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Herein, a novel fluorescence nanosensor using intrinsic fluorescent polydopamine nanoparticles (PDA NPs) as an effective signal reporter has been constructed for the simple, rapid and sequential detection of mercury ions (Hg2+) and L-ascorbic acid (AA) based on a coordination effect and redox reaction. The fluorescence of the PDA NPs could be specifically quenched by Hg2+ through intense coordination effects between the Hg2+ and the groups (catechol, amine, ketone and imine) on the surface of the PDA NPs. However, when AA and Hg2+ coexisted in solution, the fluorescence of the PDA NPs pronouncedly recovered via the redox reaction of Hg2+, with it being reduced to Hg0 by AA. The fluorescence quenching mechanism of Hg2+ towards the PDA NPs and the redox reaction between Hg2+ and AA were also fully investigated. The nanosensor exhibited high sensitivity and desirable selectivity for Hg2+ and AA detection. Moreover, the strategy was successfully explored in real samples (tap water, lake water and human serum samples) with satisfactory recoveries. The developed nanosensor provides new sights and good inspiration for Hg2+ and AA detection under real conditions.

Herein, we simply synthesized intrinsic fluorescent polydopamine nanoparticles (PDA NPs) in sodium hydroxide solution (NaOH, pH 11), and constructed a new fluorescence nanoplatform for the detection of alkaline phosphatase (ALP) using PDA NPs as an effective signal reporter. The nanoplatform was constructed by the combination of enzymatic hydrolysis of ALP to the substrate L-ascorbic acid-2-phosphate (AA2P) and the chemical redox reaction between L-ascorbic acid (AA) and mercury ion (Hg2+). The fluorescence of PDA NPs could be effectively quenched by Hg2+ through the coordination effect between Hg2+ and the functional groups on the surface of PDA NPs. However, the quenching effect was greatly inhibited by the addition of AA into the solution. Based on this point, the activity of ALP could be monitored by hydrolysis of the substrate AA2P to AA and the fluorescence output of PDA NPs. The nanoplatform exhibited high sensitivity and desirable selectivity for ALP detection. With a wide linear range of 0 to 18 U L−1, a detection limit of 0.4 U L−1 was achieved using the developed nanosensor. The proposed method could not only be used to screen the inhibitor of ALP but also be used to detect ALP activity in human serum samples successfully. Moreover, the strategy can easily be expanded to determining other kinds of enzymes participating in AA-generation reactions.

A novel fluorescence sensing platform for ultrasensitive detection of S1 nuclease activity has been constructed based on MnO2 nanosheets and FAM labeled single-stranded DNA (FAM-ssDNA). In this system, MnO2 nanosheets were found to have different adsorbent ability toward ssDNA and mono- or oligonucleotide fragments. FAM-ssDNA could adsorb on MnO2 nanosheets and resulted in significant fluorescence quenching through fluorescence resonance energy transfer (FRET), while mono- or oligonucleotide fragments could not adsorb on MnO2 nanosheets and still retained strong fluorescence emission. With the addition of S1 nuclease, FAM-ssDNA was cleaved into mono- or oligonucleotide fragments, which were not able to adsorb on MnO2 nanosheets and the fluorescence signal was never quenched. The different fluorescence intensity allowed for examination of S1 nuclease activity. The developed method can detect S1 nuclease activity in the range of 0 - 20 U mL-1 with a detection limit of 0.05 U mL-1. Benefits of the system include less time-consuming processes and more simple design compared to other endonuclease assays. Satisfactory performance for S1 nuclease in complex samples has been successfully demonstrated with the system. The developed assay could potentially provide a new platform in bioimaging and clinical diagnosis.

Polydopamine (PDA) nanoparticles, as a kind of popular polymer material, have attracted a great deal of attention from various areas including materials science, biomedicine, energy, environmental science and so on owing to their striking physicochemical properties. Herein, we reported for the first time the synthesis of intrinsic fluorescent PDA nanoparticles using MnO2 as an oxidant. In the presence of MnO2, dopamine was quickly oxidized into its quinone derivative, and autopolymerized into fluorescent PDA nanoparticles. Using fluorescent PDA nanoparticles as a fluorescence signal indicator, we further established a cost-effective sensor for rapid, sensitive and selective sensing of reduced glutathione (GSH) based on the redox reaction between MnO2 and GSH, and the key role of MnO2 in the formation of fluorescent PDA nanoparticles. GSH has the capability of reducing MnO2 into Mn(2+), which inhibited the formation of the fluorescent PDA nanoparticles. Thus, the concentration of GSH was directly related to the decreased fluorescence signal intensity of the PDA nanoparticles. The sensor showed good sensing performance for GSH detection with high sensitivity and desirable selectivity over other potential interfering species. Additionally, the sensor exhibited excellent practical applications for GSH detection in human whole blood samples, which presents potential applications in biological detection and clinical diagnosis.

