Herein we have reported a fluorescent probe (MB-M) based on MB derivative for Cu2+ ions detection. The probe was well characterized by 1H NMR, 13C NMR and HR-MS spectrum. Probe MB-M showed naked-eyes recognition to Cu2+ as color change from colorless to indigo. The probe exhibited promising features such as high fluorescence and UV-vis selectivity, fast response (5 mint), workable at pH 2-7, and low limit of detection (LOD = 0.33 µM). Probe MB-M was also used for Cu2+ ions imaging in HepG-2 cells and detection in daily life (Test Strip and lake water). Moreover, non-covalent interaction (NCI) and quantum theory of atoms in molecules (QTAIM) analysis were used to study the interaction between MB-M and Cu2+ ions. By examining the electronic characteristics of the complex using natural bond orbital (NBO), electron density difference (EDD), and frontier molecular orbital (FMO) analysis, the sensitivity of MB-M towards Cu2+ ions were investigated. The results illustrated that the interactions between MB-M and Cu2+ ions involved chemisorption.

This review provides a thorough examination of small molecule-based fluorescence chemosensors tailored for bioimaging applications, showcasing their unique ability to visualize biological processes with exceptional sensitivity and selectivity. It explores recent advancements, methodologies, and applications in this domain, focusing on various designs rooted in anthracene, benzothiazole, naphthalene, quinoline, and Schiff base. Structural modifications and molecular engineering strategies are emphasized for enhancing sensor performance, including heightened sensitivity, selectivity, and biocompatibility. Additionally, the review offers valuable insights into the ongoing development and utilization of these chemosensors, addressing current challenges and charting future directions in this rapidly evolving field.

Synchronous fluorescence spectroscopy (SFS) is a technique that involves the simultaneous detection of fluorescence excitation and emission at a constant wavelength difference. The spectrum yields bands that are narrower and less complex than the original excitation and emission bands. The SFS bands correspond uniquely to the fluorescing molecule. Our investigation focuses on evaluating the sensitivity of the SFS technique for the detection and quantitation of PAHs relevant to astrochemistry. Results are presented for naphthalene, anthracene, and pyrene in three different solvents: n-hexane, water, and ethanol. SF bands are obtained with a constant wavelength difference between the peak excitation and emission wavelength (Δλ = λex - λem) at a concentration ranging from 10-4 to 10-10 M. Limit of detection (LOD) and limit of quantitation (LOQ) calculations are based on integrated SF band areas at different concentrations. Spectra of 23 pg/g of anthracene, 16 pg/g, and 2.6 pg/g of pyrene are recorded using ethanol as the solvent. The PAHs exhibit detection limits in the fractions of parts-per-billion (ng/g) range. Through comparison with similar prior studies employing fluorescence emission, our findings reveal a better detectability limit, demonstrating the effectiveness and applicability of the SFS technique.

The recent regulatory spotlight on continuous monitoring (CM) solutions and the rapid development of CM solutions have demanded the characterization of solution performance through regular, rigorous testing using consensus test protocols. This study is the second known implementation of such a protocol involving single-blind controlled testing of 9 CM solutions. Controlled releases of rates (6-7100 g) CH4/h over durations (0.4-10.2 h) under a wind speed range of (0.7-9.9 m/s) were conducted for 11 weeks. Results showed that 4 solutions achieved method detection limits (DL90s) within the tested emission rate range, with all 4 solutions having both the lowest DL90s (3.9 [3.0, 5.5] kg CH4/h to 6.2 [3.7, 16.7] kg CH4/h) and false positive rates (6.9-13.2%), indicating efforts at balancing low sensitivity with a low false positive rate. These results are likely best-case scenario estimates since the test center represents a near-ideal upstream field natural gas operation condition. Quantification results showed wide individual estimate uncertainties, with emissions underestimation and overestimation by factors up to >14 and 42, respectively. Three solutions had >80% of their estimates within a quantification factor of 3 for controlled releases in the ranges of [0.1-1] kg CH4/h and > 1 kg CH4/h. Relative to the study by Bell et al., current solutions performance, as a group, generally improved, primarily due to solutions from the study by Bell et al. that were retested. This result highlights the importance of regular quality testing to the advancement of CM solutions for effective emissions mitigation.

