Ultra-sensitive fluorescent biosensor for multiple bacteria detection based on CDs/QDs@ZIF-8 and microfluidic fluidized bed.

An ultra-sensitive fluorescent biosensor based on CDs/QDs@ZIF-8 and microfluidic fluidized bed was developed for rapid and ultra-sensitive detection of multiple target bacteria. The zeolitic imidazolate frameworks (ZIF-8) act as the carrier to encapsulate three kinds of fluorescence signal molecules from the CDs/QDs@ZIF-8 signal amplification system. Besides, three kinds of target pathogenic bacteria were automatically, continuously, and circularly captured by the magnetic nanoparticles (MNPs) in the microfluidic fluidized bed. The neutral Na2EDTA solution was the first time reported to not only dissolve the ZIF-8 frameworks from the MNPs-bacteria-CDs/QDs@ZIF-8 sandwich complexes, but also release the CDs/QDs from sandwich complexes with no loss of fluorescence signal. Due to the advantages of signal amplification and automated sample pretreatment, the proposed fluorescent biosensor can simultaneously detect Escherichia coli O157:H7, Salmonella paratyphi A, and Salmonella paratyphi B as low as 101 CFU/mL within 1.5 h, respectively. The mean recovery in spiked milk samples can reach 99.18%, verifying the applicability of this biosensor in detecting multiple bacteria in real samples.

Chemical characterisation of essential oil from Sambucus williamsii Hance leaves and its hepatoprotective effects.

This study is the first to examine the effect of leaves of Sambucus williamsii Hance essential oil on acute liver injury. According to gas chromatography-mass spectrometry analysis, the major constituents of S. williamsii essential oil (SEO)were (S)-falcarinol (62.66%), 17-pentatriacontene (7.78%) and tetrapentacontane (8.64%). Mice were pre-treated with SEO for 6 days followed by inducing liver injury with CCl4. The results indicated that SEO protected the liver against CCl4-induced injuries. Elevated levels of alanine-aminotransferase (ALT), aspartate amino-transferase (AST), alkaline phosphatase (ALP) in serum were significantly reduced on SEO pre-treatment. SEO pre-treatment significantly inhibited the oxidative stress and inflammation. Furthermore, toll-like receptor-4 (TLR4)/nuclear factor kappa-B (NF-κB) signalling pathways were significantly modulated by SEO in the liver tissue. The findings demonstrate that the essential oil of S. williamsii has enhancing the resistance to CCl4-induced liver injury.

Tuning Spin-Polarized Lifetime at High Carrier Density through Deformation Potential in Dion-Jacobson-Phase Perovskites.

The control of spin relaxation mechanisms is of great importance for spintronics applications as well as for fundamental studies. Layered metal-halide perovskites represent an emerging class of semiconductors with rich optical spin physics, showing potential for spintronic applications. However, a major hurdle arises in layered metal-halide perovskites with strong spin-orbit coupling, where the spin lifetime becomes extremely short due to D'yakonov-Perel' scattering and Bir-Aronov-Pikus at high carrier density. Using the circularly polarized pump-probe transient reflection technique, we experimentally reveal the important scattering for spin relaxation beyond the electron-hole exchange strength in the Dion-Jacobson (DJ)-type 2D perovskites (3AMP)(MA)n-1PbnI3n+1 [3AMP = 3-(aminomethyl)piperidinium, n = 1-4]. Despite a more than 10-fold increase in carrier concentration, the spin lifetimes for n = 3 and 4 are effectively maintained. We reveal neutral impurity and polar optical phonon scatterings as significant contributors to the momentum relaxation rate. Furthermore, we show that more octahedral distortions induce a larger deformation potential which is reflected on the acoustic phonon properties. Coherent acoustic phonon analysis indicates that the polaronic effect is crucial in achieving control over the scattering mechanism and ensuring spin lifetime protection, highlighting the potential of DJ-phase perovskites for spintronic applications.

Ultrafast charge transfer in metal-free H2O2 photoproduction by anhydride modified g-C3N4.

