ONOO- Activatable Fluorescent Sulfur Dioxide Donor for a More Accurate Assessment of Cell Ferroptosis.

Ferroptosis is critical in the treatment of tumor therapies. Thus, monitoring reactive oxygen species (ROS) is of great significance for accurate assessment in ferroptosis without any interference. However, current probes for monitoring ROS during ferroptosis suffer from a drawback in that the probes consume ROS during detection, which inhibits the ferroptosis process and thus affects the accuracy and effectiveness of monitoring the process of ferroptosis. Herein, a new fluorescent donor probe, TFMU-SO2D, with the combination of the moiety of the SO2 donor is designed and synthesized by introducing the aryl boronate moieties that could give it the ability to effectively recognize ONOO-. The released SO2 could consume excess glutathione and regulate oxidative stress by elevating ROS levels, which would offset the ROS depletion by TFMU-SO2D and ensure accuracy in monitoring the ferroptosis process. The experimental results demonstrated that TFMU-SO2D possessed satisfactory performance for monitoring ONOO- as well as simultaneously releasing SO2 in oxidative stress stimulated by monensin and ferroptosis stimulated by erastin and RSL3. Additionally, the capability of SO2 synergized with ferroptosis to inhibit the viability of cancer cells was demonstrated by the CCK8 assay, which may be due to the fact that SO2 can potentiate ferroptosis cell death by increasing the ROS level. Overall, these combined results indicated that TFMU-SO2D possesses the excellent ability to precisely monitor ONOO- during ferroptosis without interference, which is significant for accurately accessing ferroptosis, cancer treatment, and drug development.

Evaluation of a novel lysis-based sample processing method to optimize Vibrio vulnificus detecting by loop-mediated isothermal amplification assay.

BACKGROUND: Vibrio vulnificus exists as one of the most serious foodborne pathogens for humans, and rapid and sensitive detection methods are needed to control its infections. As an emerging method, The Loop-Mediated Isothermal Amplification (LAMP) assay has been applied to the early detection of various foodborne pathogens due to its high efficiency, but sample preprocessing still prolongs the complete detection. To optimize the detection process, our study established a novel sample preprocessing method that was more efficient compared to common methods. RESULT: Using V. vulnificus as the detecting pathogen, the water-lysis-based detecting LAMP method shortened the preprocessing time to ≤ 1 min with 100% LAMP specificity; the detection limits of the LAMP assay were decreased to 1.20 × 102 CFU/mL and 1.47 × 103 CFU/g in pure culture and in oyster, respectively. Furthermore, the 100% LAMP specificity and high sensitivity of the water-lysis method were also obtained on detecting V. parahaemolyticus, V. alginolyticus, and P. mirabilis, revealing its excellent LAMP adaption with improvement in sensitivity and efficiency. CONCLUSION: Our study provided a novel LAMP preprocessing method that was more efficient compared to common methods and possessed the practical potential for LAMP application in the future.

Birefringence characterization in a dual-hole microstructured optical fiber using an OFDR method.

The birefringence in a dual-hole microstructured optical fiber is numerically calculated and characterized with an optical frequency domain reflectometry (OFDR) method. Due to the asymmetric dual air holes in the cross-section, the polarized L P01x and L P01y modes propagate with different group velocities and time delays. Through a polarized coherent OFDR system in experiment, the Fresnel reflection peaks for each mode are separated in the frequency domain with their corresponding beat frequency. Thus, the group birefringence -9.68×10-4 is calculated with a beat frequency difference of 50.03 Hz between the L P01x and L P01y modes at a 6.2 m fiber end, which is in good agreement with that of -9.54×10-4 from the theoretical simulation. Our demonstration provides an accurate and flexible method for group birefringence characterization in microstructured optical fibers with complex cross-sectional structures.

Association between lactate/albumin ratio and prognosis in critically ill patients with acute kidney injury undergoing continuous renal replacement therapy.

BACKGROUND: The primary objective was to examine the association between the lactate/albumin ratio (LAR) and the prognosis of patients with acute kidney injury (AKI) undergoing continuous renal replacement therapy (CRRT). METHODS: Utilizing the Medical Information Mart for Intensive Care IV (MIMIC-IV, v2.0) database, we categorized 703 adult AKI patients undergoing CRRT into survival and non-survival groups based on 28-day mortality. Patients were further grouped by LAR tertiles: low (< 0.692), moderate (0.692-1.641), and high (> 1.641). Restricted cubic splines (RCS), Least Absolute Shrinkage and Selection Operator (LASSO) regression, inverse probability treatment weighting (IPTW), and Kaplan-Meier curves were employed. RESULTS: In our study, the patients had a mortality rate of 50.07% within 28 days and 62.87% within 360 days. RCS analysis revealed a non-linear correlation between LAR and the risk of mortality at both 28 and 360 days. Cox regression analysis, which was adjusted for nine variables identified by LASSO, confirmed that a high LAR (>1.641) served as an independent predictor of mortality at these specific time points (p < 0.05) in AKI patients who were receiving CRRT. These findings remained consistent even after IPTW adjustment, thereby ensuring a reliable and robust outcome. Kaplan-Meier survival curves exhibited a gradual decline in cumulative survival rates at both 28 and 360 days as the LAR values increased (log-rank test, χ2 = 48.630, p < 0.001; χ2 = 33.530, p < 0.001). CONCLUSION: A high LAR (>1.641) was found to be an autonomous predictor of mortality at both 28 and 360 days in critically ill patients with AKI undergoing CRRT.

Imine Bond-Based Fluorescent Nanofilms toward High-Performance Detection and Efficient Removal of HCl and NH3.

A new kind of imine bond-based fluorescent nanofilm was developed as multifunctional materials for high-performance detection and efficient removal of hydrogen chloride (HCl) and ammonia (NH3). The flexible, uniform, and photochemically stable nanofilms as prepared showed fast (<1 and <0.5 s), sensitive (<150 ppb and <1.5 ppm), and selective response to HCl and NH3, respectively, and the removal efficiencies to HCl and NH3 are 187.5 and 37.5% (w/w), respectively. A reversible earthy-red to green fluorescence color change upon adsorption of NH3 or HCl enabled visualized monitoring of the two gases in the air. Mechanism studies revealed that the adsorption of HCl is a result of hydrogen bond formation between the analyte and the imine groups. Adsorption of NH3, however, is a result of chemical reaction with the pre-adsorbed HCl. The applicability of the detection and removal strategies as developed was further verified by conducting the tests on real-life or simulated scenarios.

The effects of polyethylene microplastics on the growth, reproduction, metabolic enzymes, and metabolomics of earthworms Eisenia fetida.

The existing data regarding the effects of polyethylene (PE) microplastics (MPs) smaller than 5 mm in size on earthworms are insufficient to fully comprehend their toxicity. In this study, earthworms Eisenia fetida were exposed to artificially added PE at a concentration ranging from 0.05 to 20 g/kg soil (0.005%-2%) for 60 days to determine the concentration range causing negative effects on earthworms and to uncover the potential toxic mechanisms. The individual growth, reproduction, and metabolic enzyme activities, including phase I enzymes (cytochrome P450 [CYP] 1A2, 2B6, 2C9, and 3A4), and phase II metabolic enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione sulfotransferase (GST)), and metabolomics were measured. The observed variations in responses of multiple cross-scale endpoints indicated that individual indices are less responsive to PE MPs than metabolic enzymes or metabolomics. Despite the absence of significant alterations in growth inhibition based on body weight, PE MPs at concentrations equal to or exceeding 2.5 g/kg were found to exert a toxic effect on earthworms, which was evidenced by significant changes in metabolic enzyme activities (CYP1A2, 2B6, 2C9, and 3A4, SOD, CAT, and GST) and important small molecule metabolites screened based on metabolomics, likely due to the bioaccumulation of PE. The toxicity of PE MPs to earthworms is inferred to be associated with neurotoxicity, oxidative damage, decreased detoxification capacity, energy metabolism imbalance, and impaired amino acid and purine metabolism due to bioaccumulation. The findings of this study will enhance our understanding of the molecular toxicity mechanisms of PE MPs and contribute to a more accurate assessment of the ecological risks posed by PE MPs in soil.

Observation and characterization of the high order modes in a six-mode fiber using an OFDR method.

High order modes in a six-mode fiber are separately observed and characterized using an Optical Frequency Domain Reflectometry (OFDR) method. Due to the difference in group refractive index between fundamental mode and the high order modes, Fresnel reflection peaks for each mode can be separated in beat frequency domain with their corresponding time delay. In the experiment, the fundamental mode and high order modes are excited in turn and observed at a 6.6 m six-mode fiber end, which agree with their beat frequency difference in theoretical simulation. The demonstration provides a flexible and feasible method for mode identification and characterization of all kinds of fibers.

A model for continuous sample feed and separation with counter-current chromatography based on elution-extrusion mode and its application.

A novel and meaningful theoretical model was established with counter-current chromatography based on the elution-extrusion mode for efficient continuous separation. For the experimental verification of the theory, the separation of the binary mixture luteolin/baicalein was studied. The velocity model and volume model of the chromatographic separation behavior of the target compounds in the separation process were given by theoretical analysis. The results showed that this method had obvious advantages in the separation of binary mixtures. In addition, the established model was used to predict and isolate oleuropein from olive leaves. A two-phase solvent system of n-butanol/ethyl acetate/methanol/water (1:19:1:19, v/v/v/v) was chosen for the continuous separation of oleuropein. After optimizing the conditions in this way, a large amount of sample loading was achieved; the volume of injections can reach 48 ml, approximately 35.29% of the volume of the counter-current chromatography column, and oleuropein with a purity of 86.42% was obtained within 80 min. The model provided technical support for the prediction of chromatographic behavior and operating parameters during continuous separation and preparation of counter-current chromatography. It has great application prospects and significance in separation preparation, especially in large-scale industrial preparation.

Effects of multi-walled carbon nanotubes in soil on earthworm growth and reproduction, enzymatic activities, and metabolomics.

Increased production and environmental release of multi-walled carbon nanotubes (MWCNTs) increase soil exposure and potential risk to earthworms. However, MWCNT toxicity to earthworms remains unclear, with some studies identifying negative effects and others negligible effects. In this study, to determine whether exposure to MWCNTs negatively affects earthworms and to elucidate possible mechanisms of toxicity, earthworms were exposed to sublethal soil concentrations of MWCNTs (10, 50, and 100 mg/kg) for 28 days. Earthworm growth and reproduction, activities of cytochrome P450 (CYP) isoforms (1A2, 2C9, and 3A4) and antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione-s-transferase (GST)), and metabolomics were determined. Effects of MWCNTs on earthworms depended on exposure concentration. Exposure to MWCNTs did not significantly affect growth and reproduction of individual earthworms. Exposure to 50 mg/kg MWCNTs significantly increased activities of CYP2C9, CYP3A4, SOD, CAT, and GST but clearly reduced levels of L-aspartate, L-asparagine, and glutamine. With exposure to 100 mg/kg MWCNTs, toxic effects on earthworms were observed, with significant inhibition in activities of CYP isoenzymes and SOD, significant reductions in L-aspartate, L-asparagine, glutamine, and tryptophan, and simultaneous accumulations of citrate, isocitrate, fumarate, 2-oxoglutarate, pyruvate, D-galactose, carbamoyl phosphate, formyl anthranilate, hypoxanthine, and xanthine. Results suggest that toxicity of MWCNTs to earthworms is associated with reduced detoxification capacity, excessive oxidative stress, and disturbance of multiple metabolic pathways, including amino acids metabolism, the tricarboxylic acid cycle, pyruvate metabolism, D-galactose metabolism, and purine metabolism. The study provides new insights to better understand and predict the toxicity of MWCNTs in soil.

A long-wavelength activable AIEgen fluorescent probe for HClO and cell apoptosis imaging.

Hypochlorous acid (HClO) is an important bactericide, and adjusting the content of HClO helps to improve the host's innate immunity and resist microbial invasion. Aggregation-induced luminescence (AIE) is the opposite of aggregation-induced quenching (ACQ). Compounds with AIE properties emit weakly in a dispersed state in solution and they can emit strong fluorescence in an aggregated state. In this article, we proposed a new AIE fluorescent probe QM-ClO based on the quinoline-malononitrile (QM) fluorophore and dimethylthiocarbamate (DMTC) to detect HClO. The probe QM-ClO showed a fast response time, a low detection limit of 30.8 nM and a large Stokes shift (190 nm). Carbonyl cyanide metachlorophenyl-hydrazone (CCCP) was used to induce cell apoptosis, and then an increase in the HClO content was observed in the cell. It is proved that cell apoptosis can lead to the increase of the HClO content in the cell. This probe provides an effective tool for studying apoptosis-related diseases.

A robust, freeze-resistant and highly ion conductive ionogel electrolyte towards lithium metal batteries workable at -30 °C.

The wide applications of lithium metal batteries have encountered a severe conductivity issue when operating in cold weather. Here we report a freeze-resistant lithium metal battery, which displays outstanding rate performance, negligible polarization deterioration, and a good capacity retention of 94.25% after 700-cycles of use at -30 °C, the lowest temperature ever reported for gel electrolyte-based lithium metal batteries. Remarkably, the lithium metal batteries are even workable at temperatures down to -60 °C. The key point of the innovative design is the utilization of a newly created anti-freezing ionogel as an electrolyte, which is produced by gelation of an electrochemically inert ionic liquid, 1-butyl-3-methylimidazolium tetrafluoro-borate ([BMIM]BF4), via dynamic condensation of a specially designed benzaldehyde-terminated polyethylene glycol (PEG-CHOs) with the tetra-hydrazide derivative of p-tert-butyl-calix[4]arene (CTH). The as-prepared ionogel electrolyte demonstrates a high ionic conductivity (0.43 mS cm-1), a broad stability window (2.4-4.3 V vs. Li+/Li), and high flexibility at -30 °C. The outstanding property of the ionogel electrolyte is ascribed to its unique gel network structure as it enables enrichment of Li+ and enhances its efficient transportation. Further tests demonstrate that the ionogel electrolyte could be also used for the assembly of flexible lithium metal batteries.

Recovery and recycling of solvent of counter-current chromatography: The sample of isolation of zeaxanthin in the Lycium barbarum L. fruits.

An efficient method of recovering and recycling solvent for counter-current chromatography was established by which zeaxanthin was separated from Lycium barbarum L. fruits. A column with activated carbon combined with high performance counter-current chromatography formed the recovering and recycling solvent system. Using the solvent system of n-hexane-ethyl acetate-ethanol-water (8:2:7:3, v/v) from the references, five injections were performed with an almost unchanged purity of zeaxanthin (80.9, 81.2, 81.5, 81.3, and 80.2% respectively) in counter-current chromatography separation. Meanwhile, the mobile phase reduced by half than conventional counter-current chromatography. By this present method, an effective improvement of counter-current chromatography solvent utilization was achieved.

Evaluation of the combined toxicity of multi-walled carbon nanotubes and cadmium on earthworms in soil using multi-level biomarkers.

The coexistence of multi-walled carbon nanotubes (MWCNTs) with cadmium (Cd) in soil may cause the combined biological effects, but few study reported about their joint toxic effects on earthworms. Therefore, this study investigated the effects of sub-lethal levels of MWCNTs (10, 50, 100 mg/kg) and Cd (2.0, 10 mg/kg) on earthworms Eisenia fetida for 14 days. The changes in multi-level biomarkers of growth inhibition rate, cytochrome P450 isoenzymes (CYP1A2, 2C9 and 3A4), and small molecular metabolites (metabolomics) were determined. The toxic interaction between MWCNTs and Cd was characterized by the combination of the biomarker integration index (BRI), joint effect index concentration addition index (CAI), and the effect concentration addition index (EAI). The results showed that the single MWCNTs exposure caused insignificant change in most biomarkers, while the combined exposure of MWCNTs (50-100 mg/kg) and 10 mg/kg Cd led to significant changes in ten most important metabolites identified by metabolomics and activities of CYP1A2, 2C9, and 3A4. Compared with the toxicity of Cd alone, the combined toxicity of the mixture was significantly reduced. According to the integration of BRI and CAI/EAI, a clearly antagonistic interaction at relatively low effects was observed between MWCNTs and Cd. The responses of multiple biomarkers suggest the toxic action mode of the mixture on earthworms was related to the oxidative injury, and the disruption of amino acid, purine, and pyrimidine metabolism, and the urea cycle.

The responses of the growth, cytochrome P450 isoenzymes activities and the metabolomics in earthworms to sublethal doses of dichlorvos in soil.

In this paper, earthworms (Eisenia fetida) were exposed to sublethal doses of dichlorvos (spiked concentration of 0.1, 1.0, 10 mg/kg) in soil for 14 days, the metabolomics and activities of cytochrome P450 (CYP) isoenzymes (CYP1A2, CYP2C9 and CYP3A4) of earthworms were analyzed aiming to identify sensitive biomarkers and reveal possible mode of toxic action. The results showed that CYP1A2 and CYP2C9 activity appeared to be more sensitive than CYP3A4 activity in response to dichlorvos, and that metabolic responses based on the metabolomics depended on both of the length of exposure and exposure dose. Malate, ornithine, glucose, inosine, myo-inositol and some amino acids (glutamine, tryptophan, phenylalanine, tyrosine, leucine, histidine, glutamate, lysine) and CYP isozenzymes may be biomarkers to reveal the toxic effect of dichlorvos on earthworms. Compared to controls, when dichlorvos dose reached 1.0 and 10 mg/kg on day 14, glucose and ornithine increased significantly, malate and some amino acids (glutamine, tryptophan, phenylalanine, tyrosine, leucine) decreased significantly, and activities of CYP1A2 and CYP2C9 were inhibited significantly. The current results suggested that 1.0 and 10 mg/kg dichlorvos for 14 days of exposure blocked energy metabolism, disordered Krebs cycle, interfered amino acids metabolism and evoked toxic effects on earthworms.

Development of a Column-Shaped Fluorometric Sensor Array and Its Application in Visual Discrimination of Alcohols from Vapor Phase.

Portable, miniaturized, and inexpensive detectors are in high demand for detecting and discriminating volatile organic compounds (VOCs). Sensor array design and exploitation are two key issues for new detector development. In contrast to the most reported plane-shaped sensor array for gaseous analyte sensing, here we report a column-shaped fluorometric sensor array by using fluorophore-loaded silica particles (∼40 µm) filled capillary. In the design, the capillary serves as test chamber and facilitates visualization. The orifices of the capillary were used as inlet and outlet for gaseous analyte. Sensing modules are installed in series, which lays foundation for their even and effective contact with the gaseous analyte. Meanwhile, further capsulation could be avoided. Silica particles were chosen as carries due to their preferred adsorption behavior to VOCs. By choosing four typical fluorophores (PBI-CB, Py-CB-Ph, Py-At, and NA-Ch) as sensing units, a 4-element fluorometric sensor array was achieved. Fluorescence of the array varied when different alcohol vapors were pumped in. The six tested alcohols could not only be distinguished as primary, secondary, or tertiary, but also be identified individually. The array had good reproducibility in visualization of the six alcohols. In addition, the orders of the fluorophores can be changed as desired. It is believed that the proofed concept provides not only a totally new design of sensor array but also contributes a new strategy for the discrimination of the alcohols as examined.

Imaging Hg2+-Induced Oxidative Stress by NIR Molecular Probe with "Dual-Key-and-Lock" Strategy.

Mercury (Hg) is considered an extremely toxic heavy metal which is extremely harmful to both the human body and environment. In addition, Hg2+-induced oxidative stress also exerts a crucial role to play in pathophysiological mechanisms of mercury toxicity. Thus, efficient and specific fluorescent probes for imaging Hg2+-induced oxidative stress are necessary. In the present study, we rationally design a novel Hg2+-activated and ICT-based NIR emission fluorescent probe NIR-HO for sequentially monitoring the ONOO- level with a "dual-key-and-lock" strategy. The probe NIR-HO showed rapid response and excellent specificity and sensitivity for the detection of Hg2+ and ONOO- in vitro. