Accelerating the Phosphatase-like Activity of Uio-66-NH2 by Catalytically Inactive Metal Ions and Its Application for Improved Fluorescence Detection of Cardiac Troponin I.

Compared with natural enzymes, nanozymes usually exhibit much lower catalytic activities, which limit the sensitivities of nanozyme-based immunoassays. Herein, several metal ions without enzyme-like activities were engineered onto Uio-66-NH2 nanozyme through postsynthetic modification. The obtained Mn+@Uio-66-NH2 (Mn+ = Zn2+, Cd2+, Co2+, Ca2+and Ni2+) exhibited improved phosphatase-like catalytic activities. In particular, a 12-fold increase in the catalytic efficiency (kcat/Km) of Uio-66-NH2 was observed after the modification with Zn2+. Mechanism investigations indicate that both the amino groups and oxygen-containing functional groups in Uio-66-NH2 are the binding sites of Zn2+, and the modified Zn2+ ions on Uio-66-NH2 serve as the additional catalytic sites for improving the catalytic performance. Furthermore, the highly active Zn2+@Uio-66-NH2 was used as a nanozyme label to develop a fluorescence immunoassay method for the detection of cardiac troponin I (cTnI). Compared with pristine Uio-66-NH2, Zn2+@Uio-66-NH2 can widen the linear range by 1 order of magnitude (from 10 pg/mL-1 µg/mL to 1 pg/mL-1 µg/mL) and also lower the detection limit by 5 times (from 4.7 pg/mL to 0.9 pg/mL).

Engineering of a Self-Regulatory Bidirectional DNA Assembly Circuit for Amplified MicroRNA Imaging.

The engineering of catalytic hybridization DNA circuits represents versatile ways to orchestrate a complex flux of molecular information at the nanoscale, with potential applications in DNA-encoded biosensing, drug discovery, and therapeutics. However, the diffusive escape of intermediates and unintentional binding interactions remain an unsolved challenge. Herein, we developed a compact, yet efficient, self-regulatory assembly circuit (SAC) for achieving robust microRNA (miRNA) imaging in live cells through DNA-templated guaranteed catalytic hybridization. By integrating the toehold strand with a preblocked palindromic fragment in the stem domain, the proposed miniature SAC system allows the reactant-to-template-controlled proximal hybridization, thus facilitating the bidirectional-sustained assembly and the localization-intensified signal amplification without undesired crosstalk. With condensed components and low reactant complexity, the SAC amplifier realized high-contrast intracellular miRNA imaging. We anticipate that this simple and template-controlled design can enrich the clinical diagnosis and prognosis toolbox.

Few-layered boron nitride nanosheet as a non-metallic phosphatase nanozyme and its application in human urine phosphorus detection.

Human urine phosphorus (existing in the form of phosphate) is a biomarker for the diagnosis of several diseases such as kidney disease, hyperthyroidism, and rickets. Therefore, the selective detection of phosphate in urine samples is crucial in the field of clinical diagnosis. Herein, we reported the phosphatase-like catalytic activity of few-layered h-BNNS for the first time. As the phosphatase-like activity of few-layered h-BNNS could be effectively inhibited by phosphate, a selective fluorescent method for the detection of phosphate was proposed. The linear range for phosphate detection is 0.5-10 µM with a detection limit of 0.33 µM. The fluorescent method was then explored for the detection of human urine phosphorus in real samples. The results obtained by the proposed method were consistent with those of the traditional method, indicating that the present method has potential application for urine phosphorus detection in clinical disease diagnosis.

Design, synthesis and imaging of a novel mitochondrial fluorescent nanoprobe based on distyreneanthracene-substituted triphenylphosphonium salt.

Targeting and monitoring the dynamics of mitochondria are of great significance because mitochondria are involved in a variety of physiological and pathological processes. For achieving this purpose, highly sensitive, photostable, tolerance and specific fluorescent probe is necessary. To obtain a superior mitochondrial fluorescent probe, (4-distyreneanthracenoxybutyl) bis(triphenylphosphonium) bromide (DSA-TPP) with aggregation-induced emission (AIE) characteristic was designed and synthesized for mitochondrial targeting. DSA-TPP dots with high fluorescence quantum yield (Φ = 17.9) and small particle size (8 nm) can be easily prepared by self-assembly formation. DSA-TPP dots had the ability of lightning mitochondria in living cells with high brightness, superior photostability and strong tolerance to cell environment change.

A colorimetric heparin assay based on the inhibition of the oxidase mimicking activity of cerium oxide nanoparticles.

