Nanoconstruction on Living Cell Surfaces with Cucurbit[7]uril-Based Supramolecular Polymer Chemistry: Toward Cell-Based Delivery of Bio-Orthogonal Catalytic Systems.

Employing living cells as carriers to transport transition metal-based catalysts for target-specific bio-orthogonal catalysis represents a cutting-edge approach in advancing precision biomedical applications. One of the initial hurdles in this endeavor involves effectively attaching the catalysts to the carrier cells while preserving the cells' innate ability to interact with biological systems and maintaining the unaltered catalytic activity. In this study, we have developed an innovative layer-by-layer method that leverages a noncovalent interaction between cucurbit[7]uril and adamantane as the primary driving force for crafting polymeric nanostructures on the surfaces of these carrier cells. The strong binding affinity between the host-guest pair ensures the creation of a durable polymer coating on the cell surfaces. Meanwhile, the layer-by-layer process offers high adaptability, facilitating the efficient loading of bio-orthogonal catalysts onto cell surfaces. Importantly, the polymeric coating shows no discernible impact on the cells' physiological characteristics, including their tropism, migration, and differentiation, while preserving the effectiveness of the bio-orthogonal catalysts.

Exploring the supramolecular chemistry of cyclopropeniums: halogen-bonding-induced electrostatic assembly of polymers.

Exploring new noncovalent synthons for supramolecular assembly is essential for material innovation. Accordingly, we herein report a unique type of cyclopropenium-based supramolecular motif and demonstrate its applications to polymer self-assembly. Because of the "ion pair strain" effect, trisaminocyclopropenium iodides complex strongly with fluoroiodobenzene derivatives, forming stable adducts. Crystal structure analysis reveals that halogen-bonding between the iodide anion and the iodo substituent of the fluoroiodobenzene is the driving force for the formation of these electrostatically complexed adducts. Such halogen-bonding-induced electrostatic interactions were further successfully applied to drive the assembly of polymers in solution, on surfaces, and in bulk, demonstrating their potential for constructing supramolecular polymeric materials.

Inspection and maintenance optimization for heterogeneity units in redundant structure with Non-dominated Sorting Genetic Algorithm III.

Redundant structure has been widely deployed to improve system reliability, as when one unit fails, the system can continue to function by using another one. Most existing studies rely on the similar assumption that the heterogeneous units are subject to periodic inspections and identical in terms of their aging situations and the numbers of resisted shocks. In practice, it is often adequate to trigger a unit individually in the event of a single shock, which intensifies the degradation of that unit, accordingly, requiring a sooner inspection to ensure its safety. In this study, the stochastic dependency among units is addressed firstly by introducing a novel activation sequence. Secondly, an adaptive system-level inspection policy is proposed by prioritizing the unit with a worse state. Finally, we take advantage of Monte Carlo methods to simulate the whole process and estimate two objectives, referring to the average system unavailability and maintenance cost, in a designed service time. It is found that the two objectives are contradictory through numerical examples. The Non-dominated Sorting Genetic Algorithm III (NSGA-III) algorithm, therefore, has been employed to find the optimal solutions in system unavailability and cost, which provide clues for practitioners in decision-making.

Promoting halogen-bonding catalyzed living radical polymerization through ion-pair strain.

Discovering efficient catalysts is highly desired in expanding the application of halogen-bonding catalysis. We herein report our findings on applying triaminocyclopropenium (TAC) iodides as highly potent catalysts for halogen-bonding catalyzed living radical polymerization. Promoted by the unique effect of ion-pair strain between the TAC cation and the iodide anion, the TAC iodides showed high catalytic efficiency in the halogen-bonding catalysis toward radical generation, and surpassed the previously reported organic iodide catalysts. With the TAC iodide as catalyst, radical polymerization with a living feature was successfully realized, which shows general applicability with a variety of monomers and produced block copolymers. In addition, the TAC-iodides also showed promising feasibility in catalyzing the radical depolymerization of iodo-terminated polymethacrylates. Noteworthily, the catalytic capacity of the TAC iodides is demonstrated to be closely related to the electronic properties of the TAC cation, which offers a molecular platform for further catalyst screening and optimization.