Glucaric acid (GA) is a value-added chemical and can be used to manufacture food additives, anticancer drugs, and polymers. The non-genetic cell-to-cell variations in GA biosynthesis are naturally inherent, indicating the presence of both high- and low-performance cells in culture. Low-performance cells can lead to nutrient waste and inefficient production. Furthermore, myo-inositol oxygenase (MIOX) is a key rate-limiting enzyme with the problem of low stability and activity in GA production. Therefore, eliminating cell-to-cell variations and increasing MIOX stability can select high-performance cells and improve GA production. In this study, an in vivo GA bioselector was constructed based on GA biosensor and tetracycline efflux pump protein TetA to continuously select GA-efficient production strains. Additionally, the upper limit of the GA biosensor was improved to 40 g/L based on ribosome-binding site optimization, achieving efficient enrichment of GA high-performance cells. A small ubiquitin-like modifier (SUMO) enhanced MIOX stability and activity. Overall, we used the GA bioselector and SUMO-MIOX fusion in fed-batch GA production and achieved a 5.52-g/L titer in Escherichia coli, which was 17-fold higher than that of the original strain.IMPORTANCEGlucaric acid is a non-toxic valuable product that was mainly synthesized by chemical methods. Due to the problems of non-selectivity, inefficiency, and environmental pollution, GA biosynthesis has attracted significant attention. The non-genetic cell-to-cell variations and MIOX stability were both critical factors for GA production. In addition, the high detection limit of the GA biosensor was a key condition for performing high-throughput screening of GA-efficient production strains. To increase GA titer, this work eliminated the cell-to-cell variations by GA bioselector constructed based on GA biosensor and TetA, and improved the stability and activity of MIOX in the GA biosynthetic pathway through fusing the SUMO to MIOX. Finally, these approaches improved the GA production by 17-fold to 5.52 g/L at 65 h. This study represents a significant step toward the industrial application of GA biosynthetic pathways in E. coli.

The Flinders Technology Associates (FTA) card, a cotton-based cellulose membrane impregnated with a chaotropic agent, effectively inactivates infectious microorganisms, lyses cellular material, and fixes nucleic acid. The aim of this study is to assess the stability and detection limit of various RNA viruses, especially the avian influenza virus (AIV), Newcastle disease virus (NDV), and African horse sickness virus (AHSV), on the FTA card, which could significantly impact virus storage and transport practices. To achieve this, each virus dilution was inoculated onto an FTA card and stored at room temperature in plastic bags for durations ranging from 1 week to 6 months. Following storage, the target genome was detected using conventional reverse transcription polymerase chain reaction. The present study demonstrated that the detection limit of AIV ranged from 1.17 to 6.17 EID50 values over durations ranging from 1 week to 5 months, while for NDV, it ranged from 2.83 to 5.83 ELD50 over the same duration. Additionally, the detection limit of AHSV was determined as 4.01 PFU for both 1 and 2 weeks, respectively. Based on the demonstrated effectiveness, stability, and safety implications observed in the study, FTA cards are recommended for virus storage and transport, thus facilitating the molecular detection and identification of RNA viral pathogens.

Due to the toxicity of mercury and its harmful effects on human health, it is essential to establish a low-cost, highly sensitive and highly specific monitoring method with a wide detection range, ideally with a simple visual readout. In this study, a whole-cell biosensor with adjustable detection limits was developed for the detection of mercury ions in water samples, allowing controllable threshold detection with an expanded detection range. Gene circuits were constructed by combining the toehold switch system with lactose operon, mercury-ion-specific operon, and inducible red fluorescent protein gene. Using MATLAB for design and selection, a total of eleven dual-input single-output sensing logic circuits were obtained based on the basic logic of gene circuit construction. Then, biosensor DTS-3 was selected based on its fluorescence response at different isopropyl β-D-Thiogalactoside (IPTG) concentrations, exhibiting the controllable detection threshold. At 5-20 μM IPTG, DTS-3 can achieve variable threshold detection in the range of 0.005-0.0075, 0.06-0.08, 1-2, and 4-6 μM mercury ion concentrations, respectively. Specificity experiments demonstrated that DTS-3 exhibits good specificity, not showing fluorescence response changes compared with other metal ions. Furthermore spiked sample experiments demonstrated its good resistance to interference, allowing it to distinguish mercury ion concentrations as low as 7.5 nM by the naked eye and 5 nM using a microplate reader. This study confirms the feasibility and performance of biosensor with controllable detection threshold, providing a new detection method and new ideas for expanding the detection range of biosensors while ensuring rapid and convenient measurements without compromising sensitivity.

UNASSIGNED: Difenoconazole (DIFE) is a common pesticide used in citrus cultivation; excessive intake can cause neurological damage to the organism, and the existing colloidal gold immunochromatographic test strips cannot meet the requirements for the detection of citrus samples.
UNASSIGNED: Difenoconazole test strip was prepared based on the colloidal gold immunochromatographic technique (GICT), and its application in citrus samples was investigated; with colloidal gold (CG) as the probe, the optimization of GICT parameters, and the determination of reaction method, the immunochromatographic test strips for the detection of DIFE in citrus was developed, and the limit of detection (LOD), specificity, accuracy, and stability of the test strips were verified.
UNASSIGNED: The results showed that the visual detection limit of the prepared colloidal gold immunochromatographic test strips was 0.2 mg/kg and the quantitative range was 0.06-0.6 mg/kg, and the test strips could specifically identify DIFE and have no cross-reaction with other common triazole pesticides. The detection method established in this study was verified by the GC-MS method, and the detection results achieved good consistency (R2 > 0.98).
UNASSIGNED: The test strips developed in this study have good performance and can be used for highly sensitive detection of citrus samples.

CA125 (carbohydrate antigen 125) is an important biomarker of ovarian cancer, so developing effective method for its detection is of great significance. In the present work, a novel sandwich-like electrochemical immunosensor (STEM) of CA125 was constructed by preparing nanoribbon-like Ti3C2Tx MXenes (Ti3C2TxNR) to immobilize primary antibody (PAb) of CA125 and UIO-66-NH2 MOFs structure to immobilize second antibody (SAb) and electroactive toluidine blue (Tb) probe. In this designed STEM assay, the as-prepared Ti3C2TxNR nanohybrid offers the advantages in large surface area and conductivity as carrier, and UIO-66-NH2 provided an ideal platform to accommodate SAb and a large number of Tb molecules as signal amplifier. In the presence of CA125, the peak currents of Tb from the formed STEM structure increase with the increase of CA125 level. After optimizing the related control conditions, a wide linear range (0.2-150.0 U mL-1) and a very low detection limit (0.05 U mL-1) of CA125 were achieved. It\'s thus expected the developed STEM strategy has important applications for the detection of CA125.

UNASSIGNED: Acute haemorrhagic diarrhoea syndrome (AHDS) in dogs is a condition of unknown aetiology. Providencia alcalifaciens is suspected to play a role in the disease as it was commonly found in dogs suffering from AHDS during a Norwegian outbreak in 2019. The role of this bacterium as a constituent of the canine gut microbiota is unknown, hence this study set out to investigate its occurrence in healthy dogs using metagenomics.
UNASSIGNED: To decrease the likelihood of false detection, we established a metagenomic threshold for P. alcalifaciens by spiking culture-negative stool samples with a range of bacterial dilutions and analysing these by qPCR and shotgun metagenomics. The detection limit for P. alcalifaciens was determined and used to establish a metagenomic threshold. The threshold was validated on naturally contaminated faecal samples with known cultivation status for P. alcalifaciens. Finally, the metagenomic threshold was used to determine the occurrence of P. alcalifaciens in shotgun metagenomic datasets from canine faecal samples (n=362) collected in the HUNT One Health project.
UNASSIGNED: The metagenomic assay and qPCR had a detection limit of 1.1x103 CFU P. alcalifaciens per faecal sample, which corresponded to a Cq value of 31.4 and 569 unique k-mer counts by shotgun metagenomics. Applying this metagenomic threshold to 362 faecal metagenomic datasets from healthy dogs, P. alcalifaciens was found in only 1.1% (95% CI [0.0, 6.8]) of the samples, and then in low relative abundances (median: 0.04%; range: 0.00 to 0.81%). The sensitivity of the qPCR and shotgun metagenomics assay was low, as only 40% of culture-positive samples were also positive by qPCR and metagenomics.
UNASSIGNED: Using our detection limit, the occurrence of P. alcalifaciens in faecal samples from healthy dogs was low. Given the low sensitivity of the metagenomic assay, these results do not rule out a significantly higher occurrence of this bacterium at a lower abundance.