Aggregation-induced emission for the detection of peptide ligases with improving ligation efficiency.

BACKGROUND: Monitoring peptide ligase activity is of great significance for biological research, medical diagnosis, and drug discovery. The current methods for the detection of peptide ligases suffer from the limitations of high background signal, elaborate design of substrate, and high reversibility of ligation reaction. In this work, we proposed a simple and sensitive method for ligase detection with reducing ligation reversibility on the basis of aggregation-induced emission (AIE) mechanism. RESULTS: The peptide probes labeled with AIE luminogens (AIEgens) were water-soluble and emitted weak fluorescence. After ligation reaction, the enzymatic products with AIEgens showed high hydrophobicity and could readily assembly into aggregates, thus lighting up the fluorescence. More interestingly, the formation of aggregates pushed the equilibrium to the generation of the desired ligation products, thus improving the catalytic efficiency by driving the reaction towards completion. The ligation reaction conversion rate (>80 %) is significantly higher than that without blocking the reversibility with additional treatment. With sortase A (SrtA) as the analyte example, the detection limit of this method was found to be 0.01 nM with a linear range of 0-50 nM. The system was applied to evaluate the inhibition efficiency of berberine chloride and quercetin and determine the activity of SrtA in serum, lysate and Staphylococcus aureus with satisfactory results. SIGNIFICANCE: This study indicated that the ligation efficiency and detection sensitivity can be improved by reducing ligation reversibility through AIE phenomenon. The proposed strategy could be used for the detection of other peptide ligases by adopting sequence-specific peptide substrates.

Reductive dechlorination of trichloroethene by sulfided microscale zero-valent iron in fresh and saline groundwater: Reactivity, pathways, and selectivity.

S/mZVI is a promising material for groundwater remediation due to its excellent properties. However, the reactivity and electron selectivity toward target contaminant are critical. Thus, this study investigated the effect of complex groundwater chemistries (Milli-Q water, fresh groundwater and saline groundwater) on the reactivity of S/mZVI toward trichloroethylene (TCE), dechlorination pathway, hydrogen evolution kinetic, electron efficiency and aging behaviors. Results showed that sulfidation appreciably increased the reactivity and electron selectivity. The major degradation product of TCE dechlorination by S/mZVI was acetylene, which was consistent with TCE dechlorination by ß-elimination. Moreover, reductive ß-elimination was still the dominant dechlorination pathway for the application of S/mZVI in three groundwater conditions. However, the rates and the quantities of major products from TCE degradation varied significantly. S/mZVI in saline groundwater can maintain the reactivity towardTCE due to the protection of Fe0 by Fe3O4 deposited on the surface. Thus, the higher TCE removal efficiency and less hydrogen accumulation resulted in the greatest electron efficiency (4.3-79.2%). Overall, S/mZVI was more effective for the application in saline groundwater. This study proved insight into the comprehensive evaluation and implications for the application of S/mZVI based technologies in saline contaminated groundwater.

Determination of trace bisphenols in milk based on Fe3O4@NH2-MIL-88(Fe)@TpPa magnetic solid-phase extraction coupled with HPLC.

Herein, a covalent organic framework (COF) was grown on a magnetic metal-organic framework (MOF) by a solvothermal method for the efficient extraction of bisphenols (BPs). The magnetic solid-phase extraction (MSPE) of four bisphenols (bisphenol A, bisphenol B, bisphenol AF and bisphenol C) was carried out without adjusting the pH and salt concentration. When 30 mg Fe3O4@NH2-MIL-88(Fe)@TpPa was used to adsorb for 25 min, 6 mL methanol was used to elute for 20 min, and the extract was detected by high-performance liquid chromatography (HPLC). The proposed method has a low detection limit of 0.011-0.036 ng mL-1, a wide linear range of 0.05-100 ng mL-1, and a correlation coefficient (R2) of 0.9980-0.9998. The intra-day and inter-day precisions are 0.74-2.54% and 1.68-3.72%, respectively. Bisphenol A was determined by applying the proposed method to the determination of actual milk samples. The standard addition experiment showed that the relative recovery of the four bisphenols was 85.70-119.7%. Pseudosecond-order, first-order, Langmuir and Freundlich models were applied to explore the adsorption characteristics of Fe3O4@NH2-MIL-88(Fe)@TpPa. In general, the established Fe3O4@NH2-MIL-88(Fe)@TpPa-MSPE-HPLC-UV method exhibits attractive sensitivity, simple manipulation, and excellent reusability, and it has excellent prospects for the detection of trace BPs in complex milk matrices.