Engineering living cells with cucurbit[7]uril-based supramolecular polymer chemistry: from cell surface engineering to manipulation of subcellular organelles.

Supramolecular polymer chemistry, which closely integrates noncovalent interactions with polymeric structures, is a promising toolbox for living cell engineering. Here, we report our recent progress in exploring the applications of cucurbit[7]uril (CB[7])-based supramolecular polymer chemistry for engineering living cells. First, a modular polymer-analogous approach was established to prepare multifunctional polymers that contain CB[7]-based supramolecular recognition motifs. The supramolecular polymeric systems were successfully applied to cell surface engineering and subcellular organelle manipulation. By anchoring polymers on the cell membranes, cell-cell interactions were established by CB[7]-based host-guest recognition, which further facilitated heterogeneous cell fusion. In addition to cell surface engineering, placing the multifunctional polymers on specific subcellular organelles, including the mitochondria and endoplasmic reticulum, has led to enhanced physical contact between subcellular organelles. It is highly anticipated that the CB[7]-based supramolecular polymer chemistry will provide a new strategy for living cell engineering to advance the development of cell-based therapeutic materials.

Performance assessment of K-out-of-N safety instrumented systems subject to cascading failures.

Safety instrumented systems often employ redundancy to enhance the ability to detect and respond to hazardous events. The use of redundancy increases the fault tolerance to single failure but remains vulnerable in case of dependent failures, including common cause failures and cascading failures. Reliability analysis of safety instrumented systems therefore involves the impact of dependent failures. The used approaches have primarily focused on common cause failures. In this paper, it is argued the need to consider the efforts of cascading failures that are caused by functional dependencies, hazardous events, and shared resources. A recursive aggregation-based approach is proposed for performance analyzing of K-out-of-N safety instrumented systems with consideration of cascading failures. General approximation formulas are developed for estimating the average probability of failures on demand of different configurations of safety instrumented systems. These formulas are compared with those for common cause failures. Then a case of fire water pump is studied to illustrate the effects of cascading failures on safety instrumented systems.

Fusion of Infrared and Visible Images Using Fast Global Smoothing Decomposition and Target-Enhanced Parallel Gaussian Fuzzy Logic.

As a powerful technique to merge complementary information of original images, infrared (IR) and visible image fusion approaches are widely used in surveillance, target detecting, tracking, and biological recognition, etc. In this paper, an efficient IR and visible image fusion method is proposed to simultaneously enhance the significant targets/regions in all source images and preserve rich background details in visible images. The multi-scale representation based on the fast global smoother is firstly used to decompose source images into the base and detail layers, aiming to extract the salient structure information and suppress the halos around the edges. Then, a target-enhanced parallel Gaussian fuzzy logic-based fusion rule is proposed to merge the base layers, which can avoid the brightness loss and highlight significant targets/regions. In addition, the visual saliency map-based fusion rule is designed to merge the detail layers with the purpose of obtaining rich details. Finally, the fused image is reconstructed. Extensive experiments are conducted on 21 image pairs and a Nato-camp sequence (32 image pairs) to verify the effectiveness and superiority of the proposed method. Compared with several state-of-the-art methods, experimental results demonstrate that the proposed method can achieve more competitive or superior performances according to both the visual results and objective evaluation.

Evaluation of IoT-Enabled Monitoring and Electronic Nose Spoilage Detection for Salmon Freshness During Cold Storage.

Salmon is a highly perishable food due to temperature, pH, odor, and texture changes during cold storage. Intelligent monitoring and spoilage rapid detection are effective approaches to improve freshness. The aim of this work was an evaluation of IoT-enabled monitoring system (IoTMS) and electronic nose spoilage detection for quality parameters changes and freshness under cold storage conditions. The salmon samples were analyzed and divided into three groups in an incubator set at 0 °C, 4 °C, and 6 °C. The quality parameters, i.e., texture, color, sensory, and pH changes, were measured and evaluated at different temperatures after 0, 3, 6, 9, 12, and 14 days of cold storage. The principal component analysis (PCA) algorithm can be used to cluster electronic nose information. Furthermore, a Convolutional Neural Networks and Support Vector Machine (CNN-SVM) based algorithm is used to cluster the freshness level of salmon samples stored in a specific storage condition. In the tested samples, the results show that the training dataset of freshness is about 95.6%, and the accuracy rate of the test dataset is 93.8%. For the training dataset of corruption, the accuracy rate is about 91.4%, and the accuracy rate of the test dataset is 90.5%. The overall accuracy rate is more than 90%. This work could help to reduce quality loss during salmon cold storage.