As a low-cost, low toxicity and metal-free catalyst with strong light absorption, graphitic carbon nitride (g-C3N4)-based materials have gained wide attention for efficient H2O2 photocatalysis. However, further investigation regarding the charge transfer process and reaction mechanism of H2O2 photoproduction remains to be completed. In this work, bicyclo[2.2.2]oct-7-ene-2,3,5,6-tetracarboxylic dianhydride (BTDA) modified g-C3N4 is synthesized through a facile one-step dehydration process, and the H2O2 photoproduction could reach 22.5 µmol within 8 hours. The proposed structure of g-BTDA is confirmed by FTIR, XPS and SEM studies. The transient absorption reveals a 20.88 ps charge transfer process caused by the electron withdrawing ability of the CO group, and a 2-electron oxygen reduction pathway is proposed. Our work represents a new strategy for efficient H2O2 photoproduction using easily acquired materials with future application potential.

An immobilization-free electrochemical biosensor based on CRISPR/Cas13a and FAM-RNA-MB for simultaneous detection of multiple pathogens.

To better respond to biosecurity issues, we need to build good technology and material reserves for pathogenic microorganism screening. Here, we designed an electrochemical/optical signal probe with a common fluorophore and an electrochemically active group, breaking the previous perception that the signal probe is composed of a fluorophore and a quenching group and realizing the response of three signals: electrochemistry, fluorescence, and direct observation. Then, we proposed a homogeneous electrochemical nucleic acid detection system based on CRISPR/Cas named "HELEN-CR" by integrating free electrochemical/optical signal probes and Cas13a cleavage, achieving a limit of detection of 1 pM within 25 min. To improve the detection sensitivity, we applied recombinase polymerase amplification to amplify the target nucleic acid, achieving a limit of detection of 30 zM within 45 min. Complemented by our self-developed multi-chamber microfluidic chip and portable electrochemical instrument, simultaneous detection of multiple pathogens can be achieved within 50 min, facilitating minimally trained personnel to obtain detection results quickly in a difficult environment. This study proposes a simple, scalable, and general idea and solution for the rapid detection of pathogenic microorganisms and biosecurity monitoring.

Ultrafast Hole Transfer in Graphitic Carbon Nitride Imide Enabling Efficient H2O2 Photoproduction.

Solar-driven photocatalysis is a promising approach for renewable energy application. H2O2 photocatalysis by metal-free graphitic carbon nitride has been gaining attention. Compared with traditional thermal catalysis, metal-free graphitic carbon nitride photocatalysis could lower material cost and achieve greener production of H2O2. Also, to better guide photocatalyst design, a fundamental understanding of the reaction mechanism is needed. Here, we develop a series of model cost-effective metal-free H2O2 photocatalysts made from graphitic carbon nitride (melem) and common imide groups. With 4,4'-oxydiphthalic anhydride (ODPA)-modified g-C3N4, a H2O2 yield rate of 10781 µmol/h·g·L could be achieved. Transient absorption and ex situ Fourier transform infrared (FTIR) measurements revealed an ultrafast charge transfer from the melem core to water with ∼3 ps to form unique N-OH intermediates. The electron withdrawing ability of the anhydride group plays a role in governing the rate of electron transfer, ensuring efficient charge separation. Our strategy represents a new way to achieve a low material cost, simple synthesizing strategy, good environment impact, and high H2O2 production for renewable energy application.

Nanomaterial catalysts for organic photoredox catalysis-mechanistic perspective.

Solar energy conversions play a vital role in the renewable energy industry. In recent years, photoredox organic transformations have been explored as an alternative way to use solar energy. Catalysts for such photocatalytic systems have evolved from homogeneous metal complexes to heterogeneous nanomaterials over the past few decades. Herein, three important carrier transfer mechanisms are presented, including charge transfer, energy transfer and hot carrier transfer. Several models established by researchers to understand the catalytic reaction mechanisms are also illustrated, which promote the reaction system design based on theoretical studies. New strategies are introduced in order to enhance catalytic efficiency for future prospects.

Wide-Range, Rapid, and Specific Identification of Pathogenic Bacteria by Surface-Enhanced Raman Spectroscopy.

Sensitive, selective, rapid, and label-free detection of pathogenic bacteria with high generality is of great importance for clinical diagnosis, biosecurity, and public health. However, most traditional approaches, such as microbial cultures, are time-consuming and laborious. To circumvent these problems, surface-enhanced Raman spectroscopy (SERS) appears to be a powerful technique to characterize bacteria at the single-cell level. Here, by SERS, we report a strategy for the rapid and specific detection of 22 strains of common pathogenic bacteria. A novel and high-quality silver nanorod SERS substrate, prepared by the facile interface self-assembly method, was utilized to acquire the chemical fingerprint information of pathogens with improved sensitivity. We also applied the mathematical analysis methods, such as the t-test and receiver operating characteristic method, to determine the Raman features of these 22 strains and demonstrate the clear identification of most bacteria (20 strains) from the rest and also the reliability of this SERS sensor. This rapid and specific strategy for wide-range bacterial detection offers significant advantages over existing approaches and sets the base for automated and onsite detection of pathogenic bacteria in a complex real-life situation.

Multicolor Coding Up-Conversion Nanoplatform for Rapid Screening of Multiple Foodborne Pathogens.

Technologies for rapid screening of multiple foodborne pathogens have been urgently needed because of the complex food matrix and high outbreaks of foodborne diseases. In this study, multicolor coding up-conversion nanoparticles (UCNPs) were synthesized and applied for rapid and simultaneous detection of five kinds of foodborne pathogens. The multicolor coding UCNPs were obtained through doping different concentrations of a sensitizer (Yb3+) on the shell of the synthesized NaYF4:Yb3+, Tm3+ (20%/2%)@NaYF4:Yb3+, and Er3+ (x %/2%) core/shell nanocrystals. All the UCNPs could emit red and green luminescence simultaneously once excited with near-infrared wavelength (980 nm), and the ratio of red and green (R/G ratio) emission light intensity of each kind of UCNPs varied depending on the Yb3+ doping concentration. In addition, the magnetic nanoparticles (MNPs) modified with the monoclonal antibodies (mAbs) against the target bacteria were used to capture and separate the bacteria, resulting in obtaining the MNP-bacterium complexes. Different UCNPs with multicolor coding acted as signal probes were also modified with the mAbs to react with the MNP-bacterium complexes to form the MNP-bacterium-UCNP sandwich complexes. After the sandwich complexes were excited with a wavelength of 980 nm, the obtained R/G ratios and the green photoluminescence intensity (PL intensity) could be used to distinguish and quantitatively detect foodborne pathogens, respectively. This proposed nanoplatform could detect five foodborne pathogens simultaneously within 2 h with good sensitivity and specificity, showing great potential for multiplex detection of other targets in the fields of medical diagnosis and food security.

Rapid and quantitative detection of Shiga toxin1 and Shiga toxin2 based on multiple targets UPT-LF assay.

Shiga toxin-producing Escherichia coli (STEC) infection causes a series of diseases that are highly pathogenic and deadly in humans and animals, seriously endangering public health. Of the pathogenic factors within STEC, the two groups of Shiga toxin (Stx) consisting Stx1 and Stx2 plays a prominent role in the pathogenesis of STEC infection. In this study, we developed single-target up-converting phosphor technology-based lateral flow assay (Stx-UPT-LFA) for the rapid detection of Stx1 and Stx2, respectively, and also developed a dual-target Stx1/2-UPT-LFA based on single-target strips to detect of Stx1 and Stx2 at the meantime within 20 min. We choose the purified Stx1 and Stx2 standard samples, and the optimum monoclonal antibody (namely 8E7-E6, 2F6-F8 for Stx1 and S1D8, S2C4 for Stx2) were selected for use in Stx-UPT-LFA in double-antibody-sandwich mode. The sensitivities of single-target Stx-UPT-LFA for both Stx1 and Stx2 were 1 ng mL-1 with accurate quantitation ranges of 1-1000 ng mL-1 and 1-800 ng mL-1 respectively. No false-negative result was found in the Stx2-UPT-LFA even with a high-test concentration up to 1000 ng mL-1. Meanwhile, both targets detection sensitivities for dual-target Stx1/2-UPT-LFA were 5 ng mL-1, and accurate quantitation ranges were 5-1000 ng mL-1 and 5-800 ng mL-1 for standard Stx1 and Stx2 solutions without cross-interference between two targets. Both techniques showed good linearities, with a linear fitting coefficient of determination(r) of 0.9058-0.9918. Therefore, the UPT-LFA could realize simultaneous detection for multiple targets on a single strip and thus to quickly determine the type of infectious Stxs. In addition, the single-target Stx1-UPT-LFA and Stx2-UPT-LFA showed excellent specificity to six toxins, even at high concentrations of 1000 ng mL-1. In conclusion, the developed Stx-UPT-LFA allows the rapid, quantitative, reliable and simultaneous detection of Stx1 and Stx2 within 20 min, providing an alternative method for clinical diagnosis of STEC infection.

