A covalent organic framework nanosheet-nanochannel composite with signal amplification strategy for electrochemical enantioselective recognition.

Recognition and separation of chiral isomers are of great importance in both industrial and biological applications. However, owing to identical molecular formulas and chemical properties of enantiomers, signal transduction and amplification are still two major challenges in chiral sensing. In this study, we developed an enantioselective device by integrating chiral covalent organic framework nanosheets (CONs) with nanochannels for sensitive identification and quantification of enantiomers. Using 3,4-dihydroxyphenylalanine (DOPA) as the model analyte, the as-prepared chiral nanofluidic device exhibits a remarkable chiral recognition ability to l-DOPA than d-DOPA. More importantly, due to the chelation of DOPA with Fe3+ ions, it can efficiently block the ion transport through channel and shield the channel surface charge, which will amplify the difference in the electrochemical response of l-DOPA and d-DOPA. Therefore, a sensitive chiral recognition can be achieved using the present nanofluidic device coupled using electrochemical amplification strategy. Notably, using this method, an ultra-low concentration of l-DOPA (as low as 0.21 pM) can be facilely and successfully detected with a linear range of 1 pM-10 µM. This study provides a reliable and sensitive approach for achieving highly selective detection of chiral molecules.

Integrated separation and detection of exosomes via a label-free magnetic SERS platform.

A label-free magnetic surface enhanced Raman scattering (SERS) platform was constructed, which was composed of superparamagnetic Fe3O4 nanoparticles as cores for separation and Au layers as shells for label-free SERS detection. Our method could effectively distinguish exosomes from different cell sources for cancer diagnosis and showed high sensitivity and specificity within a 95% confidence interval. As a low-cost and efficient exosome analysis method, the designed integrated platform for separation and detection has promising applicability in clinical diagnostics.

Nanozyme-catalyzed cascade reaction enables a highly sensitive detection of live bacteria.

The accurate and timely detection of bacteria is critically important for human health as it helps to determine the original source of bacterial infections and prevent disease spread. Herein, gold nanoparticles (AuNPs) were synthesized using polyoxometalates (POMs) as the stabilizing agent. Since AuNPs have glucose oxidase (GOx)-like activity and POMs possess peroxidase (HRP)-like activity, the as-prepared Au@POM nanoparticles have double enzyme-like activities and facilitate cascade reaction. As known, glucose is required as an energy resource during bacterial metabolism, the concentration of glucose decreases with the increase of bacteria content in a system with bacteria and glucose. Therefore, when we use Au@POM nanozymes to trigger the cascade catalysis of glucose and 3,3',5,5'-tetramethylbenzidine (TMB), the concentration of glucose and bacteria can be sensitively detected using the absorbance intensity at 652 nm in the visible spectrum. As demonstration, S. aureus and E. coli were used as model bacteria. The experimental results show that the present method has a good linear relationship in the bacterial concentration range of 1 to 7.5 × 107 colony-forming units (CFU) mL-1 with a detection limit of 5 CFU mL-1. This study shows a great promise of nanozyme cascade reactions in the construction of biosensors and clinical detections.

Simultaneous Sensing of Multiplex Volatile Organic Compounds by Adsorption and Plasmon Dual-Induced Raman Enhancement Technique.

Developing highly efficient gas sensors with excellent performance for rapid and sensitive detection of volatile organic compounds (VOCs) is of critical importance for the protection of human health, ecological environment, and other factors. Here, a robust gas sensor based on Raman technology was constructed by an in situ grown 2D covalent organic framework (COF) on Au nanoparticles' surface in the microchannel. Dual enhancement effects are included for the as-prepared microfluidic sensor. First, acting as a gas confinement chamber, the 2D COF could effectively capture gas molecules with high adsorption capacity and fast adsorption kinetics, resulting in VOCs' preconcentration at a high level in the COF layer. At the same time, after being stacked in the microchannel, abundant hot spots were generated among the nanogaps of Au@COF NPs. The local surface plasmon resonance effect could effectively enhance the Raman intensity. Both factors contribute to the improved detection sensitivity of VOCs. As a demonstration, several representative VOCs with different functional groups were tested. The resultant Raman spectra were subjected to the statistical principal component analysis. Varied VOCs can be successfully detected with a detection limit as low as ppb level and distinguished with 95% confidence interval. The present microfluidic platform provides a simple, sensitive, and fast method for VOCs' sensing and distinguishing, which is expected to hold potential applications in the fields of health, agricultural, and environmental research.