Catalytically Active Single-Chain Polymeric Nanoparticles: Exploring Their Functions in Complex Biological Media.

Dynamic single-chain polymeric nanoparticles (SCPNs) are intriguing, bioinspired architectures that result from the collapse or folding of an individual polymer chain into a nanometer-sized particle. Here we present a detailed biophysical study on the behavior of dynamic SCPNs in living cells and an evaluation of their catalytic functionality in such a complex medium. We first developed a number of delivery strategies that allowed the selective localization of SCPNs in different cellular compartments. Live/dead tests showed that the SCPNs were not toxic to cells while spectral imaging revealed that SCPNs provide a structural shielding and reduced the influence from the outer biological media. The ability of SCPNs to act as catalysts in biological media was first assessed by investigating their potential for reactive oxygen species generation. With porphyrins covalently attached to the SCPNs, singlet oxygen was generated upon irradiation with light, inducing spatially controlled cell death. In addition, Cu(I)- and Pd(II)-based SCPNs were prepared and these catalysts were screened in vitro and studied in cellular environments for the carbamate cleavage reaction of rhodamine-based substrates. This is a model reaction for the uncaging of bioactive compounds such as cytotoxic drugs for catalysis-based cancer therapy. We observed that the rate of the deprotection depends on both the organometallic catalysts and the nature of the protective group. The rate reduces from in vitro to the biological environment, indicating a strong influence of biomolecules on catalyst performance. The Cu(I)-based SCPNs in combination with the dimethylpropargyloxycarbonyl protective group showed the best performances both in vitro and in biological environment, making this group promising in biomedical applications.

Asiaticoside Attenuates Cell Growth Inhibition and Apoptosis Induced by Aß1-42 via Inhibiting the TLR4/NF-κB Signaling Pathway in Human Brain Microvascular Endothelial Cells.

Alzheimer's disease (AD) is a very common progressive neurodegenerative disorder with the highest incidence in the world. Dysfunction of the blood-brain barrier (BBB) may be responsible for the pathogenesis and pathology of AD for abnormally transporting amyloid-ß (Aß, the main component of the senile plaques) from the sera into the central nervous system. Aß peptides induce apoptosis in human brain microvascular endothelial cells (hBMECs), the main component of BBB. Apoptosis in neuronal cells plays a critical role in the pathogenesis of AD. Asiaticoside, a natural glycoside extracted from Centella asiatica (L.) Urban, has an anti-apoptotic effect on hBMECs but the molecule mechanism remains unclear. Therefore, we investigate the protective effect of asiaticoside on Aß1-42-induced cytotoxicity and apoptosis as well as associated mechanism in hBMECs with commonly used in vitro methods for clinical development of asiaticoside as a novel anti-AD agent. In the present study, we investigated the effects of asiaticoside on cytotoxicity by Cell Counting Kit-8 assay, mitochondrial membrane potential by JC-1 fluorescence analysis, anti-apoptosis by Hoechst 33258 staining and Annexin V-FITC (fluorescein isothiocyanate) and propidium iodide (PI) analyses, the expressions of TNF-α and IL-6 by enzyme-linked immunosorbent assay (ELISA) and TLR4, MyD88, TRAF6, p-NF-κB p65, and total NF-κB p65 by Western blotting, and nuclear translocation of NF-κB p65 by immunofluorescence analysis in hBMECs. The results showed that pretreatment of asiaticoside (25, 50, and 100 µM) for 12 h significantly attenuated cell growth inhibition and apoptosis, and restored declined mitochondrial membrane potential induced by Aß1-42 (50 µM) in hBMECs. Asiaticoside also significantly downregulated the elevated expressions of TNF-α, IL-6, TLR4, MyD88, TRAF6, and p-NF-κB p65, as well as inhibited NF-κB p65 translocation from cytoplasm to nucleus induced by Aß1-42 in hBMECs in a concentration-dependent manner. The possible underlying molecular mechanism of asiaticoside may be through inhibiting the TLR4/NF-κB signaling pathway. Therefore, asiaticoside may be developed as a novel agent for the prevention and/or treatment of AD clinically.

Improving the Folding of Supramolecular Copolymers by Controlling the Assembly Pathway Complexity.

A family of amphiphilic, heterograft copolymers containing hydrophilic, hydrophobic, and supramolecular units based on Jeffamine M-1000, dodecylamine, and benzene-1,3,5-tricarboxamide (BTA) motifs, respectively, was prepared via a postfunctionalization approach. The folding of the copolymers in water into nanometer-sized particles was analyzed by a combination of dynamic and static light scattering, circular dichroism spectroscopy, and small-angle neutron scattering. The sample preparation protocol was crucial for obtaining reproducible and consistent results, showing that only full control over the structure and pathway complexity will afford the desired folded structure, a phenomenon similar to protein folding. The results revealed that relatively small changes in the polymer's graft composition strongly affected the intra- versus intermolecular assembly processes. Depending on the amount of the hydrophobic grafts based on either dodecyl or BTA groups, pronounced behavioral differences were observed for copolymers that comprise similar degrees of hydrophobic content. A high number of BTA grafts (>10%) resulted in the formation of multichain aggregates comprising around six polymer chains. In contrast, for copolymers comprising up to 10% BTA grafts the folding results in nanoparticles that adopt open, sparse conformations and comprise one to two polymer chains. Interestingly, predominantly single-chain polymeric nanoparticles were formed when the copolymer comprised only Jeffamine or Jeffamine and dodecyl grafts. In addition, replacing part of the BTA grafts by hydrophobic dodecyl grafts while keeping the hydrophobic content constant promoted single-chain folding and resulted in the formation of a compact, globular nanoparticle with a more structured interior. Thus, the intra- and intermolecular self-assembly pathways can be directed by carefully tuning the polymer's hydrophilic-hydrophobic balance in combination with the number of supramolecular grafts.

Activation of Nrf2 Signaling Augments Vesicular Stomatitis Virus Oncolysis via Autophagy-Driven Suppression of Antiviral Immunity.

Oncolytic viruses (OVs) offer a promising therapeutic approach to treat multiple types of cancer. In this study, we show that the manipulation of the antioxidant network via transcription factor Nrf2 augments vesicular stomatitis virus Δ51 (VSVΔ51) replication and sensitizes cancer cells to viral oncolysis. Activation of Nrf2 signaling by the antioxidant compound sulforaphane (SFN) leads to enhanced VSVΔ51 spread in OV-resistant cancer cells and improves the therapeutic outcome in different murine syngeneic and xenograft tumor models. Chemoresistant A549 lung cancer cells that display constitutive dominant hyperactivation of Nrf2 signaling are particularly vulnerable to VSVΔ51 oncolysis. Mechanistically, enhanced Nrf2 signaling stimulated viral replication in cancer cells and disrupted the type I IFN response via increased autophagy. This study reveals a previously unappreciated role for Nrf2 in the regulation of autophagy and the innate antiviral response that complements the therapeutic potential of VSV-directed oncolysis against multiple types of OV-resistant or chemoresistant cancer.

RIGulation of STING expression: at the crossroads of viral RNA and DNA sensing pathways.

The innate immune sensing of pathogens is important for host to mount defensive responses. STING has emerged in recent years as a critical signaling adaptor in the immune response to cytosolic DNA and RNA derived from pathogens. Liu et al. (2016) demonstrate that the RIG-I-dependent RNA sensing signaling induces STING expression via a TNF-α and IFN-α synergy. The up-regulation of STING is vital for 5'pppRNA restriction of HSV, a DNA virus that infects humans and causes herpes, in vitro and in vivo. This study provides new insights into the cross talk between DNA and RNA pathogen-sensing systems via the control of STING.

ArfGAP Domain-Containing Protein 2 (ADAP2) Integrates Upstream and Downstream Modules of RIG-I Signaling and Facilitates Type I Interferon Production.