High performance isolation of circulating tumor cells by acoustofluidic chip coupled with ultrasonic concentrated energy transducer.

The isolation of circulating tumor cells (CTCs) from whole blood is a challenging task. Although various studies on the separation of CTCs by acoustofluidic devices have been reported, difficulties still persist, such as the complicated equipment, high cost, and difficult operation. Those problems should be resolved urgently. Herein, we developed an acoustofluidic chip separation system coupled with an ultrasonic concentrated energy transducer (UCET) system for efficient separation of CTCs. In the separation system, the acoustically sensitive particles were pre-focused by inertial forces of the PDMS chip channel structure. Then, the particles with different sizes were separated by acoustic radiation forces (ARF). In this study, the circulating tumor cells was simulated (CTCs-like particles) by aminated mesoporous acoustically sensitive particles (MSN@AM) encapsulated carboxylate polystyrene microspheres (PS-COOH). Subsequently, efficient CTCs-like particles separation was achieved by the acoustofluidic chip coupling system. This study effectively separated polystyrene microspheres carrying acoustically sensitive particles (MSN@AM@PS-COOH). However, the MSNs agglomerates and PS microspheres without acoustically sensitive particles did not show phenomenon of separation. This method allows to efficiently separate 2 µm MSNs agglomerates，8.0-8.9 µm PS microspheres and 10-10.5 µm MSN@AM@PS-COOH particles. It is demonstrated that the CTCs-like particles show more sensitive response, longer moving distance, and more obvious separation effect at the condition of the low frequency traveling wave sound field (20 kHz from UCET). This system can maintain the same separation with reduced amount of reagents used for cancer detection. It may provide a reliable basis for sorting out CTCs efficiently from the whole blood of cancer patients.

Fabrication of Two Luminescent Imidazolyl Cadmium-Organic Frameworks and Their Sensing Mechanism for 2,6-Dichloro-4-nitroaniline.

2,6-Dichloro-4-nitroaniline, alias dicloran (DCN), is a broad-spectrum pesticide that can cause irreversible damage to the human body. Therefore, it is of great significance to develop a technology for the rapid and convenient detection of DCN. Luminescent metal organic frameworks have attracted extensive attention in the field of sensing and detection due to their excellent optical properties. In this study, two kinds of 2D Cd-MOFs (CdMOF-1 and CdMOF-2) were developed for the detection of residual DCN in the environment. Both CdMOFs exhibit excellent solvent and acid-base stability and can respond to DCN quickly and sensitively in a short time (30 s). CdMOFs not only have good selectivity and anti-interference toward DCN but also have good reusability. Under the conditions of DCN concentrations of 1-15 and 0.3-30 µM, the change in fluorescence intensity of CdMOF-1 and CdMOF-2 showed a good linear relationship with DCN concentration (R2 = 0.999/0.991), and the detection limits were 0.36 and 0.12 µM, respectively. Through ultraviolet-visible absorption spectroscopy (UV-Vis), X-ray photoelectron spectroscopy, fluorescence lifetime, and density functional theory calculations, it is revealed that the fluorescence quenching mechanisms of DCN for two kinds of Cd-MOFs are competitive absorption and photoinduced electron transfer, and there may be a weak π-π interaction. Finally, it is demonstrated that by using two types of fluorescent CdMOFs to make the fluorescent test paper and detect actual soil, these can be applied to the actual scene and achieve onsite real-time detection.

A novel peptide-based relay fluorescent probe with a large Stokes shift for detection of Hg2+ and S2- in 100 % aqueous medium and living cells: Visual detection via test strips and smartphone.

Herein, a novel relay peptide-based fluorescent probe DGRK was synthesized via solid phase peptide synthesis (SPPS) technology. DGRK exhibited excellent water-solubility, good stability, remarkably large Stokes shift (230 nm) and high selectivity response to Hg2+ with a non-fluorescence complex DGRK-Hg2+ formation via a 1:1 binding mode. Further studies indicated that the DGRK-Hg2+ complex could act as a secondary probe for rapidly and sequentially detecting S2- based on fluorescent "off-on" response, and without interference from a range of anions. The limit of detection (LOD) for Hg2+ and S2- were calculated to be 33.6 nM and 60.9 nM, respectively. In addition, The reversibility of interaction of confirmed that the continuous and reversible recognition behavior of Hg2+ and S2- by the probe DGRK, and could be cycled more than 5 times. In addition, DGRK could be successfully applied to the fluorescence imaging of Hg2+ and S2- in two living cells based on excellent cells permeability and low cytotoxicity. Meanwhile, DGRK was successfully used to create the low-cost and portable test strips for visual detection and rapid analysis under 365 nm UV lamp, and the test strips combined with a smartphone (RGB color) was successfully applied to the semi-quantitative analysis and monitoring of dynamic changes of Hg2+ levels.

