The canine anti‐tumor necrosis factor‐alpha (TNF‐α) monoclonal antibody is a potential therapeutic option for treating canine arthritis. The current treatments for arthritis in dogs have limitations ...due to side effects, emphasizing the need for safer and more effective therapies. The crystal structure of canine TNF‐α (cTNF‐α) was successfully determined at a resolution of 1.85 Å, and the protein was shown to assemble as a trimer, with high similarity to the functional quaternary structure of human TNF‐α (hTNF‐α). Adalimumab (Humira), a known TNF‐α inhibitor, effectively targets and neutralizes TNF‐α to reduce inflammation and has been used to manage autoimmune conditions such as rheumatoid arthritis. By comparing the structure of cTNF‐α with the complex structure of hTNF‐α and adalimumab‐Fab, the epitope of adalimumab on cTNF‐α was identified. The significant structural similarities of epitopes in cTNF‐α and hTNF‐α indicate the potential of using adalimumab to target cTNF‐α. Therefore, a canine/human chimeric antibody, Humivet‐R1, was created by grafting the variable domain of adalimumab onto a canine antibody framework derived from ranevetmab. Humivet‐R1 exhibits potent neutralizing ability (IC50 = 0.05 nM) and high binding affinity (EC50 = 0.416 nM) to cTNF‐α, comparable to that of adalimumab for both hTNF‐α and cTNF‐α. These results strongly suggest that Humivet‐R1 has the potential to provide effective treatment for canine arthritis with reduced side effects. Here, we propose a structure‐guided antibody design for the use of a chimeric antibody to treat canine inflammatory disease. Our successful development strategy can speed up therapeutic antibody discovery for animals and has the potential to revolutionize veterinary medicine.
Development of specific antiviral agents is an urgent unmet need for SARS-coronavirus 2 (SARS-CoV-2) infection. This study focuses on host proteases that proteolytically activate the SARS-CoV-2 spike ...protein, critical for its fusion after binding to angiotensin-converting enzyme 2 (ACE2), as antiviral targets. We first validate cleavage at a putative furin substrate motif at SARS-CoV-2 spikes by expressing it in VeroE6 cells and find prominent syncytium formation. Cleavage and the syncytium are abolished by treatment with the furin inhibitors decanoyl-RVKR-chloromethylketone (CMK) and naphthofluorescein, but not by the transmembrane protease serine 2 (TMPRSS2) inhibitor camostat. CMK and naphthofluorescein show antiviral effects on SARS-CoV-2-infected cells by decreasing virus production and cytopathic effects. Further analysis reveals that, similar to camostat, CMK blocks virus entry, but it further suppresses cleavage of spikes and the syncytium. Naphthofluorescein acts primarily by suppressing viral RNA transcription. Therefore, furin inhibitors may be promising antiviral agents for prevention and treatment of SARS-CoV-2 infection.
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•The furin cleavage site in the SARS-CoV-2 spike protein mediates syncytium formation•The SARS-CoV-2 spike-mediated syncytium is suppressed by specific furin inhibitors•Furin inhibitors block SARS-CoV-2 virus entry and virus replication•Furin inhibitors are potential antiviral agents for SARS-CoV-2 infection and pathogenesis
Development of effective antiviral agents is an urgent unmet need for SARS-CoV-2 infection. Cheng et al. find that cleavage of the furin substrate site in the viral spike protein is critical for virus production and cytopathic effects. Two inhibitors targeting furin are potential antiviral agents to control SARS-CoV-2 infection and pathogenesis.
Softening is a common phenomenon of texture changes associated with plant cell walls, inducing a decrease in the quality of fruit. Inhibiting the softening is effective to extend the shelf life of ...fruit. Cold plasma (CP), as a novel nonthermal technology, has been applied to keep the freshness of the fruit. This review centers on applying cold plasma treatments to the inhibition of fruit softening. Different pathways for inhibiting fruit softening by CP treatments, including maintenance of fruit firmness, reduction in the activities of enzymes, inactivation of fungal pathogens and lowering of respiration rates, are discussed. The biochemistry of fruit softening and the fundamental of cold plasma are also presented. In general, among all postharvest technologies, cold plasma is a promising method with many advantages, showing great potential in maintaining the quality and inhibiting the softening of the fruit. Future work should focus on process optimization to achieve better results in maintaining fruit freshness, and commercial applications of cold plasma technology should also be explored.