Transcription of type I interferon genes during RNA virus infection requires signal communication between several pattern recognition receptor (PRR)-adaptor complexes located at distinct subcellular membranous compartments and a central cytoplasmic TBK1-interferon regulatory factor 3 (IRF3) kinase-transcription factor module. However, how the cell integrates signal transduction through spatially distinct modules of antiviral signaling pathways is less defined. RIG-I is a major cytosolic PRR involved in the control of several RNA viruses. Here we identify ArfGAP domain-containing protein 2 (ADAP2) as a key novel scaffolding protein that integrates different modules of the RIG-I pathway, located at distinct subcellular locations, and mediates cellular antiviral type I interferon production. ADAP2 served to bridge the mitochondrial membrane-bound upstream RIG-I adaptor MAVS and the downstream cytosolic complex of NEMO (regulatory subunit of TBK1), TBK1, and IRF3, leading to IRF3 phosphorylation. Furthermore, independently, ADAP2 also functioned as a major orchestrator of the interaction of TBK1 with NEMO and IRF3. Mutational and in vitro cell-free reconstituted RIG-I signaling assay-based analyses identified that the ArfGAP domain of ADAP2 mediates the interferon response. TRAF3 acted as a trigger for ADAP2 to recruit RIG-I pathway component proteins into a single macromolecular complex. This study provides important novel insights into the assembly and integration of different modules of antiviral signaling cascades.

RIG-I-Mediated STING Upregulation Restricts Herpes Simplex Virus 1 Infection.

UNLABELLED: STING has emerged in recent years as a key player in orchestrating innate immune responses to cytosolic DNA and RNA derived from pathogens. However, the regulation of STING still remains poorly defined. In the present study, we investigated the mechanism of the regulation of STING expression in relation to the RIG-I pathway. Our data show that signaling through RIG-I induces STING expression at both the transcriptional and protein levels in various cell types. STING induction by the RIG-I agonist 5'triphosphorylated RNA (5'pppRNA) was recognized to be a delayed event resulting from an autocrine/paracrine mechanism. Indeed, cotreatment with tumor necrosis factor alpha and type I/II interferon was found to have a synergistic effect on the regulation of STING expression and could be potently decreased by impairing NF-κB and/or STAT1/2 signaling. STING induction significantly contributed to sustainment of the immune signaling cascade following 5'pppRNA treatment. Physiologically, this cross talk between the RNA- and DNA-sensing pathways allowed 5'pppRNA to efficiently block infection by herpes simplex virus 1 (HSV-1) both in vitro and in vivo in a STING-dependent fashion. These observations demonstrate that STING induction by RIG-I signaling through the NF-κB and STAT1/2 cascades is essential for RIG-I agonist-mediated HSV-1 restriction. IMPORTANCE: The innate immune system represents the first line of defense against invading pathogens. The dysregulation of this system can result in failure to combat pathogens, inflammation, and autoimmune diseases. Thus, precise regulation at each level of the innate immune system is crucial. Recently, a number of studies have established STING to be a central molecule in the innate immune response to cytosolic DNA and RNA derived from pathogens. Here, we describe the regulation of STING via RIG-I-mediated innate immune sensing. We found that STING is synergistically induced via proinflammatory and antiviral cytokine cascades. In addition, we show that in vivo protection against herpes simplex virus 1 (HSV-1) by a RIG-I agonist required STING. Our study provides new insights into the cross talk between DNA and RNA pathogen-sensing systems via the control of STING.

Host and Viral Modulation of RIG-I-Mediated Antiviral Immunity.

Innate immunity is the first line of defense against invading pathogens. Rapid and efficient detection of pathogen-associated molecular patterns via pattern-recognition receptors is essential for the host to mount defensive and protective responses. Retinoic acid-inducible gene-I (RIG-I) is critical in triggering antiviral and inflammatory responses for the control of viral replication in response to cytoplasmic virus-specific RNA structures. Upon viral RNA recognition, RIG-I recruits the mitochondrial adaptor protein mitochondrial antiviral signaling protein, which leads to a signaling cascade that coordinates the induction of type I interferons (IFNs), as well as a large variety of antiviral interferon-stimulated genes. The RIG-I activation is tightly regulated via various posttranslational modifications for the prevention of aberrant innate immune signaling. By contrast, viruses have evolved mechanisms of evasion, such as sequestrating viral structures from RIG-I detections and targeting receptor or signaling molecules for degradation. These virus-host interactions have broadened our understanding of viral pathogenesis and provided insights into the function of the RIG-I pathway. In this review, we summarize the recent advances regarding RIG-I pathogen recognition and signaling transduction, cell-intrinsic control of RIG-I activation, and the viral antagonism of RIG-I signaling.