A G-quadruplex-based, label-free fluorescence assay was demonstrated for the detection of adenosine triphosphate (ATP). A double-stranded DNA (dsDNA), hybridized by ATP-aptamer and its complementary sequence, was employed as a substrate for ATP binding. SYBR Green I (SG I) was a fluorescent probe and exonuclease III (Exo III) was a nuclease to digest the dsDNA. Consequently, in the absence of ATP, the dsDNA was inset with SG I and was digested by Exo III, resulting in a low background signal. In the presence of ATP, the aptamer in dsDNA folded into a G-quadruplex structure that resisted the digestion of Exo III. SG I was inserted into the structure, showing high fluorescence. Owing to a decrease of the background noise, a high signal-to-noise ratio could be obtained. This sensor can detect ATP with a concentration ranging from 50 μM to 5 mM, and possesses a capacity for the sensitive determination of other targets.

Blood glucose monitoring has attracted extensive attention because diabetes mellitus is a worldwide public health problem. Here, we reported an upconversion fluorescence detection method based on manganese dioxide (MnO2)-nanosheet-modified upconversion nanoparticles (UCNPs) for rapid, sensitive detection of glucose levels in human serum and whole blood. In this strategy, MnO2 nanosheets on the UCNP surface serve as a quencher. UCNP fluorescence can make a recovery by the addition of H2O2, which can reduce MnO2 to Mn(2+), and the glucose can thus be monitored based on the enzymatic conversion of glucose by glucose oxidase to generate H2O2. Because of the nonautofluorescent assays offered by UCNPs, the developed method has been applied to monitor glucose levels in human serum and whole blood samples with satisfactory results. The proposed approach holds great potential for diabetes mellitus research and clinical diagnosis. Meanwhile, this nanosystem is also generalizable and can be easily expanded to the detection of various H2O2-involved analytes.

A robust fluorescent sensing platform for ultrasensitive assay of nuclease has been established based on rolling circle amplification and exonuclease III-aided recycling amplification. Rolling circle amplification (RCA) is an isothermal DNA amplification process that can convert a short DNA primer into a long single-stranded DNA containing a large amount of tandem repeats. In this work, the substrate DNA (sDNA) of S1 nuclease was designed as the primer to generate RCA products that can hybridize with the pre-quenched TaqMan probes and form recessed 3′ terminus double-stranded DNAs. In the presence of exonuclease III (Exo III), the TaqMan probes were digested from the 3′-hydroxyltermini, releasing the fluorophore and generating enhanced fluorescence signals. Meanwhile, the RCA products with 3′ protruding ends were liberated and hybridized with other TaqMan probes, triggering another cycles and obtaining remarkablely increasing fluorescence. However, in the presence of S1 nuclease, sDNA was cleaved into mono- or short-oligonucleotides pieces, which could not cause the RCA reaction and subsequent Exo III-aided recycling amplification reaction, resulting in extremely weak fluorescence. The fluorescence intensity gradually reduced with increasing concentration of S1 nuclease. Due to the double signal amplification, the developed method was demonstrated to exhibit exceedingly excellent sensitivity with a detection limit of 5 × 10−7 U μL−1. The system was also used to establish a turn-on biosensor of ATP since ATP can prevent the S1 nuclease from digesting the sDNA. ATP can be detected in a range of 5 × 10−4 to 0.2 mM with a detection limit of 5 × 10−4 μM and good selectivity. Moreover, the sensing system was used for the real sample analysis with satisfied results.

Ascorbic acid (AA), a potent antioxidant readily scavenging reactive species, is a crucial micronutrient involved in many biochemical processes. Here, we have developed a cobalt oxyhydroxide (CoOOH)-modified upconversion nanosystem for fluorescence sensing of AA activity in human plasma. The nanosystem consists of upconversion nanoparticles (UCNPs) NaYF4:30% Yb,0.5% Tm@NaYF4, which serve as energy donors, and CoOOH nanoflakes formed on the surface of UCNPs, which act as efficient energy acceptors. The fluorescence resonance energy transfer (FRET) process from the UCNPs to the absorbance of the CoOOH nanoflakes occurs in the nanosystem. The AA-mediated specific redox reaction reduces CoOOH into Co(2+), leading to the inhibition of FRET, and resulting in the recovery of upconversion emission spectra. On the basis of these features, the nanosystem can be used for sensing AA activity with sensitivity and selectivity. Moreover, due to the minimizing background interference provided by UCNPs, the nanosystem has been applied to monitoring AA levels in human plasma sample with satisfactory results. The proposed approach may potentially provide an analytical platform for research and clinical diagnosis of AA related diseases.

Endonuclease plays a vital role in a variety of biological processes and the assay of endonuclease activity and inhibitors is of high importance in the fields ranging from biotechnology to pharmacology. Howerer, traditional techniques usually suffer from time intensive, laborious, and cost-expensive. This work aims to develop a facile and sensitive method for endonuclease activity assay by making use of the fluorescence enhancement effect when DNA–silver nanoclusters (DNA–Ag NCs) are in proximity to guanine-rich DNA sequences. The system mainly consists of block DNA (B-DNA), G-DNA and Ag-DNA. B-DNA serves as the substrate of the endonuclease (S1 nuclease as the model enzyme). G-DNA, which is predesigned entirely complementary to B strand, contains a guanine-rich overhang sequence and hybridization part at the 5′-end. Ag–DNA involves a sequence for Ag NCs synthesis and a sequence complementary to the hybridization part of the G-DNA. In the “off” state, B-DNA plays the role as a blocker that inhibit the proximity between Ag NCs and guanine-rich DNA sequences, resulting in a low fluorescence readout. However, if S1 nuclease is introduced into the system, B-DNA was cleaved into mono- or short-oligonucleotides fragments, which could not hybridize with G-DNA. As a result, the subsequent addition of DNA–Ag NCs could bring guanine-rich DNA sequences close to the Ag NCs, accompanied by a significant fluorescence enhancement. Therefore, endonuclease activity could be successfully quantified by monitoring the variation in fluorescence intensity. In addition, this approach can also be applied for inhibitor screening of endonuclease. This label-free and turn-on fluorescent assays employing the mechanism proposed here for the detection of nuclease and inhibitors turn out to be sensitive, selective, and convenient.

We have developed a reliable, sensitive, label-free and turn-on sensor for H2O2 and glucose based on the cleavage of ssDNA by ˙OH and the fluorescence enhancement effect when guanine-rich (G-rich) DNA sequences are in proximity to DNA–silver nanoclusters (DNA–Ag NCs). In addition, we have also proved that ˙OH is indeed produced in the sensing system by adding antioxidants.

A novel fluorescence sensing platform for ultrasensitive detection of S1 nuclease activity has been constructed based on MnO

Sludge is an important source of greenhouse gas (GHG) emissions, both in the form of direct process emissions and as a result of indirect carbon-derived energy consumption during processing. In this study, the carbon budgets of two sludge disposal processes at two well-known sludge disposal sites in China (for biodrying and heat-drying pretreatments, both followed by mono-incineration) were quantified and compared. Total GHG emissions from heat drying combined with mono-incineration was 0.1731 tCOSludge treatment results in direct and indirect greenhouse gas (GHG) emissions. Moisture reduction followed by incineration is commonly used to dispose of sludge in China; however, few studies have compared the effects of different drying pretreatment options on GHG emissions during such processes. Therefore, in this study, the carbon budgets of sludge incineration were analyzed and compared following different pretreatment drying technologies (biodrying and heat drying). The results indicate that biodrying combined with incineration generated approximately half of the GHG emissions compared to heat drying followed by incineration. Accordingly, biodrying may represent a more environment-friendly sludge pretreatment prior to incineration.