Identification of eight pathogenic microorganisms by single concentration-dependent multicolor carbon dots.

The identification of pathogenic microorganisms is crucial to human health and industrial development, however, traditional culture-based methods are laborious and time consuming. We developed a novel method for the identification of pathogenic microorganisms based on single concentration-dependent carbon dots (CDs). The CDs were synthesized using a microwave heating approach without any complicated purification processes and the PL emission wavelength of the CDs could be tuned only by changing the concentration of the solution. The concentration-dependent CDs were applied for multicolor bioimaging and identification of eight kinds of microorganisms within minutes using different fluorescence spectra. This work provides a new approach for the detection of multiplex targets in real samples and lays a foundation for food safety monitoring.

Rapid detection of severe fever with thrombocytopenia syndrome virus (SFTSV) total antibodies by up-converting phosphor technology-based lateral-flow assay.

In this study, an up-converting phosphor technology-based lateral-flow (UPT-LF) assay was developed to detect severe fever with thrombocytopenia syndrome virus (SFTSV) total antibodies rapidly and specifically. SFTSV recombinant N protein (SFTSV-rNP) was coated on analytical membrane for sample capture, up-converting phosphor (UCP) particles were used as the reporter, the luminescence emitted by UCP particles was converted to a measurable signal by a biosensor. The performance of UPT-LF assay was evaluated by testing 302 field serum samples by ELISA (enzyme-linked immunosorbent assay), Western blotting and UPT-LF assay. UPT-LF assay exhibited a lower detection limit than ELISA, and a satisfied level of agreement was exhibited by Kappa statistics (Kappa coefficient = 0.938). Considering Western blotting as the reference for comparison, the sensitivity and specificity of UPT-LF assay could reach 98.31% and 100%. UPT-LF assay showed no specific reaction with hantavirus total serum antibodies, which avoids the misdiagnosis of SFTSV from hantavirus that could cause similar clinical symptoms. UPT-LF assay was able to achieve acceptable results within 15 min and needed only 10 µL sample for each test. As a whole, UPT-LF assay is a candidate method for on-site surveillance of SFTSV total antibodies owing to its excellent sensitivity, specificity, stability, easy operation and for being less time consuming.

An up-converting phosphor technology-based lateral flow assay for point-of-collection detection of morphine and methamphetamine in saliva.