A colorimetric method is described for the sensitive detection of heparin (Hep). It is based on the finding that Hep can effectively inhibit the oxidase mimicking activity of cerium oxide nanoparticles (nanoceria). In the presence of Hep, the catalytic activity of nanoceria toward the oxidation of the chromogenic substrate 3,3',5,5'-tetramethylbenzidine by oxygen is strongly decreased. The inhibition mechanism is attributed to the fact that Hep is adsorbed on the surface of the nanoceria. Under optimal condition, the absorbance (measured at 652 nm) decreases with increasing Hep concentrations in the range from 30 to 700 nM. The detection limit is 20 nM. The method was applied to the determination of Hep in medical injection sample and serum sample with satisfactory results. Graphical abstract Schematic presentation of the inhibition of oxidase-like activity of nanoceria by heparin. This allows the sensitive detection of heparin in medical injection sample and serum sample with satisfactory results.

Accelerating the Peroxidase-Like Activity of Gold Nanoclusters at Neutral pH for Colorimetric Detection of Heparin and Heparinase Activity.

The peroxidase-like catalytic activity of gold nanoclusters (Au-NCs) is quite low around physiological pH, which greatly limits their biological applications. Herein, we found heparin can greatly accelerate the peroxidase-like activity of Au-NCs at neutral pH. The catalytic activity of Au-NCs toward the peroxidase substrate 3,3',5,5'-tetramethylbenzidine (TMB) oxidation by H2O2 was 25-fold increased in the presence of heparin at pH 7. The addition of heparin not only accelerated the initial catalytic rate of Au-NCs but also prevented the Au-NCs from catalyst deactivation. This allows the sensitive colorimetric detection of heparin at neutral pH. In the presence of heparinase, heparin was hydrolyzed into small fragments, weakening the enhancement effect of catalytic activity. On the basis of this phenomenon, the colorimetric determination of heparinase in the range from 0.1 to 3 µg·mL-1 was developed with a detection limit of 0.06 µg·mL-1. Finally, the detection of heparin and heparinase activity in diluted serum samples was also demonstrated.

Switchable fluorescence of MoS2 quantum dots: a multifunctional probe for sensing of chromium(VI), ascorbic acid, and alkaline phosphatase activity.

A simple strategy for modulating the fluorescence of MoS2 quantum dots (QDs) is described. The fluorescence of MoS2 QDs was firstly switched off by the addition of Cr(VI), and the quenched fluorescence was further switched on by introducing ascorbic acid (AA) into the mixture. The fluorescence quenching of MoS2 QDs by Cr(VI) was attributed to the fluorescence inner filter effect. After the addition of AA, Cr(VI) was reduced to Cr(III), and the fluorescence was restored. This finding has been applied for the fluorescent sensing of Cr(VI) in drinking water and AA in serum samples. In addition, the present method has been extended for turn-on sensing of an important biomarker alkaline phosphatase (ALP). There is a linear relationship between the fluorescence intensity and the concentrations of ALP in the range from 2.5 to 50 U/L, and the limit of detection is 0.34 U/L. The results showed MoS2 QDs hold great potential as a multifunctional fluorescent probe for the detection of metal ions, biological small molecules, and proteins. Graphical abstract The fluorescence of MoS2 QDs can be switched off by Cr(VI), and the quenched fluorescence can be further switched on by the addition of ascorbic acid or enzymatically generated ascorbic acid. This allows the selective detection of Cr(VI), ascorbic acid, and alkaline phosphatase based on the fluorescence of MoS2 QDs.

Fluorometric turn-on determination of the activity of alkaline phosphatase by using WS2 quantum dots and enzymatic cleavage of ascorbic acid 2-phosphate.

A method is described for the determination of the activity of alkaline phosphatase (ALP). It is based on the reversible modulation of the fluorescence of WS2 quantum dots (QDs). The fluorescence of the QDs is quenched by Cr(VI) but restored by free ascorbic acid (AA). The detection scheme relies on the fact that ALP hydrolyzes the substrate ascorbic acid 2-phosphate to produce AA, and that enzymatically generated AA can restore the fluorescence of the QDs. The signal (best measured at excitation/emission peak wavelengths of 365/440 nm) increases linearly in the 0.5 to 10 U·L-1 ALP activity range, with a detection limit of 0.2 U·L-1. The method was applied to the determination of ALP activity in human serum samples and demonstrated satisfactory results. Graphical abstract The fluorescence of chromate-loaded tungsten disulfide quantum dots (QDs) is quenched but restored after reaction with ascorbic acid that is formed by the catalytic action of alkaline phosphatase (ALP) on ascorbic acid 2-phosphate (AAP). The increase in fluorescence can be related to the activity of ALP.

