Integration of covalent organic frameworks and molecularly imprinted polymers for selective extraction of flavonoid naringenin from grapefruit (Citrus × paradisi Macf.) peels.

Grapefruit (Citrus × paradisi Macf.) peel, a by-product of the citrus-processing industry, possesses an important economic value due to the richness of bioactive compounds. In this study, boron-linked covalent organic frameworks integrated with molecularly imprinted polymers (CMIPs) were developed via a facile one-pot bulk polymerization approach for the selective extraction of naringenin from grapefruit peel extract. The obtained CMIPs possessed a three-dimensional network structure with uniform pore size distribution, large surface areas (476 m2/g), and high crystallinity. Benefiting from the hybrid functional monomer APTES-MAA, the acylamino group can coordinate with the boronate ligands of the boroxine-based framework to form B-N bands, facilitating the integration of imprinted cavities with the aromatic skeleton. The composite materials exhibited a high adsorption capacity of 153.65 mg/g, and a short adsorption equilibrium time of 30 min for naringenin, together with favorable selectivity towards other flavonoid analogues. Additionally, the CMIPs captured the template molecules through π-π* interaction and hydrogen bonding, as verified by FT-IR and XPS. Furthermore, they had good performance when employed to enrich naringenin in grapefruit peels extract compared with the common adsorbent materials including AB-8, D101, cationic exchange resin, and active carbon. This research highlights the potential of CMIPs composite materials as a promising alternative adsorbent for naringenin extraction from grapefruit peel.

Overexpression of IFIT1 protects against LPS-induced acute lung injury via regulating CCL5-p65NF-κB signaling.

Acute lung injury (ALI) is featured by intensive inflammatory responses causing significant morbidity and mortality. Interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), induced by interferon (IFN), has been discovered to modulate viral infection and cell apoptosis and inhibit the production of pro-inflammatory cytokines. However, it's role and mechanism in ALI remain unclear and need to be explored furtherly. Here, we discovered that IFIT1 decreased the expression of TNF-α, IL-1ß and IL-6 in mouse-derived macrophage cells (MH-S) and alleviated apoptosis of murine lung epithelial cells (MLE-12) induced by MH-S cell supernatant, contributing to anti-inflammatory and antiapoptotic effects in vitro and in vivo. Moreover, RNA sequencing analysis (RNA-seq) showed that inflammatory chemokine CC motif chemokine ligand 5 (CCL5) partially eliminated the protective effects of IFIT1 and promoted the expression of inflammatory cytokines TNF-α, IL-1ß and IL-6 by CCL5-p65NF-κB signaling pathway. This study demonstrated that IFIT1 attenuated ALI-associated inflammation and cell apoptosis by regulating the CCL5-p65NF-κB signaling pathway. These findings are of great significance for the treatment of lung injury.

Algal polysaccharides and derivatives as potential therapeutics for obesity and related metabolic diseases.

The worldwide upward trend in obesity has been dramatic, but there is a lack of effective and safe drug treatment. Marine-derived algal polysaccharides, including fucoidan, alginate, polysaccharide of Spirulina platensis (PSP), ulvan, rhamnan sulfate (RS), laminarin, agar, and carrageenan, are widely used in combination with wound healing, drug delivery, and tissue-related biomedical research engineering. Recently, the prebiotic effects of algal polysaccharides and their related derivatives have received more and more attention. In this review, we discuss the potential and challenges of algal polysaccharides and their derivatives as potential therapeutic agents for obesity and its related metabolic diseases. Relevant studies have demonstrated that a variety of algal polysaccharides can play a significant role in weight loss and treatment of obesity-related metabolic diseases by improving the composition of gut microbiota, promoting bile acid formation, and upregulating cholesterol receptors in the liver. Because of their low price, non-toxic properties, and easy availability, algal polysaccharides have the potential to be developed as weight loss products.

Optimal Treatments for NSCLC Patients Harboring Primary or Acquired MET Amplification.