Composition of major quinochalcone hydroxysafflor yellow A and anhydrosafflor yellow B is associated with colour of safflower (Carthamus tinctorius) during colour-transition but not with overall antioxidant capacity: A study on 144 cultivars.

Yellow pigments in the water-extract of safflower (Carthamus tinctorius L.) belong to quinochalcone flavonoid family and are widely used as food colourants. The aim of the study was to characterize the main quinochalcone compounds in safflower water-extract during blooming period when floret changed colour. Mass-spectrometry results showed that hydroxysafflor yellow A (HSYA) and anhydrosafflor yellow B (AHSYB) were the most abundant. Based on 370 florets samples collected from 144 cultivars, the contents of HSYA and AHSYB were determined, which showed that only AHSYB content had relatively strong positive association with colour indexes. The ratio of HSYA/AHSYB and visual colour exhibited certain patterns: yellow = 2, orange = 3-4, red = more dispersed, mostly falling 5-6. Most of the florets had HSYA increased first and decreased, while AHSYB decreased all the time when floret changed colour as yellow â†’ orange â†’ red. Regardless of the composition of HSYA/AHSYB in florets, the antioxidant capacities of safflower petal water-extracts were the same.

Plasmonic Nanozymes: Localized Surface Plasmonic Resonance Regulates Reaction Kinetics and Antibacterial Performance.

Among the members of the rapidly growing nanozyme family, plasmonic nanozymes stand out because of their unique localized surface plasmon resonance (LSPR) characteristics and tunable catalytic activity. We prepared a plasmonic nanozyme of Au gold nanoparticles (AuNPs) and Cu metal-organic framework nanosheets (Cu-MOFNs). The Cu-MOFNs have peroxidase-like activity, while AuNPs present unique LSPR characteristics. We found that the as-prepared AuNPs/Cu-MOFNs composite presents 1.6-fold faster reaction kinetics under LSPR excitation compared to that in the dark. Investigations of energy levels, radical capture, and dark-field scattering spectroscopy revealed that LSPR of AuNPs as well as matched energy levels can facilitate efficient hot electron transfer, which could readily cleave the chemical bond of the substrate and accelerate the reaction kinetics. On the basis of these results, we achieved enhanced antibacterial therapy and wound healing using plasmonic AuNPs/Cu-MOFNs. This study spotlights the superiority of plasmonic nanozymes in improving the enzyme-like performance of nanozymes.

Ultrasensitive and Label-Free Detection of Cell Surface Glycan Using Nanochannel-Ionchannel Hybrid Coupled with Electrochemical Detector.

In this work, asymmetric nanochannel-ionchannel of porous anodic alumina (PAA) coupled with electrochemical detector was used for sensitive and label-free detection of cell surface glycan. The amplified ionic current caused by array nanochannels as well as the ionic current rectification (ICR) caused by asymmetric geometry endows PAA with sensitive ionic current response. Functionalized with the special molecular probe, the constructed nanofluidic device can be used for selective recognition and detection of glycan in a real-time and label-free format. In addition, due to the subnanosize of ionchannels, the probe immobilization and glycan recognition is carried out on the outer surface of PAA, avoiding the blockage of PAA nanochannel by samples, which promises the reproducibility and accuracy of the present method toward bioanalysis. Results show that the glycan concentration ranging from 10 fM to 10 nM can be successfully detected with a detection limit of ∼10 aM, which is substantially lower than most previous works. The designed strategy provides a valuable platform for sensitive and label-free detection of cell surface glycan, which acts as a promising candidate in pathological research and cancer diagnosis.

Retraction Note: A switch in the poly(dC)/RmlB complex regulates bacterial persister formation.

This paper has been retracted.

A switch in the poly(dC)/RmlB complex regulates bacterial persister formation.