Morphine (Mop) and methamphetamine (Met) are highly addictive drugs worldwide. Point-of-collection testing (POCT) for drug-of-abuse screening is important in abuse/rehabilitation clinics and law-enforcement agencies. We established an up-converting phosphor technology-based lateral flow assay (UPT-LFA) as a point-of-collection testing (POCT) method, namely Mop-UPT-LFA and Met-UPT-LFA, for the detection of morphine and methamphetamine without complicated sample pre-treatment, respectively, in saliva. The sensitivities of the Mop-UPT-LFA and the Met-UPT-LFA were 5 and 10 ng mL-1 with accurate quantitation of 5-100 ng mL-1 and 10-250 ng mL-1 for morphine and methamphetamine, respectively, for a detection time of 15 min. In reference to the detection limits of 20 and 25 ng mL-1 for morphine and methamphetamine, respectively, in the Driving Under the Influence of Drugs, Alcohol and Medicines (DRUID) program of the European Union, the percentage test/control (T/C) ratio of the UPT-LFA between 2 and 15 min reached 101% and 86%, and the UPT-LFA produced accurate qualitative results in 2 min for 100 simulated-saliva samples with the exception of a few weakly positive samples. The sample and sample treating buffer were mixed and added to the test strip, and the test was conducted 15 min later. Although we found no significant difference between the UPT-LFA quantitative test and the liquid chromatography tandem mass spectrometry (LC-MS) test, compared with the latter, the UPT-LFA was substantially faster and had higher detection efficiency. The UPT-LFA showed more accurate qualitative results than the LC-MS for 50 simulated-saliva samples. The ease of operation, high sensitivity, and accuracy of the UPT-LFA make it a valid candidate POCT method for drug-of-abuse screening.

Ultrathin polydopamine film coated gold nanoparticles: a sensitive, uniform, and stable SHINERS substrate for detection of benzotriazole.

In shell-isolated nanoparticle (NP)-enhanced Raman spectroscopy (SHINERS), traditional metal oxide-based shells have inferior chemical inertness, they require strict preparation conditions, and lack specific groups, which lead to their poor selectivity toward target molecules. In this study, ultrathin and compact gold (Au)@polydopamine (PDA) SHINERS NPs were successfully fabricated by a simple self-polymerization technique. High wrapping tendency of PDA, a multifunctional biopolymer, favored the fabrication process. Au@PDA NPs exhibited a typical shell-isolated effect, i.e., Au@PDA NPs with a thick shell (more than 2.3 nm) showed a lower SERS activity, while those with an ultrathin (1.3 nm) shell exhibited higher SERS activity compared to uncoated Au NPs. The Au@PDA SHINERS substrate shows high performance in terms of sensitivity, uniformity, and stability. The relative standard deviations (RSDs) of SERS intensities from ten positions on the same substrate were less than 4%. Their Raman intensities dropped by only 15% over two months. More importantly, the Au@PDA (1.3 nm) SHINERS substrate exhibited high SERS activity for label-free and quantitative detection of benzotriazole (BTA), an important corrosion inhibitor, through utilizing a presumed π-π stacking interaction. A broad linear range from 10-4 to 10-8 M was achieved with a low detection limit (LOD) of 1 nM (0.119 µg L-1). The LOD was not only significantly lower than the maximum allowable level (20 µg L-1) of the Australian government water guide, but also lower than that of some modern methods such as fluorescence, liquid chromatography, and gas chromatography coupled with mass spectrometry. Furthermore, the substrate showed excellent discrimination against other compounds with a single aromatic ring. It is expected that the Au@PDA SHINERS substrate will offer great potential for analysis application in a complicated environmental system.

Purification and characterization of cold-adapted beta-agarase from an Antarctic psychrophilic strain.

An extracellular ß-agarase was purified from Pseudoalteromonas sp. NJ21, a Psychrophilic agar-degrading bacterium isolated from Antarctic Prydz Bay sediments. The purified agarase (Aga21) revealed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 80 kDa. The optimum pH and temperature of the agarase were 8.0 and 30 °C, respectively. However, it maintained as much as 85% of the maximum activities at 10 °C. Significant activation of the agarase was observed in the presence of Mg(2+), Mn(2+), K(+); Ca(2+), Na(+), Ba(2+), Zn(2+), Cu(2+), Co(2+), Fe(2+), Sr(2+) and EDTA inhibited the enzyme activity. The enzymatic hydrolyzed product of agar was characterized as neoagarobiose. Furthermore, this work is the first evidence of cold-adapted agarase in Antarctic psychrophilic bacteria and these results indicate the potential for the Antarctic agarase as a catalyst in medicine, food and cosmetic industries.