Background: In non-small cell lung cancer (NSCLC) patients harboring MET mutations, MET-tyrosine kinase inhibitors (TKIs) have been proven to achieve a good response. However, the relative efficacy of different therapeutics in primary NSCLC patients with MET amplification and the treatment options for patients harboring acquired MET amplification after the failure of epidermal growth factor receptor (EGFR)-TKIs remain unclear. Methods: In total, 33 patients harboring primary MET amplification and 9 patients harboring acquired MET alterations identified by next-generation sequencing were enrolled. A retrospective analysis was conducted to compare the efficacy of different therapeutics. In addition, studies reporting various treatments for patients harboring MET alterations were included in the meta-analysis. Results: In our cohort of patients harboring primary MET amplification, crizotinib displayed better efficacy than immunotherapy and chemotherapy, as demonstrated both in first-line (P = .0378) and second-line treatment regimens (P = .0181). The disease control rates for crizotinib, immunotherapy, and chemotherapy were 81.8%, 72.7%, and 63.6%, respectively. In particular, the median progression-free survival (PFS) time after immunotherapy in patients harboring MET amplification and high programed death ligand 1 (PD-L1) expression (>50%) was only 77.5 days. The meta-analysis revealed that the median PFS times after crizotinib and immunotherapy were 4.57 and 2.94 months, respectively. In patients harboring acquired MET amplification, chemotherapy plus bevacizumab had superior efficacy (310.0 days vs 73.5 days, P = .0360) compared with MET-TKIs ± EGFR-TKIs. Conclusions: Immunotherapy showed a low response in patients harboring MET alterations, even those with concurrent high PD-L1 expression. MET-TKIs might be an optional treatment with worth-expecting efficacy. However, chemotherapy plus bevacizumab could benefit the subpopulation of patients harboring acquired MET amplification after the failure of EGFR-TKIs.

A Nanopore Sensing Assay Resolves Cascade Reactions in a Multienzyme System.

Enzymatic cascade reactions are widely used to synthesize complex molecules from simple precursors. The major underlying mechanism of cascade reactions is substrate channelling, where intermediates of different enzymatic steps are not in equilibrium with the bulk solution. Here, we report a nanopore sensing assay that allows accurate quantification of all the reaction intermediates and the product of an artificial three-enzyme system. A DNA-peptide complex is used as the initial substrate which undergoes sequential enzymatic cleavages in solution. All the temporal changes of the intermediates and product can be obtained through nanopore translocation recordings. Furthermore, we find that in a confined environment such as liposome, substrate channelling occurs between two sets of the three enzymes. Our results demonstrate a novel and powerful approach to determine and quantify substrate channelling effects, which is potentially useful for designing and evaluating multienzyme systems.

High-fidelity biosensing of dNTPs and nucleic acids by controllable subnanometer channel PaMscS.

Current tools for dNTP analysis mainly rely on expensive fluorescent labeling, mass spectrometry or electrochemistry. Single-molecule assay by protein nanopores with an internal diameter of ca. 1-3.6 nm provides a useful tool for dNTP sensing. However, the most commonly used protein nanopores require additional modifications to enable dNTP detection. In this study, the PaMscS channel (mechanosensitive channel of small conductance from Pseudomonas aeruginosa) embedded in the bilayer lipid membrane (BLM) of E. coli polar lipid extract was applied as a nanopore for single molecular sensing. Two mutants of PaMscS nanopores on the side portal region (PaMscS W130A and PaMscS K180R) were selected for direct dNTP or pyrophosphoric acid (PPi) detection without aptamer or protein modification. Notably, the PaMscS mutant pore can be adjusted by regulation of osmolarity differences, which is crucial for the optimal detection of specific molecules. In addition, we established a PaMscS-based diagnosis method for the rapid sensing of disease-associated nucleic acids by monitoring the consumption of dNTPs, with 86% specificity and 100% sensitivity among 22 clinical samples. This protein nanopore, without aptamer or modification, paves a new way for dNTPs, PPi direct sensing and nucleic acid detection with low cost but high versatility.

A portable wireless intelligent electrochemical sensor based on layer-by-layer sandwiched nanohybrid for terbutaline in meat products.

The aim of this study is to develop a portable wireless intelligent nanosensor (PWIN) for rapid cost-effective on-site determination of terbutaline (TRA) residue in meat products outdoors in comparison with traditional nanosensor and high-performance liquid chromatography (HPLC). The layer-by-layer sandwiched nanohybrid fabricated by platinum-palladium nanoparticles, carboxylated graphene and graphene-like molybdenum disulfide displayed a wide linear range of 0.55-14.9 µmol/L using the portable potentiostat with smartphone, and the result was almost close to the linear range (0.4-14 µmol/L) using the traditional potentiostat with desktop computer for TRA. The limit of detections were identified as 0.44 µmol/L and 0.18 µmol/L, respectively. PWIN displayed satisfactory recovery (91%-98.43%) of TRA in samples by the standard addition method and in comparison with both traditional sensor (93.79%-98%) and HPLC (93.4%-98.6%), revealing that PWIN for rapid cost-effective on-site analysis in the food safety field is feasible.

Serum mBDNF and ProBDNF Expression Levels as Diagnosis Clue for Early Stage Parkinson's Disease.

Parkinson's disease (PD) is one of the most common chronic, progressive, and neurodegenerative diseases characterized clinically by resting tremor, bradykinesia, rigidity, and postural instability. As this disease is usually detected in the later stages, the cure is often delayed, ultimately leading to disability due to the lack of early diagnostic techniques. Therefore, it is of great importance to identify reliable biomarkers with high sensitivity and specificity for the early diagnosis of PD. In this study, we aimed to investigate whether serum expressions of mature brain-derived neurotrophic factor (mBDNF) and proBDNF can serve as biomarkers for the diagnosis of PD at early stage. One hundred and fifty-six patients with limb tremor and/or bradykinesia meeting the inclusion criteria were assigned to either ex-PD group (PD cases) or ex-NPD group (non-PD cases) and then reassigned to either po-PD group (with PD) or po-NPD group (without PD) at 1-year follow-up based on the results of the rediagnoses as performed in accordance with MDS Parkinson's diagnostic criteria. To improve early diagnostic accuracy, grouping (PD group and non-PD group) at initial visit and follow-up was performed differently and independently. Serum mBDNF and proBDNF levels were measured by enzyme-linked immunosorbent assays. The results demonstrated that serum levels of mBDNF and mBDNF/proBDNF were significantly lower in the ex-PD group (19.73 ± 7.31 and 0.09 ± 0.05 ng/ml) as compared with the ex-NPD group (23.47 ± 8.21 and 0.15 ± 0.12 ng/ml) (p < 0.01 for both) and in the po-PD group (19.24 ± 7.20 and 0.09 ± 0.05 ng/ml) as compared with the po-NPD group (25.05 ± 7.67 and 0.16 ± 0.14 ng/ml) (p < 0.01 for both). However, a significantly higher serum level of proBDNF was noted in the ex-PD group (235.49 ± 60.75 ng/ml) as compared with the ex-NPD group (191.75 ± 66.12 ng/ml) (p < 0.01) and in the po-PD group (235.56 ± 60.80 ng/ml) as compared with the po-NPD group (188.42 ± 65.08 ng/ml) (p < 0.01). In conclusion, mBDNF/proBDNF can be used as biomarkers for early stage Parkinson's disease; in addition, mBDNF plus proBDNF has better diagnostic value than mBDNF alone in the diagnosis of PD.

Portable wireless intelligent sensing of ultra-trace phytoregulator α-naphthalene acetic acid using self-assembled phosphorene/Ti3C2-MXene nanohybrid with high ambient stability on laser induced porous graphene as nanozyme flexible electrode.

The harm of pesticide residues to human health via environmental pollution in agriculture has recently become a significant livelihood issue. Herein, a new strategy for smart ultra-trace analysis of phytoregulator α-naphthalene acetic acid (NAA) residues in farmland environments and agro-products via machine learning (ML) using a nanozyme flexible electrode fabricated by two-dimensional phosphorene (BP) nanohybrid with graphene-like titanium carbide MXene (Ti3C2-MXene) on the flexible substrate surface of laser-induced porous graphene (LIPG) is proposed. Highly ambient-stable BP nanohybrid with Ti3C2-MXene is prepared by ultrasonic-assisted liquid-phase exfoliation in organic solvent containing grinding black phosphorus, cuprous chloride and, Ti3C2-MXene that is obtained by selectively etching Al layers of Ti3AlC2. Nanozyme flexible electrode is fabricated by drop-coating Ti3C2-MXene/BP that is formed through electrostatic self-assembly between positively charged BP and negatively charged Ti3C2-MXene onto LIPG that is obtained by direct laser writing on commercial polyimide and patterned via a computer-aided design system as a flexible substrate. The ML model via artificial neural network algorithm for smart output of NAA is discussed. NAA is electrochemically detected in a wide linear range of 0.02-40 µM with a low limit of detection (LOD) of 1.6 nM using a portable mini-workstation. Large and rough surfaces, excellent electrochemical response, and satisfactory practicability demonstrated the feasibility and detectability of the proposed method. This will provide a portable wireless intelligent nanozyme flexible sensing platform for cost-effective, simple, fast and, ultra-trace detection of hazardous substances in the safety of environments, products, and food in agriculture.

A highly-sensitive and selective antibody-like sensor based on molecularly imprinted poly(L-arginine) on COOH-MWCNTs for electrochemical recognition and detection of deoxynivalenol.

A new strategy to mimic antibody for electrochemical recognition and detection of deoxynivalenol (DON) using a highly-sensitive and selective antibody-like sensor based on molecularly imprinted poly(l-arginine) (P-Arg-MIP) on carboxylic acid functionalized carbon nanotubes (COOH-MWCNTs) was proposed. l-arginine as functional monomer was screened to prepare imprinted electrode via its electro-polymerization in the presence of DON onto the surface of COOH-MWCNTs electrode coupled with theoretical calculation. Surface morphology, structural characteristics, and electrochemical properties of P-Arg-MIP/COOH-MWCNTs were characterized by SEM, EDS, FTIR, and CV, respectively. P-Arg-MIP/COOH-MWCNTs displayed relatively high conductivity, high effective surface area, antibody-like molecular recognition and affinity, and a good response towards DON in a linear range from 0.1 to 70 µM with LOD of 0.07 µM in wheat flour samples with satisfactory recovery and feasible practicability in comparison with HPLC. This method provides a promising biomimetic sensing platform for the determination of mycotoxins in food and agro-products.

Research on Unsafe Behavior of Construction Workers Under the Bidirectional Effect of Formal Rule Awareness and Conformity Mentality.

At present, China's engineering safety management has developed to a certain level, but the number of casualties caused by construction accidents is still increasing in recent years, and the safety problems in the construction industry are still worrying. For purpose of effectively reducing construction workers' unsafe behavior and improve the efficiency of construction safety management, based on multi-agent modeling, this paper analyzes the influencing factors during construction workers' cognitive process from the perspective of safety cognition, constructs the interaction and cognition of the agent under the bidirectional effect of formal rule awareness and conformity mentality model, and set behavior rules and parameters through the Net Logo platform for simulation. The results show that: Unsafe behavior of construction workers is related to the failure of cognitive process, and the role of workers' psychology and consciousness will affect the cognitive process; The higher the level of conformity intention of construction workers, the easier it is to increase the unsafe behavior of the group; Formal rule awareness can play a greater role only when the management standard is at a high level, and can correct the workers' safety cognition and effectively correct the workers' unsafe behavior; Under certain construction site environmental risks, the interaction between formal rule awareness and conformity mentality in an appropriate range is conducive to the realization of construction project life cycle management. This study has certain theoretical and practical significance for in-depth understanding of safety cognition and reducing unsafe behavior of construction team.

Probing Conformational Polymorphism of DNA Assemblies with Nanopores.

Single-stranded DNA (ssDNA) can be designed to assemble into duplexes and other high-order structures through Watson-Crick hydrogen bonds. Incorporation of unnatural nucleobases or binding with small molecules can also introduce new interactions that give rise to novel DNA assemblies. However, the methods for determining the conformational properties of DNA assemblies are still very limited. Here we develop a new strategy for probing conformational polymorphism of different DNA assemblies. By installing poly(dC)30 tails to the ends of individual ssDNA that assemble into duplex, triplex, or other complex structures, we are able to observe different current blockade patterns corresponding to specific DNA nanostructures when the DNA assemblies are lodged inside α-hemolysin vestibule. We can also monitor the disassembly of the DNA nanostructures in solution. This method complements the existing traditional technologies such as circular dichroism spectroscopy, fluorescence labeling, and NMR spectroscopy, and shows distinct advantages of high accuracy and general applicability.

Measuring Enzymatic Activities with Nanopores.

Nanopores have become powerful and versatile tools for measuring single molecules since their emergence in the mid-1990s. They can be used to sense a wide variety of analytes including metal ions, small organic molecules, DNA/RNA, proteins, etc. to monitor chemical reactions, and to sequence DNA. Recently, enzymes have been studied by using nanopore technologies. In this Minireview, we highlight recent efforts in developing nanopore enzymology and categorize the related work into three groups: 1) measuring enzymatic activities with nanopore-enzyme hybrids; 2) measuring enzymatic activities through sensing their catalytic products with nanopores; 3) the use of enzymes for DNA sequencing and DNA/protein translocation. At the end, we discuss the challenges and opportunities in nanopore enzymology.

In-situ reduction of Ag+ on black phosphorene and its NH2-MWCNT nanohybrid with high stability and dispersibility as nanozyme sensor for three ATP metabolites.

The environmental stability, water-processibility and life-span of black phosphorene (BP) severely limit the application of its electronic devices in aqueous system containing oxygen. We reported the controllable preparation of in-situ reduction and deposition of silver nanoparticles on the BP surface and its amino-functionalized multi-walled carbon nanotubes (NH2-MWCNT) nanocomposite. With the addition of both NH2-MWCNT and Ag+, the BP-based nanocomposite was prepared by ultrasonic-assisted liquid-phase exfoliation and was dispersed in carboxymethyl cellulose sodium (CMC) aqueous solution. The morphology, microstructure, and electrochemical properties of the nanohybrid were characterized. NH2-MWCNT-BP-AgNPs showed high environmental stability, good water-processibility, satisfactory life-spans, superior electrocatalytic capacity with enzyme-like kinetic characteristics. The nanohybrid was applied as electrochemical sensors for single/simultaneous analysis of uric acid (UA), xanthine (XT) and hypoxanthine (HX). Excellent voltammetric responses for simultaneous determination in linear ranges of 0.1-800 µM with a limit of detection (LOD) of 0.052 µM for UA, 0.5-680 µM with a LOD of 0.021 µM for XT, and 0.7-320 µM with a LOD of 0.025 µM for HX under optimal conditions. Besides, the developed nanozyme sensor was employed for simultaneous voltammetric analysis of UA, XT and HX in real samples with acceptable recoveries. This work will provide theoretical guidance and experimental support for the preparation and application of two-dimensional materials, nanozymes and sensing devices.

Electrochemical detection combined with machine learning for intelligent sensing of maleic hydrazide by using carboxylated PEDOT modified with copper nanoparticles.

A method for intelligent data analysis was designed by combining electrochemical sensing with machine learning (ML). Specifically, a voltammetric sensor is described for determination of the phytoinhibitor maleic hydrazide in crop samples. Carboxyl-functionalized poly(3,4-ethylenedioxythiophene) (PEDOT-C4-COOH) was electro-synthesized in aqueous micellar solution by direct anodic oxidation of its monomer. A nanosensor was then prepared by placing copper nanoparticles (CuNPs) on the PEDOT-C4-COOH film via electro-deposition of Cu (II) from aqueous micellar solutions. An artificial neural network (ANN) served as a powerful ML model to realize intelligent data analysis and smart transformation for digital output. Different established regression methods were selected for evaluating the ANN-based method that was found to be superior to known methods. The sensor has a wide working range (from 0.06-1000 µM), a low limit of detection (10 nM), good stability, selectivity and practicality. The method was applied to the determination of maleic hydrazide in (spiked) samples of onion, rice, potato and cotton leaf. Satisfactory results demonstrate that the feature of simultaneous data acquisition and analysis is highly attractive. Graphical abstract Schematic representation of an electrochemical sensor based on carboxyl-functionalized poly(3,4-ethylenedioxythiophene) (PEDOT-C4-COOH) and copper nanoparticles (CuNPs) by differential pulse voltammetry (DPV) to detect maleic hydrazide (MH). PEDOT-C4-COOH was electro-synthesized in 0.1 M LiClO4 aqueous micellar solution with 0.1 M sodium dodecyl benzene sulfonate (SDBS) by amperometry (CA). CuNPs was prepared by cyclic voltammetry (CV).