A gold-silver bimetallic nanocluster-based fluorescent probe for cysteine detection in milk and apple.

Noble metal nanoclusters have attracted much attention due to their excellent optical properties. In the present work, a silver-doped gold-based bimetallic nanoclusters (Au/Ag NCs) were reasonably designed and prepared through a one-pot method by using 5-mercapto-1-tetrazolea-acetic acid sodium salt (MTAS) as a protector and capping agent. In comparison with the monometallic nanoclusters, Ag-doped metallic nanoclusters show better performance. The particle size of the MTAS-Au/Ag NCs is slightly larger than that of the undoped Au NCs by about 1.86 ± 0.5 nm, and the MTAS-Au/Ag NCs exhibit an emission peak at 635 nm with a quantum yield (QY) of 3.05%. The presence of cysteine (Cys) induces efficient quenching of the photoluminescence of the obtained Au/Ag NCs, achieving the sensitive detection of Cys with a detection limit of 16 nM. The fluorescence quenching rate of the nano fluorescent probe has a linear relationship with the cysteine concentration. Under the best detection conditions, the linear range for Cys detection with MTAS-Au/Ag NCs as a probe is 0.05-25.0 µM. Moreover, this probe has been successfully applied to the analysis of Cys in milk and apples, and a satisfactory recovery rate has been obtained, indicating the effectiveness and reliability of the sensor system for the detection of actual samples.

Peptide-based colorimetric and fluorescent dual-functional probe for sequential detection of copper(â ¡) and cyanide ions and its application in real water samples, test strips and living cells.

A novel dual-functional peptide probe FLH based on fluorescent "on-off-on" strategy and colorimetric visualization method was designed and synthesized. This new probe exhibited highly selective and rapid detection of Cu2+ with significant fluorescent "turn-off" response, with a visible colorimetric change from yellow to orange. The combination ratio of FLH to Cu2+ (1:1) was determined using ESI-HRMS spectra and Job's plot. The fluorescent emission showed a good linear response (R2 = 0.9986) with a low detection limit of 1.5 nM. In addition, the FLH-Cu2+ complex displayed colorimetric changes and a fluorescent "off-on" response toward CN- over a wide pH range from 7 to 12. This detection behavior was observed within 20 s, with a limit of detection (LOD) for CN- at 12.7 nM. Based on stability and accuracy, FLH was next developed as dual-functional test strips, and was also successfully applied to detect Cu2+ and CN- in two actual water samples. More importantly, the cytotoxicity studies indicated that FLH had good biocompatibility and low toxicity, and was successfully utilized for monitoring Cu2+ and CN- in living cells through fluorescence imaging.

A bifunctional peptide-based fluorescent probe for ratiometric and "turn-on" detection of Zn(II) ions and its application in living cells.

In this work, a bifunctional peptide-based fluorescent probe L containing a tetrapeptide scaffold (Pro-Gly-His-Trp-NH2) and a dansyl group was synthesized using solid phase peptide synthesis (SPPS) technology. As designed, L, based on a FRET mechanism, exhibited high selectivity, excellent ratiometric signals, and fast response to Zn2+ in aqueous solutions at an excitation wavelength of 280 nm. In addition, when excited at 320 nm, L exhibited a fluorescent "turn-on" response towards Zn2+ based on PET mechanism. More importantly, the stoichiometry of L and Zn2+ was determined to be 2:1 by fluorescent titration, Job's plot method, and ESI-MS spectrometry. The association constant for Zn2+ ions was determined to be 6.26 × 108 M-2, while the limit of detection (LOD) of L was estimated as 5.43 nM, which is a much lower value than WHO and EPA guidelines for drinking water. Moreover, L was successfully applied to detect both Zn2+ and Cu2+ in living cells due to good biocompatibility and excellent low toxicity.

Recyclable Luminescence Sensor for Dinotefuran in Water by Stable Cadmium-Organic Framework.

