrophoresis, the assay identified all tested human leukocyte antigen (HLA)/peptide combinations in the nanomole to picomole range with minimal run time, reconciling the synthetic and analytical requirements of MHCI-peptide screening with the downstream T-cell assays. We further show that the assay can be "multiplexed" by measuring exchange of multiple peptides simultaneously and also enables calculation of Vc50, a measure of gas-phase stability. Additionally, MHCI complexes were fragmented by top-down sequencing, demonstrating that the intact complex, peptide sequence, and their binding affinity can be determined in a single analysis. This screening tool for MHCI-neoantigen complexes represents a step toward the application of state-of-the-art MS technology in translational settings. Not only is this assay already informing on the viability of immunotherapy in practice, the platform also holds promise to inspire novel MS readouts for increasingly complex biomolecules used in the diagnosis and treatment of disease.

Lightweight Compound Scaling Network for Nasopharyngeal Carcinoma Segmentation from MR Images.

Nasopharyngeal carcinoma (NPC) is a category of tumours with a high incidence in head-and-neck. To treat nasopharyngeal cancer, doctors invariably need to perform focal segmentation. However, manual segmentation is time consuming and laborious for doctors and the existing automatic segmentation methods require large computing resources, which makes some small and medium-sized hospitals unaffordable. To enable small and medium-sized hospitals with limited computational resources to run the model smoothly and improve the accuracy of structure, we propose a new LW-UNet network. The network utilises lightweight modules to form the Compound Scaling Encoder and combines the benefits of UNet to make the model both lightweight and accurate. Our model achieves a high accuracy with a Dice coefficient value of 0.813 with 3.55 M parameters and 7.51 G of FLOPs within 0.1 s (testing time in GPU), which is the best result compared with four other state-of-the-art models.

CAFS: An Attention-Based Co-Segmentation Semi-Supervised Method for Nasopharyngeal Carcinoma Segmentation.

Accurate segmentation of nasopharyngeal carcinoma is essential to its treatment effect. However, there are several challenges in existing deep learning-based segmentation methods. First, the acquisition of labeled data are challenging. Second, the nasopharyngeal carcinoma is similar to the surrounding tissues. Third, the shape of nasopharyngeal carcinoma is complex. These challenges make the segmentation of nasopharyngeal carcinoma difficult. This paper proposes a novel semi-supervised method named CAFS for automatic segmentation of nasopharyngeal carcinoma. CAFS addresses the above challenges through three mechanisms: the teacher-student cooperative segmentation mechanism, the attention mechanism, and the feedback mechanism. CAFS can use only a small amount of labeled nasopharyngeal carcinoma data to segment the cancer region accurately. The average DSC value of CAFS is 0.8723 on the nasopharyngeal carcinoma segmentation task. Moreover, CAFS has outperformed the state-of-the-art nasopharyngeal carcinoma segmentation methods in the comparison experiment. Among the compared state-of-the-art methods, CAFS achieved the highest values of DSC, Jaccard, and precision. In particular, the DSC value of CAFS is 7.42% higher than the highest DSC value in the state-of-the-art methods.

Short-Range Diffusion Enables General Synthesis of Medium-Entropy Alloy Aerogels.

Medium-entropy alloy aerogels (MEAAs) with the advantages of both multimetallic alloys and aerogels are promising new materials in catalytic applications. However, limited by the immiscible behavior of different metals, achieving single-phase MEAAs is still a grand challenge. Herein, a general strategy for preparing ultralight 3D porous MEAAs with the lowest density of 39.3 mg cm-3 among the metal materials is reported, through combining auto-combustion and subsequent low-temperature reduction procedures. The homogenous mixing of precursors at the ionic level makes the short-range diffusion of metal atoms possible to drive the formation of single-phase MEAAs. As a proof of concept in catalysis, as-synthesized Ni50 Co15 Fe30 Cu5 MEAAs exhibit a high mass activity of 1.62 A mg-1 and specific activity of 132.24 mA cm-2 toward methanol oxidation reactions, much higher than those of the low-entropy counterparts. In situ Fourier transform infrared and NMR spectroscopies reveal that MEAAs can enable highly selective conversion of methanol to formate. Most importantly, a methanol-oxidation-assisted MEAAs-based water electrolyzer can achieve a low cell voltage of 1.476 V at 10 mA cm-2 for making value-added formate at the anode and H2 at the cathode, 173 mV lower than that of traditional alkaline water electrolyzers.