A highly selective colorimetric sulfide assay based on the inhibition of the peroxidase-like activity of copper nanoclusters.

The authors report that sulfide ions are capable of inhibiting the peroxidase-like activity of copper nanoclusters (CuNCs). The catalytic activity of CuNCs toward the oxidation of the chromogenic substrate 3,3',5,5'-tetramethylbenzidine by H2O2 is remarkably decreased in the presence of sulfide. Based on this finding, a colorimetric assay was developed for the rapid determination of sulfide. Best operated at a wavelength of 652 nm, it has a 0.5 µM detection limit. The method is highly selective and has been successfully applied to the quantification of sulfide in environmental water samples. Graphical abstract The catalytic activity of CuNCs toward the oxidation of 3,3',5,5'-tetramethylbenzidine by H2O2 is remarkably decreased in the presence of sulfide ions. This finding has been applied to design a method for colorimetric quantification of sulfide ions in environmental samples.

MoS2 quantum dots modified with a labeled molecular beacon as a ratiometric fluorescent gene probe for FRET based detection and imaging of microRNA.

A dual-channel ratiometric nanoprobe is described for detection and imaging of microRNA. It was prepared from MoS2 quantum dots (QDs; with blue emission and excitation/emission peaks at 310/398 nm) which acts as both the gene carrier and as a donor in fluorescence resonance energy transfer (FRET). Molecular beacons containing loops for molecular recognition of microRNA and labeled with carboxyfluorescein (FAM) were covalently attached to the MoS2 QDs and serve as the FRET acceptor. In the absence of microRNA, the nanoprobe exhibits low FRET efficiency due to the close distance between the FAM tag and the QDs. Hybridization with microRNA enlarges the distance between QD and beacon. This results in an enhancement of the FRET efficiency of the nanoprobe. The ratio of green and blue fluorescence (I520/I398) increases linearly in the 5 to 150 nM microRNA concentration range in both aqueous solution and diluted artificial cerebrospinal fluid. The limit of detection (LOD) is as low as 0.38 nM and 0.52 nM, respectively. Other features of this nanoprobe include (a) excellent resistance to nuclease-induced false positive signals and (b) the option to use it for distinguishing different cell lines by in-situ imaging of intracellular microRNAs. Graphical abstract Schematic of a dual-channel photoluminescence nanoprobe for the determination of microRNA-21 (miR-21) by monitoring the microRNA-triggered enhancement of the fluorescence resonance energy transfer (FRET) efficiency between MoS2 QDs and carboxyfluorescein-labeled molecular beacons.

Water-dispersed fluorescent silicon nanodots as probes for fluorometric determination of picric acid via energy transfer.

Water-dispersed fluorescent silicon nanodots (SiNDs) were synthesized by a one-pot hydrothermal method starting from tetraethyl orthosilicate (TEOS) as silicon source and trisodium citrate as reducing reagent. The method is simple and convenient. The SiNDs, with excitation/emission peaks at 347/440 nm and with fluorescence quantum yield of 18% are shown to be viable fluorescent probes for picric acid (PA). The SiNDs strongly bind PA, and their blue fluorescence is quenched. The distance between the donor and acceptor (R0 value) is calculated from fluorescence data to be 2.1 nm. A fluorometric method was worked out that has a linear response in the 8 nM to 50 µM PA concentration range and a 0.92 nM limit of detection. The method has a fast response (2 min) and is well selective over other nitroaromatic compounds and metal ions. The average recoveries from spiked lake water samples ranged between 98.4 and 100.8%. Graphical abstract Water-dispersed fluorescent silicon nanodots (SiNDs) are synthesized using tetraethyl orthosilicate (TEOS) and trisodium citrate. Based on spectral overlap of fluorescent spectrum of SiNDs and absorption spectrum of picric acid (PA), fluorometric determination of PA at concentrations as low as 0.92 nM is achieved.

Recent development of carbon electrode materials and their bioanalytical and environmental applications.

Carbon materials have been extensively investigated due to their diversity, favorable properties, and active applications including electroanalytical chemistry. This critical review discusses new synthetic methods, novel carbon materials, new properties and electroanalytical applications of carbon materials particularly related to the preparation as well as bioanalytical and environmental applications of highly oriented pyrolytic graphite, graphene, carbon nanotubes, various carbon films (e.g. pyrolyzed carbon films, boron-doped diamond films and diamond-like carbon films) and screen printing carbon electrodes. Future perspectives in the field have also been discussed (366 references).

Topoisomerase-Based Preparation and AFM Imaging of Multi-Interlocked Circular DNA.

Multi-interlocked circular DNA structures have been in high demand for fabricating complicated functional DNA architectures and nanodevices such as molecular switches, shuttles, and motors. Even though various innovative methods have been developed in the past, creation of multi-interlocked circular DNA structures with defined numbers of DNA molecules and linking patterns is still a challenging task nowadays. Here, we propose a top-down decatenation of kinetoplast DNA as a new approach for creating multi-interlocked circular DNA structures. Through optimizing the amount and reaction time of topoisomerase II, we synthesized completely mutually interlocked tricircular, tetra-circular, and oligo-circular DNA structures, which have not yet been acquirable through any other existing synthetic means. The catenation structures of multiple circular DNA were further verified through atomic force microscopic analysis of the backbone overlapping patterns and the circumference. It accordingly is our expectation that the top-down enzymatic approaches could offer a highly interlocked network with defined numbers of circular DNA with simple protocols, and could consequently be beneficial to the design and fabrication of sophisticated functional molecules and nanodevices in the areas of supramolecular chemistry, DNA nanotechnology, and material science.

Switchable catalytic DNA catenanes.

Two-ring interlocked DNA catenanes are synthesized and characterized. The supramolecular catenanes show switchable cyclic catalytic properties. In one system, the catenane structure is switched between a hemin/G-quadruplex catalytic structure and a catalytically inactive state. In the second catenane structure the catenane is switched between a catalytically active Mg(2+)-dependent DNAzyme-containing catenane and an inactive catenane state. In the third system, the interlocked catenane structure is switched between two distinct catalytic structures that include the Mg(2+)- and the Zn(2+)-dependent DNAzymes.

Dual switchable CRET-induced luminescence of CdSe/ZnS quantum dots (QDs) by the hemin/G-quadruplex-bridged aggregation and deaggregation of two-sized QDs.

The hemin/G-quadruplex-catalyzed generation of chemiluminescence through the oxidation of luminol by H2O2 stimulates the chemiluminescence resonance energy transfer (CRET) to CdSe/ZnS quantum dots (QDs), resulting in the luminescence of the QDs. By the cyclic K(+)-ion-induced formation of the hemin/G-quadruplex linked to the QDs, and the separation of the G-quadruplex in the presence of 18-crown-6-ether, the ON-OFF switchable CRET-induced luminescence of the QDs is demonstrated. QDs were modified with nucleic acids consisting of the G-quadruplex subunits sequences and of programmed domains that can be cross-linked through hybridization, using an auxiliary scaffold. In the presence of K(+)-ions, the QDs aggregate through the cooperative stabilization of K(+)-ion-stabilized G-quadruplex bridges and duplex domains between the auxiliary scaffold and the nucleic acids associated with the QDs. In the presence of 18-crown-6-ether, the K(+)-ions are eliminated from the G-quadruplex units, leading to the separation of the aggregated QDs. By the cyclic treatment of the QDs with K(+)-ions/18-crown-6-ether, the reversible aggregation/deaggregation of the QDs is demonstrated. The incorporation of hemin into the K(+)-ion-stabilized G-quadruplex leads to the ON-OFF switchable CRET-stimulated luminescence of the QDs. By the mixing of appropriately modified two-sized QDs, emitting at 540 and 610 nm, the dual ON-OFF activation of the luminescence of the QDs is demonstrated.

"Light-on" sensing of antioxidants using gold nanoclusters.

Depletion of intracellular antioxidants is linked to major cytotoxic events and cellular disorders, such as oxidative stress and multiple sclerosis. In addition to medical diagnosis, determining the concentration of antioxidants in foodstuffs, food preservatives, and cosmetics has proved to be very vital. Gold nanoclusters (Au-NCs) have a core size below 2 nm and contain several metal atoms. They have interesting photophysical properties, are readily functionalized, and are safe to use in various biomedical applications. Herein, a simple and quantitative spectroscopic method based on Au-NCs is developed to detect and image antioxidants such as ascorbic acid. The sensing mechanism is based on the fact that antioxidants can protect the fluorescence of Au-NCs against quenching by highly reactive oxygen species. Our method shows great accuracy when employed to detect the total antioxidant capacity in commercial fruit juice. Moreover, confocal fluorescence microscopy images of HeLa cells show that this approach can be successfully used to image antioxidant levels in living cells. Finally, the potential application of this "light-on" detection method in multiple logic gate fabrication was discussed using the fluorescence intensity of Au-NCs as output.