Bacterial persisters are phenotypic variants that tolerate exposure to lethal antibiotics. These dormant cells are responsible for chronic and recurrent infections. Multiple mechanisms have been linked to persister formation. Here, we report that a complex, consisting of an extracellular poly(dC) and its membrane-associated binding protein RmlB, appears to be associated with persistence of the opportunistic pathogen Pseudomonas aeruginosa. Environmental stimuli triggers a switch in the complex physiological state (from poly(dC)/RmlB to P-poly(dC)/RmlB or RmlB). In response to the switch, bacteria decrease proton motive force and intracellular ATP levels, forming dormant cells. This alteration in complex status is linked to a (p)ppGpp-controlled signaling pathway that includes inorganic polyphosphate, Lon protease, exonuclease VII (XseA/XseB), and the type III secretion system. The persistence might be also an adaptive response to the lethal action of the dTDP-L-rhamnose pathway shutdown, which occurs due to switching of poly(dC)/RmlB.

Transfer of a cyanobacterial neurotoxin, ß-methylamino-l-alanine from soil to crop and its bioaccumulation in Chinese cabbage.

Most cyanobacteria can synthesize the notorious neurotoxin ß-methylamino-l-alanine (BMAA) that is transferred and bioaccumulated through natural food webs of aquatic ecosystems and ultimately arises the potential human health risks by the consumption of BMAA-contaminated aquatic products. Fertilization of cyanobacterial composts in farmlands may also lead to BMAA contamination in soil and its possible transfer and bioaccumulation within major crops, thereby threatening human health. In this study, we first detected a high level of BMAA (1.8-16.3 µg g-1) in cyanobacterial composts. In order to assess the health risks from cyanobacterial composts, we planted Chinese cabbage, a favourite vegetable in China, in BMAA-contaminated soil (4.0 µg BMAA/g soil) and detected the levels of free and protein-associated BMAA in soil and crop organs during the whole growth cycle by HPLC-MS/MS, respectively. Our results demonstrated that BMAA indeed transferred from soil to root, stem and leaf of Chinese cabbage during the growth cycle. The BMAA level finally accumulated in the edible portions was much higher than the initial level in soil, including 13.82 µg g-1 in leaf and 4.71 µg g-1 in stem. The discovery of the neurotoxin BMAA in this vegetable, an ending in human consumption, not only provides a BMAA transfer pathway in farmland ecosystems, but also is alarming and requires attention due to the potential risk of cyanobacterial composts to human health.

Occurrence and transfer of a cyanobacterial neurotoxin ß-methylamino-L-alanine within the aquatic food webs of Gonghu Bay (Lake Taihu, China) to evaluate the potential human health risk.

To evaluate the health risk of cyanobacterial blooms, the levels of the neurotoxic non-protein amino acid, ß-methylamino-l-alanine (BMAA), was investigated in the freshwater ecosystem of Gonghu Bay in Lake Taihu. Lake Taihu is a large shallow lake contaminated by the excessive growth of Microcystis. Since BMAA has been measured in diverse cyanobacteria in different ecosystems all over the world, BMAA might also occur in Gonghu Bay. A long term monitoring of BMAA was done by HPLC-MS/MS method in cyanobacteria, mollusks, crustaceans and various fish species at different trophic levels of ecosystems in Gonghu Bay, some of which were popularly consumed by humans. Over the entire sampling period, the total average BMAA content in cyanobacteria, mollusks, crustaceans and various fish species were 4.12, 3.21, 3.76, and 6.05µgBMAA/g dry weight, respectively. Thus, BMAA could be biosynthesized by the blooming cyanobacteria in which Microcystis dominates. This toxin can be transferred through ascending trophic levels of the aquatic ecosystem in Gonghu Bay. The bioaccumulation of BMAA was observed in aquatic animals, especially in some fish species during the bloom-outbreak and bloom-decline phases. The discovery of the chronic neurotoxin BMAA in a large limnic ecosystem together with possible pathways of accumulation within major food webs deserves serious consideration due to its potential long-term risk to human health.

Identification and characterization of a cold-active phthalate esters hydrolase by screening a metagenomic library derived from biofilms of a wastewater treatment plant.

A cold-active phthalate esters hydrolase gene (designated dphB) was identified through functional screening of a metagenomic library derived from biofilms of a wastewater treatment plant. The enzyme specifically catalyzed the hydrolysis of dipropyl phthalate, dibutyl phthalate, and dipentyl phthalate to the corresponding monoalkyl phthalate esters at low temperatures. The catalytic triad residues of DphB were proposed to be Ser159, Asp251, and His281.