Dihuang-Yinzi Alleviates Cognition Deficits via Targeting Energy-Related Metabolism in an Alzheimer Mouse Model as Demonstrated by Integration of Metabolomics and Network Pharmacology.

Energy metabolism disturbance and the consequent reactive oxygen species (ROS) overproduction play a key and pathogenic role in the onset and progression of Alzheimer's disease (AD). Dihuang-Yinzi (DHYZ) is a traditional Chinese herbal prescription clinically applied to treat AD and other neurodegenerative diseases for a long time. However, the systematical metabolic mechanism of DHYZ against AD remains largely unclear. Here we aimed to explore the mechanism of DHYZ in the treatment of AD comprehensively in an in vivo metabolic context by performing metabolomics analysis coupled with network pharmacology study and experimental validation. The network pharmacology was applied to dig out the potential target of DHYZ against AD. The metabolomics analysis based on UPLC-HRMS was carried out to profile the urine of 2× Tg-AD mice treated with DHYZ. By integrating network pharmacology and metabolomics, we found DHYZ could ameliorate 4 key energy-related metabolic pathways, including glycerophospholipid metabolism, nicotinate/nicotinamide metabolism, glycolysis, and tricarboxylic acid cycle. Besides, we identified 5 potential anti-AD targets of DHYZ, including DAO, HIF1A, PARP1, ALDH3B2, and ACHE, and 14 key differential metabolites involved in the 4 key energy-related metabolic pathways. Furthermore, DHYZ depressed the mitochondrial dysfunction and the resultant ROS overproduction through ameliorating glycerophospholipid metabolism disturbance. Thereby DHYZ increased nicotinamide adenine dinucleotide (NAD+) content and promoted glycolysis and tricarboxylic acid (TCA) cycle, and consequently improved oxidative phosphorylation and energy metabolism. In the present study, we provided a novel, comprehensive and systematic insight into investigating the therapeutic efficacy of DHYZ against AD via ameliorating energy-related metabolism.

Tissue inhibitor of metalloproteinases 1 is involved in ROS-mediated inflammation via regulating matrix metalloproteinase 1 expression in the sea cucumber Apostichopus japonicus.

Tissue inhibitors of metalloproteinases (TIMPs) serve as matrix metalloproteinase (MMP) inhibitors in the pathogenesis of inflammatory diseases in vertebrates. We cloned and characterised the TIMP1 gene from Apostichopus japonicus using RACE approaches (designated as AjTIMP1). For Vibrio splendidus-challenged sea cucumbers, the peak expression of AjTIMP1 mRNAs in coelomocytes was detected at 24 h (23.44-fold) and remained at high levels (4.01-fold) until 72 h. Similarly, AjTIMP1 expression was upregulated in primary coelomocytes exposed to 10 µg mL-1 LPS. AjTIMP1 was expressed in all tissues, and the highest expression was observed in the body wall. Functional investigation revealed an imbalance in the ratio of AjMMP1/AjTIMP1 in the skin ulceration syndrome (SUS) diseased group; it was sharply up-regulated to 3.97:1 compared with the healthy group. Furthermore, when AjTIMP1 was knocked down using small interfering RNA (siRNA-KD) to 0.4-fold, AjMMP1 and AjMMP19 were upregulated to 1.99- and 1.85-fold, respectively. AjTIMP1 siRNA-KD can promote ROS production by 26.2%, whereas AjMMP1 siRNA-KD can eliminate the increase in ROS. In inflamed tissues, collagen I and III levels were decreased by 33.1% and 33.6%, respectively, in the AjTIMP1 siRNA group at 24 h AjTIMP1 was involved in the inflammatory response by mediating ROS formation and collagen degradation.

DCNet: Densely Connected Deep Convolutional Encoder-Decoder Network for Nasopharyngeal Carcinoma Segmentation.

Nasopharyngeal Carcinoma segmentation in magnetic resonance imagery (MRI) is vital to radiotherapy. Exact dose delivery hinges on an accurate delineation of the gross tumor volume (GTV). However, the large-scale variation in tumor volume is intractable, and the performance of current models is mostly unsatisfactory with indistinguishable and blurred boundaries of segmentation results of tiny tumor volume. To address the problem, we propose a densely connected deep convolutional network consisting of an encoder network and a corresponding decoder network, which extracts high-level semantic features from different levels and uses low-level spatial features concurrently to obtain fine-grained segmented masks. Skip-connection architecture is involved and modified to propagate spatial information to the decoder network. Preliminary experiments are conducted on 30 patients. Experimental results show our model outperforms all baseline models, with improvements of 4.17%. An ablation study is performed, and the effectiveness of the novel loss function is validated.

AttR2U-Net: A Fully Automated Model for MRI Nasopharyngeal Carcinoma Segmentation Based on Spatial Attention and Residual Recurrent Convolution.

Radiotherapy is an essential method for treating nasopharyngeal carcinoma (NPC), and the segmentation of NPC is a crucial process affecting the treatment. However, manual segmentation of NPC is inefficient. Besides, the segmentation results of different doctors might vary considerably. To improve the efficiency and the consistency of NPC segmentation, we propose a novel AttR2U-Net model which automatically and accurately segments nasopharyngeal carcinoma from MRI images. This model is based on the classic U-Net and incorporates advanced mechanisms such as spatial attention, residual connection, recurrent convolution, and normalization to improve the segmentation performance. Our model features recurrent convolution and residual connections in each layer to improve its ability to extract details. Moreover, spatial attention is fused into the network by skip connections to pinpoint cancer areas more accurately. Our model achieves a DSC value of 0.816 on the NPC segmentation task and obtains the best performance compared with six other state-of-the-art image segmentation models.

Echinacoside Suppresses Amyloidogenesis and Modulates F-actin Remodeling by Targeting the ER Stress Sensor PERK in a Mouse Model of Alzheimer's Disease.

Endoplasmic reticulum stress (ERS) plays a vital and pathogenic role in the onset and progression of Alzheimer's disease (AD). Phosphorylation of PKR-like endoplasmic reticulum kinase (PERK) induced by ERS depresses the interaction between actin-binding protein filamin-A (FLNA) and PERK, which promotes F-actin accumulation and reduces ER-plasma membrane (PM) communication. Echinacoside (ECH), a pharmacologically active component purified from Cistanche tubulosa, exhibits multiple neuroprotective activities, but the effects of ECH on ERS and F-actin remodeling remain elusive. Here, we found ECH could inhibit the phosphorylation of PERK. Firstly ECH can promote PERK-FLNA combination and modulate F-actin remodeling. Secondly, ECH dramatically decreased cerebral Aß production and accumulation by inhibiting the translation of BACE1, and significantly ameliorated memory impairment in 2 × Tg-AD mice. Furthermore, ECH exhibited high affinity to either mouse PERK or human PERK. These findings provide novel insights into the neuroprotective actions of ECH against AD, indicating that ECH is a potential therapeutic agent for halting and preventing the progression of AD.

Structure of the essential inner membrane lipopolysaccharide-PbgA complex.

Lipopolysaccharide (LPS) resides in the outer membrane of Gram-negative bacteria where it is responsible for barrier function1,2. LPS can cause death as a result of septic shock, and its lipid A core is the target of polymyxin antibiotics3,4. Despite the clinical importance of polymyxins and the emergence of multidrug resistant strains5, our understanding of the bacterial factors that regulate LPS biogenesis is incomplete. Here we characterize the inner membrane protein PbgA and report that its depletion attenuates the virulence of Escherichia coli by reducing levels of LPS and outer membrane integrity. In contrast to previous claims that PbgA functions as a cardiolipin transporter6-9, our structural analyses and physiological studies identify a lipid A-binding motif along the periplasmic leaflet of the inner membrane. Synthetic PbgA-derived peptides selectively bind to LPS in vitro and inhibit the growth of diverse Gram-negative bacteria, including polymyxin-resistant strains. Proteomic, genetic and pharmacological experiments uncover a model in which direct periplasmic sensing of LPS by PbgA coordinates the biosynthesis of lipid A by regulating the stability of LpxC, a key cytoplasmic biosynthetic enzyme10-12. In summary, we find that PbgA has an unexpected but essential role in the regulation of LPS biogenesis, presents a new structural basis for the selective recognition of lipids, and provides opportunities for future antibiotic discovery.

Data on charge separation of bispecific and mispaired IgGs using native charge-variant mass spectrometry.

The data supplied in this work are related to the research article entitled "Characterization of Bispecific and Mispaired IgGs by Native Charge-Variant Mass Spectrometry" (Phung et al., 2019). This data article describes a powerful analytical platform using native weak cation exchange chromatography coupled to a high-resolution mass spectrometer, charge variant mass spectrometry (CV-MS), to characterize bispecific and mispaired antibody species. Elution order is investigated through analytical methods and molecular modeling in an effort to understand the intrinsic charge, size and shape differences of these molecules.

TATA-Box Binding Protein O-GlcNAcylation at T114 Regulates Formation of the B-TFIID Complex and Is Critical for Metabolic Gene Regulation.

In eukaryotes, gene expression is performed by three RNA polymerases that are targeted to promoters by molecular complexes. A unique common factor, the TATA-box binding protein (TBP), is thought to serve as a platform to assemble pre-initiation complexes competent for transcription. Here, we describe a novel molecular mechanism of nutrient regulation of gene transcription by dynamic O-GlcNAcylation of TBP. We show that O-GlcNAcylation at T114 of TBP blocks its interaction with BTAF1, hence the formation of the B-TFIID complex, and its dynamic cycling on and off of DNA. Transcriptomic and metabolomic analyses of TBPT114A CRISPR/Cas9-edited cells showed that loss of O-GlcNAcylation at T114 increases TBP binding to BTAF1 and directly impacts expression of 408 genes. Lack of O-GlcNAcylation at T114 is associated with a striking reprogramming of cellular metabolism induced by a profound modification of the transcriptome, leading to gross alterations in lipid storage.

Native Hydrophobic Interaction Chromatography Hyphenated to Mass Spectrometry for Characterization of Monoclonal Antibody Minor Variants.

Conventionally, hydrophobic interaction chromatography (HIC) uses mobile phases with high salt concentration that are not compatible with mass spectrometry (MS). Here we describe development of an HIC method coupled with MS detection (HIC-MS) utilizing an aqueous mobile phase with a low concentration of a volatile salt for characterizing recombinant monoclonal antibody (mAb) post-translational modifications (PTMs). The ability of HIC to separate the oxidation and free thiol variants of the mAbs enables their isolation and rapid characterization of these attributes under native conditions, an important step toward understanding the role they play.

Interface modeling of magnetorheological elastomers subjected to variable working strain.

A magnetorheological elastomer (MRE) is a type of particle-matrix composite material, whose properties depend on the strain to which it is subjected in different applications. This paper proposes an interface model in which the magnetorheological characteristics of an MRE are described in terms of the effect of variable strain on the strength of interfacial bonding between the particles and the matrix. The model can describe the whole process of interface change from a strong interface to a strong-weak mixed interface and then to a weak interface under variable strain. The results indicate that the combined effects of the magnetic flux density, particle content, and strain amplitude are responsible for the magnetorheological performance of the MRE. The maximum value of the shear modulus under large strain is decreased by 0.75 × 105 Pa compared to the value under small strain. This model opens new opportunities for the development of high-performance MREs and MRE-based devices under variable strain conditions.