Endonuclease IV (Endo IV), as a DNA repairing enzyme, plays a crucial role in repairing damaged DNA comprising abasic sites to maintain genomic integrity. The cleaving capability of Endo IV to apurinic/apyrimidinic sites (AP) in single-stranded DNA (ssDNA) was demonstrated. It was found that Endo IV has considerably high cleaving activity to AP sites in ssDNA compared with that in double-stranded DNA (dsDNA). The unique feature of Endo IV in cleaving AP sites in ssDNA was further applied to construct a novel dual signal amplified sensing system for highly sensitive enzyme and protein detection by a combination of exonuclease III (Exo III)-aided cyclic amplification reaction and a rolling circle replication (RCR) technique, which showed a good sensing performance with a detection limit of 0.008 U mL(-1) for Endo IV and 2.5 pM for streptavidin. In addition, the developed method had considerably high specificity for Endo IV and streptavidin over other potential interferences. The developed strategy indeed provides a novel platform for protein and enzyme assays and may find a broad spectrum of applications in bioanalysis, disease diagnosis, and drug development.

Blood glucose detecting has aroused considerable attention because diabetes mellitus has become a worldwide publish health problem. Herein, we construct an exceptionally simple upconverting hybrid nanocomposite, composed of DNA-templated Ag nanoparticles (DNA-AgNPs) and NaYF4:Yb/Tm@NaYF4 core-shell upconversion nanoparticles (UCNPs), for the sensing of H2O2 and glucose. In this design, UCNPs with bared surface act as the donor, and DNA-AgNPs serve as efficient quenchers. DNA-AgNPs can be directly assembled on the bared surface of UCNPs, which further decreases the distance of donor-to-acceptor. The formation of DNA-AgNPs/UCNP nanocomposite results in luminescence quenching of UCNP by DNA-AgNPs through luminescence resonance energy transfer (LRET). Upon H2O2 addition, AgNPs can be etched and transformed into Ag(+), leading to inhibition of the LRET process and causing the recovery of upconversion luminescence. Based on the conversion of glucose into H2O2 by glucose oxidase, the DNA-AgNPs/UCNP nanocomposite can also be exploited for glucose sensing. Moreover, due to the non-autofluorescence offered by UCNPs, the approach developed can be applied to monitor glucose levels in human serum samples with satisfactory results.

Human 8-oxoguanine DNA glycosylase (hOGG1) plays a crucial role in maintaining the genomic integrity of living organisms for its capability of repairing DNA oxidative damage. The expression level of hOGG1 is closely associated with many diseases including various kinds of cancers. In this study, a novel "light-up" sensor based on target-induced formation of 5' phosphorylated probe and autocatalytic DNAzyme-generated rolling circle amplification has been developed for highly sensitive human 8-oxoguanine DNA glycosylase (hOGG1) activity assay. The approach reaches detection limit as low as 0.001U/mL for hOGG1 via scarcely increased background signal and dual signal amplification strategy. To the best of our knowledge, it is one of the most sensitive methods for the detection of base excision repair enzyme. Moreover, the approach shows excellent specificity over other nonspecific enzymes would interfere with the assay and holds great promise for application in real sample analysis. Hence, the proposed method provides a highly sensitive, selective, and desirable hOGG1 sensing platform.

Phospholipase D (PLD) is a critical component of intracellular signal transduction and has been implicated in many important biological processes. It has been observed that there are abnormalities in PLD expression in many human cancers, and PLD is thus recognized as a potential diagnostic biomarker as well as a target for drug discovery. We report for the first time a phospholipid-modified nanoprobe for ratiometric upconversion fluorescence (UCF) sensing and bioimaging of PLD activity. The nanoprobe can be synthesized by a facile one-step self-assembly of a phospholipid monolayer composed of poly(ethylene glycol) (PEG)ylated phospholipid and rhodamine B-labeled phospholipid on the surface of upconversion nanoparticles (UCNPs) NaYF4: 20%Yb, 2%Er. The fluorescence resonance energy transfer (FRET) process from the UCF emission at 540 nm of the UCNPs to the absorbance of the rhodamine B occurs in the nanoprobe. The PLD-mediated hydrolysis of the phosphodiester bond makes rhodamine B apart from the UCNP surface, leading to the inhibition of FRET. Using the unaffected UCF emission at 655 nm as an internal standard, the nanoprobe can be used for ratiometric UCF detection of PLD activity with high sensitivity and selectivity. The PLD activity in cell lysates is also determined by the nanoprobe, confirming that PLD activity in a breast cancer cell is at least 7-fold higher than in normal cell. Moreover, the nanoprobe has been successfully applied to monitoring PLD activity in living cells by UCF bioimaging. The results reveal that the nanoprobe provides a simple, sensitive, and robust platform for point-of-care diagnostics and drug screening in biomedical applications.