A molecular nano‐floating gate (NFG) of pentacene‐based transistor memory devices is developed using conjugated polymer nanoparticles (CPN) as the discrete trapping sites embedded in an insulating ...polymer, poly (methacrylic acid) (PMAA). The nanoparticles of polyfluorene (PF) and poly(fluorene‐alt‐benzo2,1,3thiadiazole (PFBT) with average diameters of around 50–70 nm are used as charge‐trapping sites, while hydrophilic PMAA serves as a matrix and a tunneling layer. By inserting PF nanoparticles as the floating gate, the transistor memory device reveals a controllable threshold voltage shift, indicating effectively electron‐trapping by the PF CPN. The electron‐storage capability can be further improved using the PFBT‐based NFG since their lower unoccupied molecular orbital level is beneficial for stabilization of the trapped charges, leading a large memory window (35 V), retention time longer than 104 s with a high ON/OFF ratio of >104. In addition, the memory device performance using conjugated polymer nanoparticle NFG is much higher than that of the corresponding polymer blend thin films of PF/polystyrene. It suggests that the discrete polymer nanoparticles can be effectively covered by the tunneling layer, PMAA, to achieve the superior memory characteristics.
A molecular nano‐floating gate approach using conjugated polymer nanoparticles is developed for achieving high performance transistor memory devices. The transistor memory device using discrete polyfluorene and poly(fluorene‐alt‐benzo2,1,3thiadiazole nanoparticles as the floating gates can effectively trap the electrons and lead to a large memory window, long retention time, and a high ON/OFF ratio of >104.
Heavy metal ions are highly toxic and widely spread as environmental pollutants. New strategies are being developed to simultaneously detect and remove these toxic ions. Herein, we take the intrinsic ...advantage of covalent organic frameworks (COFs) and develop fluorescent COFs for sensing applications. As a proof-of-concept, a thioether-functionalized COF material, COF-LZU8, was “bottom-up” integrated with multifunctionality for the selective detection and facile removal of mercury(II): the π-conjugated framework as the signal transducer, the evenly and densely distributed thioether groups as the Hg2+ receptor, the regular pores facilitating the real-time detection and mass transfer, together with the robust COF structure for recycle use. The excellent sensing performance of COF-LZU8 was achieved in terms of high sensitivity, excellent selectivity, easy visibility, and real-time response. Meanwhile, the efficient removal of Hg2+ from water and the recycling of COF-LZU8 offers the possibility for practical applications. In addition, X-ray photoelectron spectroscopy and solid-state NMR investigations verified the strong and selective interaction between Hg2+ and the thioether groups of COF-LZU8. This research not only demonstrates the utilization of fluorescent COFs for both sensing and removal of metal ions but also highlights the facile construction of functionalized COFs for environmental applications.
Hemophilia A, a bleeding disorder resulting from F8 mutations, can only be cured by gene therapy. A promising strategy is CRISPR-Cas9-mediated precise insertion of F8 in hepatocytes at highly ...expressed gene loci, such as albumin (Alb). Unfortunately, the precise in vivo integration efficiency of a long insert is very low (~ 0.1%).
We report that the use of a double-cut donor leads to a 10- to 20-fold increase in liver editing efficiency, thereby completely reconstituting serum F8 activity in a mouse model of hemophilia A after hydrodynamic injection of Cas9-sgAlb and B domain-deleted (BDD) F8 donor plasmids. We find that the integration of a double-cut donor at the Alb locus in mouse liver is mainly through non-homologous end joining (NHEJ)-mediated knock-in. We then target BDDF8 to multiple sites on introns 11 and 13 and find that NHEJ-mediated insertion of BDDF8 restores hemostasis. Finally, using 3 AAV8 vectors to deliver genome editing components, including Cas9, sgRNA, and BDDF8 donor, we observe the same therapeutic effects. A follow-up of 100 mice over 1 year shows no adverse effects.
These findings lay the foundation for curing hemophilia A by NHEJ knock-in of BDDF8 at Alb introns after AAV-mediated delivery of editing components.