Purification and characterization of cold-adapted beta-agarase from an Antarctic psychrophilic strain

An extracellular β-agarase was purified from Pseudoalteromonas sp. NJ21, a Psychrophilic agar-degrading bacterium isolated from Antarctic Prydz Bay sediments. The purified agarase (Aga21) revealed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 80 kDa. The optimum pH and temperature of the agarase were 8.0 and 30 °C, respectively. However, it maintained as much as 85% of the maximum activities at 10 °C. Significant activation of the agarase was observed in the presence of Mg²⁺, Mn²⁺, K⁺; Ca²⁺, Na⁺, Ba²⁺, Zn²⁺, Cu²⁺, Co²⁺, Fe²⁺, Sr²⁺ and EDTA inhibited the enzyme activity. The enzymatic hydrolyzed product of agar was characterized as neoagarobiose. Furthermore, this work is the first evidence of cold-adapted agarase in Antarctic psychrophilic bacteria and these results indicate the potential for the Antarctic agarase as a catalyst in medicine, food and cosmetic industries.

Purification and characterization of a thermostable λ-carrageenase from a hot spring bacterium, Bacillus sp.

Purpose of work The purpose of this study is to report a thermostable λ-carrageenase that can degrade λ-carrageenan yielding neo-λ-carrabiose at 75 °C.A thermophilic strain Lc50-1 producing λ-carrageenase was isolated from a hot spring in Indonesia and identified as a Bacillus sp. The λ-carrageenase, Cga-L50, with an apparent molecular weight of 37 kDa and a specific activity of 105 U/mg was purified from the culture supernatant. The optimum pH and temperature of Cga-L50 were 8.0 and 75 °C, respectively. The enzyme was stable from pH 6-9 and retained ~50 % activity after holding at 85 °C for 10 min. Significant activation of Cga-L50 was observed with K(+), Ca(2+), Co(2+), and Na(+); whereas, the enzyme activity was inhibited by Sr(2+), Mn(2+), Fe(2+), Cu(2+),Cd(2+), Mg(2+), and EDTA. Cga-L50 is an endo-type λ-carrageenase that hydrolyzes ß-1,4-linkages of λ-carrageenan, yielding neo-λ-carrabiose as the main product. This study is the first to present evidence of thermostable λ-carrageenase from hot spring bacteria.

Purification and characterization of thermostable agarase from Bacillus sp. BI-3, a thermophilic bacterium isolated from hot spring.

An extracellular agarase was purified from Bacillus sp. BI-3, a thermophilic agar-degrading bacterium isolated from a hot spring in Indonesia. The purified agarase revealed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular mass of 58 kDa. The optimum pH and temperature of the agarase were 6.4 and 70°C, respectively. The activity of the agarase was stable at high temperatures, and more than 50% activity was retained at 80°C for 15 min. Furthermore, the enzyme was stable in the pH range of 5.8-8.0, and more than 60% of the residual activity was retained. Significant activation of the agarase was observed in the presence of K(+), Na(+), Ca(2+), Mg(2+), and Sr(2+); on the other hand, Ba(2+), Zn(2+), Cu(2+), Mn(2+), Co(2+), Fe(2+), and EDTA inhibited or inactivated the enzyme activity. The components of the hydrolytic product analyzed by thin-layer chromatography showed that the agarase mainly produced neoagarobiose. This study is the first to present evidence of agarolytic activity in aerobic thermophilic bacteria.

[Analysis of the chemical constituents of essential oil from Ligustrum quihoui by GC-MS].

OBJECTIVE: [corrected] To study the chemical constituents of essential oil from Ligustrum quihoui. METHODS: Essential oil was extracted by steam distillation (SD). The chemical constituents of essential oil was analyzed by GC-MS. RESULTS: The chemical components in the oil were qualitatively and quantitatively analyzed by GC-MS, 76 components were seperated and 35 components were identified. The main components are n-Hexadecanoic acid (17.28%), (Z, Z, Z)-9, 12, 15-Octadecatrienoic acid, ethyl ester (12.13%), Phytol (5.80%). CONCLUSION: The method is simple, reliable and with good reprodutivity.