Due to the widespread use of dinotefuran around the world, its impact on food and environmental safety has aroused great concern, and the establishment of a rapid and convenient approach for dinotefuran detection is necessary but challenging. Herein, we synthesized a unique three-dimensional framework {[(CH3)2NH2]2[Cd3(BCP)2]·10H2O·3.5DMF}n (1). Single-crystal X-ray analysis indicates that 1 possesses a 4,8-connected anion framework that corresponds to alb topology, with a one-dimensional rectangular channel along the c-axis with the size of 4 Å × 10 Å. Compound 1 displays satisfactory solvent and thermal stability. Luminescent investigations reveal that 1 can selectively detect dinotefuran by fluorescence quenching among other pesticides, displaying excellent anti-interference performance with common ions in water. Importantly, the limit of detection is as low as 2.09 ppm, which is far below the residual concentration of the U.S. food standard. A fluorescence quenching mechanism study shows that there exists competitive energy absorption and static quenching processes. To our knowledge, 1 is the first MOF-based fluorescence probe for dinotefuran detection.

Remediation of trichloroethylene by microscale zero-valent iron aged under various groundwater conditions: Removal mechanism and physicochemical transformation.

Microscale zero-valent iron (mZVI) has been widely used for the in-situ groundwater remediation of various pollutants. However, the aging behavior of injected mZVI particles limits the widespread application in groundwater remediation projects. To assess the long-term reactivity of mZVI particles, the mechanism of trichloroethylene (TCE) degradation by various aged mZVI particles (A-mZVI) was determined by quantitatively evaluating the contributions of chemical reduction and adsorption. Further, this study investigated the physicochemical transformation of mZVI particles aged under various hydraulic conditions (static and dynamic), redox conditions (anoxic and aerobic) and aging durations (152 d and 455 d). The results show that the removal of TCE by different A-mZVI particles increased the sorption capacity in the initial period (0-6 h). However, in the long term, a significant inhibition of TCE removal was observed because of the decreased TCE reduction capacity caused by the hindrance of electron transfer, which was generated by corrosion precipitates. Furthermore, the characterization results demonstrated that despite the significant differences in the apparent morphology of the A-mZVI particles in various groundwater conditions, the final crystal corrosion products were mainly Fe3O4. Thus, the aging and inactivation of mZVI particles on TCE removal were promoted under the aerobic conditions. In addition, the structure of mZVI particles collapsed from the micro- to nanoscale under anaerobic dynamic over 455 d. No substantial impact on the final TCE removal was observed for the A-mZVI particles prepared under various hydraulic conditions and aging times. These findings provide insights regarding the impact mechanisms of corrosion precipitates on the removal of target contaminant and provide implications for long-term mZVI application under various target aquifer conditions.

A dual-functional colorimetric and fluorescent peptide-based probe for sequential detection of Cu2+ and S2- in 100% aqueous buffered solutions and living cells.

Highly sensitive and selectivite detection of copper ions (Cu2+) and hydrogen sulfide (H2S) have become important research topics due to the potential harmful impacts of these chemicals to human health and the environment. In this study, we report the synthesis of a dual-functional peptide-based probe L (FITC-AhxSerSerHis), designed to mimic a copper-sulfur metalloprotein, and capable of continuous detection of Cu2+ and S2- based on colorimetric and fluorescent methods. The new probe L displayed excellent "turn off" fluorescence response and good selectivity for Cu2+ ions via a modification of the tripeptide and fluorescein isothiocyanate group, and produced an obvious color change visible to the naked eye. Furthermore, as an excitable probe, the L-Cu complex could continuously detect S2- with high selectivity and sensitivity in 100% aqueous buffered solutions. The detection limits for fluorescence titration measurements, calculated using the equation 3σ/k, were 76.7 nM (Cu2+) and 27.2 nM (S2-), which were well below U.S. EPA safety levels. In addition, L could be cycled to alternately detect Cu2+ and S2-, thereby making it a promising reversible probe. Moreover, L was successfully applied to monitoring Cu2+ and S2- in live RKO cells through fluorescence imaging, exhibiting low cytotoxicity and good cell permeability.

Highly selective and sensitive detection of hydrogen sulfide in aqueous medium and live cells using peptide-based bioprobe to mimic the binding sites of the ceruloplasmin for Cu(II) ions.