Polymers with an abundant amorphous domain should facilitate energy dissipation upon stretching, making near amorphous π-conjugated polymers have immense potential in realizing intrinsically ...stretchable field-effect transistor (FET) devices. In this study, high mobility preservation under the stretched state is attempted by replacing typical alkyl-monothienyl (T-R) on a benzo1,2-b:4,5-b’dithiophene-difluorobenzothiadiazole backbone with three other biaxially extended side-chains, including alkyl-dithienyl (2T-R), branching alkyl-trithienyl (3T-R), and alkyl-benzotrithienyl (B3T-R) groups. Despite showing near amorphous features, the semi-2D BDT-based polymers with bulkier biaxially extended side chains (PBDT-2T, PBDT-3T, and PBDT-B3T) still present comparable mobility to the reference semicrystalline polymer (PBDT-T). Although these four polymers yield comparable mobility, they show distinctly different mobility retention in the stretched state. From the study of their mobility-stretchability relationship, the interdigitating and/or entanglement of these biaxially extended conjugated side chains are shown to play a nontrivial role in the resultant mechanical robustness against the stretching force. Owing to the proper spatial mobility and geometry, the branched 3T-R side chain possesses a more intense interdigitating and/or entanglement capability than the linear 2T-R one and the fused B3T-R one, providing better mechanical strength under stretched states. Meanwhile, it maintains sufficient interchain connectivity for intermittent interchain hopping to compensate for the 1D charge transport along the backbone, ensuring good charge transport even in the stretched state. As a result, the printed PBDT-3T film is demonstrated to deliver a high mobility retention of 73% at a 60% strain exerted parallel to the charge-transporting direction and a very stable mobility retention of 88% after 1000 stretching-releasing cycles at a 60% strain, being one of the best stretchable near amorphous conjugated polymers reported thus far. Our result underlines the effectiveness of using biaxially extended conjugated side chains to realize high-performance stretchable polymers.
Robots can benefit education in many ways, and robotics holds great promise as a learning technology. However, the use of robots in education remains largely unknown to both researchers and ...educators. Many educators and developers have questions regarding the essential applications for robots used in education. Thus, this study aims to identify the essential applications of educational robots. To gain insight from the perspectives of researchers, experts and educators, we investigate the requirements of educational robots in all levels of education through 3 approaches: systematic literature review, expert interviews and instructor surveys. We review the extant literature that addresses the use of robots in education and adopted data from one of them, and conduct interviews with industry experts, scholars and instructors. We also conduct an online survey that invites instructors from six different levels of education to respond. We find the following to be the top five essential applications for educational robots: language education, robotics education, teaching assistance, social skill development and special education, and guided learning through feedback.
•An investigation that identifies the essential applications of educational robots.•The investigation consists of a systematic literature review, expert interviews and instructor surveys.•Robotics, language education and robot teaching assistant are the top three applications.•Robotics, language education and social and special education are the core applications.•Preschool and primary school group have greater potential to implement educational robots in near future.
Abstract
Investigations of CRISPR gene knockout editing profiles have contributed to enhanced precision of editing outcomes. However, for homology-directed repair (HDR) in particular, the editing ...dynamics and patterns in clinically relevant cells, such as human iPSCs and primary T cells, are poorly understood. Here, we explore the editing dynamics and DNA repair profiles after the delivery of Cas9-guide RNA ribonucleoprotein (RNP) with or without the adeno-associated virus serotype 6 (AAV6) as HDR donors in four cell types. We show that editing profiles have distinct differences among cell lines. We also reveal the kinetics of HDR mediated by the AAV6 donor template. Quantification of T50 (time to reach half of the maximum editing frequency) indicates that short indels (especially +A/T) occur faster than longer (>2 bp) deletions, while the kinetics of HDR falls between NHEJ (non-homologous end-joining) and MMEJ (microhomology-mediated end-joining). As such, AAV6-mediated HDR effectively outcompetes the longer MMEJ-mediated deletions but not NHEJ-mediated indels. Notably, a combination of small molecular compounds M3814 and Trichostatin A (TSA), which potently inhibits predominant NHEJ repairs, leads to a 3-fold increase in HDR efficiency.
In brain-computer interface (BCI) work, how correctly identifying various features and their corresponding actions from complex Electroencephalography (EEG) signals is a challenging technology. ...However, most current methods do not consider EEG feature information in spatial, temporal and spectral domains, and the structure of these models cannot effectively extract discriminative features, resulting in limited classification performance. To address this issue, we propose a novel text motor-imagery EEG discrimination method, namely wavelet-based temporal-spectral-attention correlation coefficient (WTS-CC), to simultaneously consider the features and their weighting in spatial, EEG-channel, temporal and spectral domains in this study. The initial Temporal Feature Extraction (iTFE) module extracts the initial important temporal features of MI EEG signals. The Deep EEG-Channel-attention (DEC) module is then proposed to automatically adjust the weight of each EEG channel according to its importance, thereby effectively enhancing more important EEG channels and suppressing less important EEG channels. Next, the Wavelet-based Temporal-Spectral-attention (WTS) module is proposed to obtain more significant discriminative features between different MI tasks by weighting features on two-dimensional time-frequency maps. Finally, a simple discrimination module is used for MI EEG discrimination. The experimental results indicate that the proposed text WTS-CC method can achieve promising discrimination performance that outperforms the state-of-the-art methods in terms of classification accuracy, Kappa coefficient, F1 score, and AUC on three public datasets.
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