Elucidating heavy/light chain pairing preferences to facilitate the assembly of bispecific IgG in single cells.

Multiple strategies have been developed to facilitate the efficient production of bispecific IgG (BsIgG) in single host cells. For example, we previously demonstrated near quantitative (≥90%) formation of BsIgG of different species and isotypes by combining 'knob-into-hole' mutations for heavy chain heterodimerization with engineered antigen-binding fragments (Fabs) for preferential cognate heavy/light chain pairing. Surprisingly, in this study we found high yield (>65%) of BsIgG1without Fab engineering to be a common occurrence, i.e., observed for 33 of the 99 different antibody pairs evaluated. Installing charge mutations at both CH1/CL interfaces was sufficient for near quantitative yield (>90%) of BsIgG1 for most (9 of 11) antibody pairs tested with this inherent cognate chain pairing preference. Mechanistically, we demonstrate that a strong cognate pairing preference in one Fab arm can be sufficient for high BsIgG1 yield. These observed chain pairing preferences are apparently driven by variable domain sequences and can result from a few specific residues in the complementarity-determining region (CDR) L3 and H3. Transfer of these CDR residues into other antibodies increased BsIgG1 yield in most cases. Mutational analysis revealed that the disulfide bond between heavy and light chains did not affect the yield of BsIgG1. This study provides some mechanistic understanding of factors contributing to antibody heavy/light chain pairing preference and subsequently contributes to the efficient production of BsIgG in single host cells.

Identification and characterization of an octameric PEG-protein conjugate system for intravitreal long-acting delivery to the back of the eye.

Innovative protein engineering and chemical conjugation technologies have yielded an impressive number of drug candidates in clinical development including >80 antibody drug conjugates, >60 bispecific antibodies, >35 Fc-fusion proteins and >10 immuno-cytokines. Despite these innovations, technological advances are needed to address unmet medical needs with new pharmacological mechanisms. Age-related eye diseases are among the most common causes of blindness and poor vision in the world. Many such diseases affect the back of the eye, where the inaccessibility of the site of action necessitates therapeutic delivery via intravitreal (IVT) injection. Treatments administered via this route typically have vitreal half-lives <10 days in humans, requiring frequent administration. Since IVT injection is burdensome to patients, there exists a strong need to develop therapeutics with prolonged residence time in the eye. We report here a strategy to increase retention of a therapeutic fragment antibody (Fab) in the eye, using an anti-complement factor D Fab previously optimized for ocular delivery. Polyethylene glycol structures, varying in length, geometry and degree of branching, were coupled to the Fab via maleimide-activated termini. A screening strategy was developed to allow for key determinants of ocular half-life to be measured in vitro. After compound selection, a scalable process was established to enable tolerability and pharmacokinetic studies in cynomolgus monkeys, demonstrating an increase in vitreal half-life with no associated adverse events. Further, we show that the technique for compound selection, analytical characterization, and scalable production is general for a range of antibody fragments. The application of the technology has broad impact in across many therapeutic areas with the first major advancement in the treatment of an important ocular disease.

Fabrication, characterization, and magnetic properties of exchange-coupled porous BaFe8Al4O19/Co0.6Zn0.4Fe2O4 nanocomposite magnets.

Fabrication of exchange-coupled nanocomposite magnets has been considered to be the most effective method to achieve the high energy product for advanced permanent magnet applications. In this work, we report a facile auto-combustion synthesis to prepare porous exchange-coupled hard-soft ferrite-based magnetic BaFe8Al4O19-x wt% Co0.6Zn0.4Fe2O4 nanocomposites (where x = 10, 20, 30 and 40), which realize an effective exchange-coupled interaction when the x value is less than 30. Compared with BaFe8Al4O19, the optimized nanocomposite with 20% Co0.6Zn0.4Fe2O4 shows a 70.3% increase in Ms and a 60.4% enhancement in Mr and maintains a high Hc value of 8.8 kOe. The work demonstrates that the auto-combustion synthesis is a promising approach for the fabrication of high-performance ferrite-based permanent magnets.