Boron nanosheets as a phosphatase mimicking nanozyme with ultrahigh catalytic activity for prodrug-based cancer therapy.

Boron nanosheets (BNSs) are reported as a new phosphatase mimicking nanozyme. Surprisingly, the catalytic rate of BNSs is up to 17 times those of known phosphatase mimicking nanozymes. By adding polyols and Lewis bases, the catalytic activity of BNSs was attributed to the Lewis acidity of the B centers of the BNSs. Theoretical investigation shows that the B centers are responsible for the catalytic hydrolysis of phosphoesters. Moreover, the biomimetic activity of the BNSs was further explored for enhancing anticancer therapy through nanozyme-catalyzed prodrug conversion.

Scaling up of a Self-Confined Catalytic Hybridization Circuit for Robust microRNA Imaging.

The precise regulation of cellular behaviors within a confined, crowded intracellular environment is highly amenable in diagnostics and therapeutics. While synthetic circuitry system through a concatenated chemical reaction network has rarely been reported to mimic dynamic self-assembly system. Herein, a catalytic self-defined circuit (CSC) for the hierarchically concatenated assembly of DNA domino nanostructures is engineered. By incorporating pre-sealed symmetrical fragments into the preying hairpin reactants, the CSC system allows the hierarchical DNA self-assembly via a microRNA (miRNA)-powered self-sorting catalytic hybridization reaction. With minimal strand complexity, this self-sustainable CSC system streamlined the circuit component and achieved localization-intensified cascaded signal amplification. Profiting from the self-adaptively concatenated hybridization reaction, a reliable and robust method has been achieved for discriminating carcinoma tissues from the corresponding para-carcinoma tissues. The CSC-sustained self-assembly strategy provides a comprehensive and smart toolbox for organizing various hierarchical DNA nanostructures, which may facilitate more insights for clinical diagnosis and therapeutic assessment.

Cathodic electrochemiluminescence and reversible electrochemistry of [Ru(bpy)3](2+/1+) in aqueous solutions on tricresyl phosphate-based carbon paste electrode with extremely high hydrogen evolution potential.

Many cathodic electrochemiluminescence (ECL) systems require very negative potentials; it is difficult to achieve stable cathodic ECL in aqueous solutions because of hydrogen evolution and instability of intermediates. In this study, tricresyl phosphate-based carbon paste electrode (CPE) was used to achieve cathodic ECL. It exhibits no obvious hydrogen evolution even at a potential up to -1.6 V and dramatically stabilizes electrogenerated [Ru(bpy)3](+). Therefore, a reversible wave of [Ru(bpy)3](2+/1+) in aqueous solutions at carbon electrode has been observed for the first time, and cathodic ECL of [Ru(bpy)3](2+)/S2O8(2-) has been achieved. Under the optimum conditions, the plots of the ECL versus the concentration of S2O8(2-) are linear in the range of 10(-6) to 10(-2) M with the detection limit of 3.98 × 10(-7) M. Common anions have no effect on the ECL intensity of the [Ru(bpy)3](2+)/S2O8(2-) system. Since CPEs have been widely used, CPEs with high hydrogen evolution potential are versatile platforms for electrochemical study and cathodic ECL study.

Ratiometric fluorescent detection of total phosphates in frozen shrimp samples using catalytic active Zr(IV) modified gold nanoclusters.

Phosphate salts are important food additives in a variety of foods. In this study, the Zr(IV) modified gold nanoclusters (Au NCs) were prepared for ratiometric fluorescent sensing of phosphate additives in seafood samples. Compared with bare Au NCs, the synthesized Zr(IV)/Au NCs showed stronger orange fluorescence at 610 nm. On the other hand, the Zr(IV)/Au NCs retained the phosphatase-like activity of Zr(IV) ions and could catalyze the hydrolysis of fluorescent substrate 4-methylumbelliferyl phosphate to produce blue emission at 450 nm. The addition of phosphate salts could effectively inhibit the catalytic activity of Zr(IV)/Au NCs, resulting the fluorescence decrease at 450 nm. However, the fluorescence at 610 nm almost unchanged upon the addition of phosphates. Based on this finding, the ratiometric detection of phosphates using the fluorescence intensity ratio (I450/I610) was demonstrated. The method has been further applied for sensing total phosphates in frozen shrimp samples with satisfactory results.