Hydrogen sulfide (H2S) plays a vital role in biological systems, so it is important to develop new methods for monitoring H2S with high selectively and sensitivity. In this study, we report the efficient solid phase synthesis of a new peptide-based bioprobe L that mimics the Cu(II) binding sites of ceruloplasmin. L exhibits highly sensitive colorimetric and fluorescent "turn off" responses to Cu2+ with high selectively among various metal ions over a broad pH range in 100% aqueous solutions. Interestingly, as a new promising analytical bioprobe, the L-Cu ensemble can monitor H2S by both colorimetric and fluorescent changes, and the limit of detection (LOD) is found to be as low as 14.7 nM. The reversibility experiment indicated that L could be recycled many times as reversible bioprobe. It is worth noting that L could be used to detect Cu2+ and H2S specifically in two real samples. Moreover, L exhibited excellent water-solubility and low biotoxicity even at high concentrations based on MTT study, and cell imaging results indicated that it can be applied to detecting Cu(II) and H2S in living RKO cells monitored by confocal fluorescence microscopy imaging.

Enhanced light-emitting diode induced fluorescence detection system with capillary electrophoresis.

An enhanced fluorescence detection system of capillary electrophoresis (CE) was equipped with a concave silver mirror, by which the detection sensitivity of light-emitting diode induced fluorescence (LEDIF) can be increased greatly. The silver concave mirror and the cathode window in photomultiplier tube (PMT) were accurately set face to face at the same axis. When the two labeled tumor markers exactly moved to the center of detection window, the emission from analytes are excitated by LED source. Currently, the analytes may be regarded as a luminescent source point. When the source point exactly moves to the focus of the concave mirror, the emission of the labeled sample was collected effectively, enhanced by convergence and reflected by the concave mirror. Then it was sensitively detected by the PMT. The optical mechanism of enhancing detection sensitivity was explored. A simple comparative test on sensitivity was carried out, which aimed to compare sensitivity of the new detection system with concave mirror to that without concave mirror but the other conditions were kept the same. Two tumor markers labeled with FITC were selected for the test, using the simple LEDIF detect system. The results (LOD, 150 nM for L-Leu and L-Val) showed that the detection sensitivity matched with concave mirror reached more 16 times than the detection method without concave mirror.

Synthesis of N-acetyl-l-cysteine capped Mn:doped CdS quantum dots for quantitative detection of copper ions.

In this work, a new assembled copper ions sensor based on the Mn metal-enhanced fluorescence of N-acetyl-l-cysteine protected CdS quantum dots (NAC-Mn:CdS QDs) was developed. The NAC and Mn:CdS QDs nanoparticles were assembled into NAC-Mn:CdS QDs complexes through the formation of CdS and MnS bonds. As compared to NAC capped CdS QDs, higher fluorescence quantum yields of NAC-Mn:CdS QDs was observed, which is attributed to the surface plasmon resonance of Mn metal. In addition, the fluorescence intensity of as-formed complexes weakened in the presence of copper ions. The decrease in fluorescence intensity presented a linear relationship with copper ions concentration in the range from 0.16-3.36µM with a detection limit of 0.041µM . The characterization of as-formed QDs was analyzed by photoluminescence (PL), ultra violet-visible (UV-vis), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and energy dispersive spectroscopy (EDS) respectively. Furthermore, the recoveries and relative standard deviations of Cu2+ spiked in real water samples for the intra-day and inter-day analyses were 88.20-117.90, 95.20-109.90, 0.80-5.80 and 1.20-3.20%, respectively. Such a metal-enhanced QDs fluorescence system may have promising application in chemical and biological sensors.

Single fiber-in-capillary annular column for gas chromatographic separation.

A method for preparation of a stationary phase-adjustable column with in-column stationary phase-coated fused-silica fiber annular column was successfully developed. The surface of a 0.12 mm o.d. bare optical fiber was first coated with a stationary phase and then inserted into a fused-silica capillary (non-coated or coated) as an annular column for gas chromatographic study. The optical fiber and capillary were coated with polydimethylsiloxane (SE-30) and polyethylene glycol 20 M (PEG-20 M) as nonpolar and polar stationary phases, respectively. Among the investigated annular and open tubular columns, the PEG-20 M-coated fiber-in-PEG-20 M-coated capillary annular column showed the highest column efficiency with a minimum plate height of 0.35 mm and an optimum gas velocity of 25 cm/s. When a SE-30/PEG-20 M-coated fiber-in-uncoated capillary annular column was applied to separate a 9-component complex mixture, the total analysis time was 5.3 min and the column length was 12 m. By contrast, when a SE-30-coated fiber-in-PEG-20 M-coated capillary annular column was used to separate the same 9-component mixture, the analysis time was reduced to 3.5 min and the column length was shortened by half to 6m. Our results show that the stationary phase-coated fiber-in-stationary phase-coated capillary annular column is a better choice for gas chromatographic separation as it is more efficient and flexible. In addition, the proposed annular column design provides flexibility in using two or even more types of stationary phases to achieve optimal analytical separation.