Enhanced Sensitivity in Nanopore Sensing of Cancer Biomarkers in Human Blood via Click Chemistry.

Cancer biomarkers are expected to be indicative of the occurrence of certain cancer diseases before the tumors form and metastasize. However, many biomarkers can only be acquired in extremely low concentrations, which are often beyond the limit of detection (LOD) of current instruments and technologies. A practical strategy for nanopore sensing of cancer biomarkers in raw human blood down to the femtomolar level is developed here. This strategy first converts the detection of cancer biomarkers to the quantification of copper ions by conducting a sandwich assay involving copper oxide nanoparticles. The released Cu2+ is then taken to catalyze the "click" reaction which ligates a host-guest modified DNA probe. Finally, this DNA probe is subjected to single-channel recordings to afford the translocation events that can be used to derive the concentrations of the original biomarkers. Due to the amplification effects of nanoparticle loadings and the "click" reaction, the LOD of this strategy can be as low as the subfemtomolar level. Further, the acid treatment step could effectively eliminate the interferences from plasma proteins in raw human blood and make the strategy highly suitable for the detection of cancer biomarkers in clinical samples.

Amplified Split Aptamer Sensor Delivered Using Block Copolymer Nanoparticles for Small Molecule Imaging in Living Cells.

We develop a novel amplified split aptamer sensor for highly sensitive detection and imaging of small molecules in living cells by using cationic block copolymer nanoparticles (BCNs) with entrapped fluorescent conjugated polymer as a delivery agent. The design of a split aptamer as the initiator of hybridization chain reaction (HCR) affords the possibility of enhancing the signal-to-background ratio and thus allows high-contrast imaging for small molecules with relatively weak interactions with their aptamers. The novel design of using fluorescent cationic BCNs as the nanocarrier enables efficient and self-tracking transfection of DNA probes. Results reveal that BCNs exhibit high fluorescence brightness allowing direct tracking of the delivery location. The developed amplified split aptamer sensor is shown to have high sensitivity and selectivity for in vitro quantitative detection of adenosine triphosphate (ATP) with a detection limit of 30 nM. Live cell studies show that the sensor provides a "signal on" approach for specific, high-contrast imaging of ATP. The DNA sensor based HCR system may provide a new generally applicable platform for detection and imaging of low-abundance biomarkers.

Multiplexed discrimination of microRNA single nucleotide variants through triplex molecular beacon sensors.

Herein, we develop a new nanopore sensing strategy for the selective detection of microRNAs and single nucleotide variants (SNVs) based on triplex molecular beacon sensors. This sensing system shows very high specificity in discriminating microRNA SNVs and can be applied for the simultaneous detection of several microRNAs of the same family in a mixture.

Rapid and selective DNA-based detection of melamine using α-hemolysin nanopores.

We have developed a rapid and selective approach for the detection of melamine based on simple DNA probes and α-hemolysin nanopores. The limit of detection can be as low as 10 pM.

Analyte-Triggered DNA-Probe Release from a Triplex Molecular Beacon for Nanopore Sensing.

A new nanopore sensing strategy based on triplex molecular beacon was developed for the detection of specific DNA or multivalent proteins. The sensor is composed of a triplex-forming molecular beacon and a stem-forming DNA component that is modified with a host-guest complex. Upon target DNA hybridizing with the molecular beacon loop or multivalent proteins binding to the recognition elements on the stem, the DNA probe is released and produces highly characteristic current signals when translocated through α-hemolysin. The frequency of current signatures can be used to quantify the concentrations of the target molecules. This sensing approach provides a simple, quick, and modular tool for the detection of specific macromolecules with high sensitivity and excellent selectivity. It may find useful applications in point-of-care diagnostics with a portable nanopore kit in the future.