Extension of Sphingobium sp. BHC-A to a 2,4,5-trichlorophenoxyacetic acid mineralizing strain by metabolic engineering.

The gene cassette encoding for TftAB and TftCD proteins was integrated into the 16srDNA gene of the γ-hexachlorocyclohexane (γ-HCH) mineralizing strain Sphingobium sp. BHC-A by homologous recombination. The recombinant γ-HCH mineralizing strain may degrade 2,4,5-trichlorophenoxyacetic acid at a rate of 250nmol mg [protein](-1)h(-1), and the generated intermediate 2,5-dichlorohydroquinone may be further mineralized through γ-HCH downstream degradation pathway.

Cloning of a dibutyl phthalate hydrolase gene from Acinetobacter sp. strain M673 and functional analysis of its expression product in Escherichia coli.

A dibutyl phthalate (DBP) transforming bacterium, strain M673, was isolated and identified as Acinetobacter sp. This strain could not grow on dialkyl phthalates, including dimethyl, diethyl, dipropyl, dibutyl, dipentyl, dihexyl, di(2-ethylhexyl), di-n-octyl, and dinonyl phthalate, but suspensions of cells could transform these compounds to phthalate via corresponding monoalkyl phthalates. During growth in Luria-Bertani medium, M673 produced the high amounts of non-DBP-induced intracellular hydrolase in the stationary phase. One DBP hydrolase gene containing an open reading frame of 1,095 bp was screened from a genomic library, and its expression product hydrolyzed various dialkyl phthalates to the corresponding monoalkyl phthalates.

An improved particle bombardment for the generation of transgenic plants by direct immobilization of relleasable Tn5 transposases onto gold particles.

We have developed a modified particle bombardment method for plant transgenesis. An intein-tag and a 6×Cys-tag were successively fused to the N-terminus of a hyperactive Tn5 transposase. The modified transposase was immobilized on bare gold microscopic particles via covalent binding of a 6×Cys-tag sulfydryl groups to the gold surface. The tethered transposase can bind the transposon DNA in vitro to form the transposome in the absence of Mg²âº ions. After bombardment of the gold particles carrying the transposomes into the plant cells, the transposomes will be released from the carrier due to the activated self-cleavage function of intein-tag. Our data showed this procedure integrated foreign DNA into the plant genome with an increased transformation frequency as compared to the conventional particle bombardment method. A single copy insertion can also be obtained by decreasing of the assembled transposon DNA amount in relation to plant cell biomass.

Tn5 transposase-assisted transformation of indica rice.

Here, we describe experiments on Tn5 transposase-assisted transformation of indica rice. Transposomes were formed in vitro as a result of hyperactive Tn5 transposase complexing with a transposon that contained a 19-bp tetracycline operator (tetO) sequence. To form modified projectiles for transformation, the Tn10-derived prokaryotic tetracycline repressor (TetR) proteins, which can bind transposomes via the high affinity of TetR for tetO, were immobilized onto the surface of bare gold microscopic particles. These projectiles were introduced into cells of the indica rice cultivar Zhuxian B by particle bombardment. Once projectiles were inside the cell, tetracycline induced an allosteric conformational change in TetR that resulted in the dissociation of TetR from tetO, and thus generated free transposomes. Molecular evidence of transposition was obtained by the cloning of insertion sites from many transgenic plants. We also demonstrated that the introduced foreign DNA was inherited stably over several generations. This technique is a promising transformation method for other plant species as it is species independent.

A linear modulation-based stochastic resonance algorithm applied to the detection of weak chromatographic peaks.

A simple stochastic resonance algorithm based on linear modulation was developed to amplify and detect weak chromatographic peaks. The output chromatographic peak is often distorted when using the traditional stochastic resonance algorithm due to the presence of high levels of noise. In the new algorithm, a linear modulated double-well potential is introduced to correct for the distortion of the output peak. Method parameter selection is convenient and intuitive for linear modulation. In order to achieve a better signal-to-noise ratio for the output signal, the performance of two-layer stochastic resonance was evaluated by comparing it with wavelet-based stochastic resonance. The proposed algorithm was applied to the quantitative analysis of dimethyl sulfide and the determination of chloramphenicol residues in milk, and the good linearity of the method demonstrated that it is an effective tool for detecting weak chromatographic peaks.