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Papain-like protease (PLpro) is an attractive anti-coronavirus target. The development of PLpro inhibitors, however, is hampered by the limitations of the existing PLpro assay and the scarcity of validated active compounds. We developed a novel in-cell PLpro assay based on BRET and used it to evaluate and discover SARS-CoV-2 PLpro inhibitors. The developed assay demonstrated remarkable sensitivity for detecting the reduction of intracellular PLpro activity while presenting high reliability and performance for inhibitor evaluation and high-throughput screening. Using this assay, three protease inhibitors were identified as novel PLpro inhibitors that are structurally disparate from those previously known. Subsequent enzymatic assays and ligand-protein interaction analysis based on molecular docking revealed that ceritinib directly inhibited PLpro, showing high geometric complementarity with the substrate-binding pocket in PLpro, whereas CA-074 methyl ester underwent intracellular hydrolysis, exposing a free carboxyhydroxyl group essential for hydrogen bonding with G266 in the BL2 groove, resulting in PLpro inhibition. [ABSTRACT FROM AUTHOR]

The potential of chimeric antigen receptor (CAR)-based immunotherapy as a promising therapeutic approach is often hindered by the presence of highly immunosuppressive tumor microenvironments (TME). Combination therapies with either co-administration or built-in expression of additional TME-modulating therapeutic molecules to potentiate the functions of CAR-T cells can cause systemic toxicities due to the lack of control over the delivery of biologics. Here, we present a proof-of-concept engineered platform in human Jurkat T cells that combines CAR with a therapeutic gene circuit capable of sensing β-galactosidase (a reported cancer-associated signal) and subsequently activate the production of customized therapeutic gene products. We have demonstrated the integration of the chemically induced proximity (CIP) and associated signal sensing technologies with CAR in this study. A β-galactosidase-activatable prodrug was designed by conjugating a galactose moiety with a CIP inducer abscisic acid (ABA). We showed that Jurkat T cells engineered with CAR and the ABA-inducible genetic circuits can respond to recombinant β-galactosidase to drive the production and secretion of various immunotherapeutics including an anti-cancer agent, an immunomodulatory cytokine, and immune checkpoint inhibitors. Our design is highly modular and could be adapted to sense different cancer-related signals to locally produce antitumor therapeutics that can potentially boost CAR-T efficacy and persistence. [ABSTRACT FROM AUTHOR]

Change detection is a crucial task in remote sensing that finds broad application in land resource planning, forest resource monitoring, natural disaster monitoring, and evaluation. In this paper, we propose a change detection model for cross-domain recognition, which we call CrossCDNet. Our model significantly improves the modeling ability of the change detection on one dataset and demonstrates good generalization on another dataset without any additional operations. To achieve this, we employ a Siamese neural network for change detection and design an IBNM (Instance Normalization and Batch Normalization Module) that utilizes instance normalization and batch normalization in order to serve as the encoder backbone in the Siamese neural network. The IBNM extracts feature maps for each layer, and the Siamese neural network fuses the feature maps of the two branches using a unique operation. Finally, a simple MLP decoder is used for end-to-end change detection. We train our model on the LEVIR-CD dataset and achieve competitive performance on the test set. In cross-domain dataset testing, CrossCDNet outperforms all the other compared models. Specifically, our model achieves an F1-score of 91.69% on the LEVIR-CD dataset and an F1-score of 77.09% on the WHU-CD dataset, where the training set was LEVIR-CD. [ABSTRACT FROM AUTHOR]

Purpose - The purpose of this study is to investigate Western and Central European consumers and web retailers with respect to the importance and accessibility of delivery information online prior to purchase. Design/methodology/approach - To investigate the importance and accessibility of delivery information online, a survey was conducted of 715 internet consumers from the UK, Hungary and the Czech Republic to explore how important they rate online delivery information. In addition to this, retailing web sites from leading retailers in the UK, Hungary and the Czech Republic were content analysed in order to establish the extent to which they provided online delivery information. Findings - The research identifies that consumers rate delivery pricing guides, delivery guarantees and delivery schedules as the most important delivery information they expect online prior to purchase. However, content analysis of retailer web sites reveals that many retailers do not adequately provide information about how they ‘guarantee product delivery’. Practical implications - It is recommended here that prior to purchase online retailers should make detailed delivery information more accessible to consumers (e.g. pricing guides, timing schedules, etc.) and should consider using delivery service guarantees to assure consumers of delivery service standards and retailer responsibilities. Originality/value - This paper makes a decisive contribution to e-shopping behaviour and online retailing by providing insight into why visits to retailing web sites may not be followed up by purchase. This insight results from an examination of an often neglected area of the online buying process, namely ‘order delivery and fulfilment’. By examining the expectations of e-consumers across Europe it investigates the role access to delivery information can have in managing customer delivery service expectations and in building trust in online retailers. [ABSTRACT FROM AUTHOR]

Mutations in colony stimulating factor 1 receptor (CSF1R) result in CSF1R‐related disorder (CSF1R‐RD). Our previous study demonstrated a proteolytic generation of a soluble CSF1R (sCSF1R) that could potentially serve as a diagnostic biomarker of CSF1R‐RD. Herein, we observed that sCSF1R is released into peripheral serum as a highly glycosylated monomer in Csf1r+/− mice that mimic the clinical symptoms of CSF1R‐RD patients. Notably, we found that serum sCSF1R could distinguish CSF1R‐RD cohorts from controls with high accuracy as evaluated by receiver operating characteristic (ROC) curves. This study demonstrates that reduced sCSF1R in serum may serve as a diagnostic biomarker for CSF1R‐RD. ANN NEUROL 2024 [ABSTRACT FROM AUTHOR]

Ischemic stroke (IS) is a cerebrovascular sickness, and cerebral ischemia-reperfusion (I/R) damage often occurs, but there is still a lack of drugs that can significantly alleviate it. Curcumin (Cur) exerts pharmacological effects such as antioxidative stress, anti-inflammation, and the promotion of apoptosis through regulating various pathways, but its efficacy and specific mechanism of action in IS have not been fully clarified. The purpose of this paper is to study the influence of Cur on IS. Brain microvascular endothelial cells (BMECs) were used to create an oxygen-glucose deprivation/reoxygenation (OGD/R) model to simulate I/R damage. The cell viability was assessed using an MTT assay. The LDH level and ROS positive rate were measured using commercial kits. The cell invasion was examined using a transwell assay. The apoptosis was assessed by flow cytometry. The contents of GMFB, Bax, and Bcl2 were measured using western blot. We confirmed that in the OGD/R-induced IS cell model, the abundance of GMFB was enhanced in the OGD/R group versus the control group. GMFB overexpression promoted OGD/R-induced cell viability diminution, increased LDH and ROS levels, lessened cell invasion ability, enhanced cell apoptosis, enhanced Bax levels, and decreased Bcl2 levels. Silencing GMFB ameliorated OGD/R-induced cell damage. Cur ameliorated OGD/R-induced cell damage. Cur curbed OGD/R-induced cell damage by downregulating GMFB expression. In conclusion, Cur cured ischemic stroke-induced cell damage by downregulating GMFB expression.

Naphthoquine is a promising candidate for antimalarial combination therapy. Its combination with artemisinin has demonstrated excellent efficacy in clinical trials conducted across various malaria-endemic areas. A co-formulated combination of naphthoquine and azithromycin has also shown high clinical efficacy for malaria prophylaxis in Southeast Asia. Developing new combination therapies using naphthoquine will provide additional arsenal responses to the growing threat of artemisinin resistance. Furthermore, due to its long half-life, the possible interaction of naphthoquine with other drugs also needs attention. However, studies on its pharmacodynamic interactions with other drugs are still limited. In this study, the in vitro interactions of naphthoquine with ivermectin, atovaquone, curcumin, and ketotifen were evaluated in the asexual stage of Plasmodium falciparum 3D7. By using the combination index analysis and the SYBR Green I-based fluorescence assay, different interaction patterns of selected drugs with naphthoquine were revealed. Curcumin showed a slight but significant synergistic interaction with naphthoquine at lower effect levels, and no antagonism was observed across the full range of effect levels for all tested ratios. Atovaquone showed a potency decline when combined with naphthoquine. For ivermectin, a significant antagonism with naphthoquine was observed at a broad range of effect levels below 75% inhibition, although no significant interaction was observed at higher effect levels. Ketotifen interacted with naphthoquine similar to ivermectin, but significant antagonism was observed for only one tested ratio. These findings should be helpful to the development of new naphthoquine-based combination therapy and the clinically reasonable application of naphthoquine-containing therapies., Importance: Pharmacodynamic interaction between antimalarials is not only crucial for the development of new antimalarial combination therapies but also important for the appropriate clinical use of antimalarials. The significant synergism between curcumin and naphthoquine observed in this study suggests the potential value for further development of new antimalarial combination therapy. The finding of a decline in atovaquone potency in the presence of naphthoquine alerts to a possible risk of treatment or prophylaxis failure for atovaquone-proguanil following naphthoquine-containing therapies. The observation of antagonism between naphthoquine and ivermectin raised a need for concern about the applicability of naphthoquine-containing therapy in malaria-endemic areas with ivermectin mass drug administration deployed. Considering the role of atovaquone-proguanil as a major alternative when first-line artemisinin-based combination therapy is ineffective and the wide implementation of ivermectin mass drug administration in malaria-endemic countries, the above findings will be important for the appropriate clinical application of antimalarials involving naphthoquine-containing therapies., Competing Interests: The authors declare no conflict of interest.

Two cyclic carbazolenevinylene dimers 1 and 2 were synthesized by McMurry coupling reactions. A linear compound 3 was also prepared for comparison. Compounds 1-3 were fully characterized by means of NMR spectroscopy, HRMS, elemental analysis, and UV/Vis absorption spectroscopy. Quantum chemical simulations showed that the cyclic compounds possessed smaller HOMO-LUMO gaps and more extended conjugation. The UV/Vis absorption spectra of the cyclic compounds showed blueshifts compared with that of the linear compound 3. Time-dependent DFT (TD-DFT) analysis revealed that this was due to the different selection rules for molecules with cyclic and linear architectures. The cyclic conformation also significantly affected the molecular ordering in the solid state. The X-ray crystal structure of 1 showed partial pi-pi overlapping between the adjacent molecules. Thin films of 1-3 were fabricated by the vacuum-deposition method on Si/SiO(2) substrates. Multicrystalline thin films were obtained from compounds 1 and 2, but only amorphous thin films could be obtained for the linear compound 3. Another important difference between the cyclic and linear compounds was the reduced reorganization energy for the cyclic compounds. These two facts have resulted in improved field-effect transistor (FET) mobilities for the cyclic compounds compared with the linear compound. In addition, as the substrate temperature has a significant influence on the morphology and the degree of crystallinity of the thin films deposited, the device performance could be optimized by varying the substrate temperature. The FET devices based on 2 gave the highest mobility of 0.013 cm(2) V(-1) s(-1). The results showed that carbazole derivatives with cyclic structures might make better FETs.