Modular Synthetic Platform for the Construction of Functional Single-Chain Polymeric Nanoparticles: From Aqueous Catalysis to Photosensitization.

Single-chain polymeric nanoparticles (SCPNs) are intriguing systems for multiple applications. In order to arrive at a controlled, but random, positioning of the different side groups to the polymer backbone, alternative synthetic routes have to be developed. Here, a general postpolymerization modification strategy of poly(pentafluorophenyl acrylate) (pPFPA) is presented as a versatile method to rapidly access functional SCPNs. We first show that the sequential addition of a benzene-1,3,5-tricarboxamide-based amine, acting as the supramolecular recognition motif, and water-soluble polyetheramine (Jeffamine) to pPFPA affords random copolymers that fold in water into SCPNs. The scope of the modular platform is illustrated by preparing two types of functional SCPNs. First, we prepared SCPNs designed for bio-orthogonal catalysis by attaching pendant mono(benzimidazoylmethyl)-bis(pyridylmethyl) (Bimpy), phenanthroline (Phen), or 2,2'-bipyridine (BiPy), ligands capable of binding either Cu(I) or Pd(II). The Bimpy- and Phen-containing SCPNs ligated to Cu(I) significantly accelerate azide-alkyne cycloaddition reactions while Bipy-containing SCPNs ligated to Pd(II) efficiently catalyze depropargylation reactions. In all cases, reactions proceeded efficiently in phosphate buffer at a physiological pH and at low substrate concentrations. Next, the potential of SCPNs for photodynamic therapy was evaluated. Introducing porphyrins in SCPNs leads to novel photosensitizers that can produce singlet oxygen ((1)O2) upon photoirradiation. Additionally, by attaching both porphyrins and prodrug models, attached via (1)O2-cleavable amino-acrylate linker, to the SCPNs, we show that irradiation of the SCPNs results in a cascade reaction of (1)O2 generation followed by cleavage of the amino-acrylate linkers, releasing the drug model. The modular synthesis strategy reported here provides rapid and controlled access to SCPNs with tunable amounts of active units that fulfill different functions.

Supramolecular polymerization promoted and controlled through self-sorting.

A new method in which supramolecular polymerization is promoted and controlled through self-sorting is reported. The bifunctional monomer containing p-phenylene and naphthalene moieties was prepared. Supramolecular polymerization is promoted by selective recognition between the p-phenylene group and cucurbit[7]uril (CB[7]), and 2:1 complexation of the naphthalene groups with cucurbit[8]uril (CB[8]). The process can be controlled by tuning the CB[7] content. This development will enrich the field of supramolecular polymers with important advances towards the realization of molecular-weight and structural control.

Rational adjustment of multicolor emissions by cucurbiturils-based host-guest chemistry and photochemistry.

The host-guest chemistry of cucurbiturils and the photochemistry of azastilbene derivatives are combined for the rationally adjusting multicolor emissions through forming different host-guest complexes and their corresponding photochemical products. Cucurbit[8]uril (CB[8]) can bind with azastilbene derivatives to form supramolecular polymers emitting orange light. The supramolecular polymers further facilitate the [2 + 2] cycloaddition of C═C bonds in azastilbenes by UV irradiation, emitting blue light. Different from CB[8], cucurbit[7]uril (CB[7]) encapsulates azastilbene derivatives to form a dumbbell-shaped host-guest complex, emitting dark-purple light. This dumbbell-shaped host-guest complex undergoes cis-isomerization after UV irradiation, thus emitting green light. Therefore, this strategy is promising for fabricating advanced stimuli-responsive fluorescent materials.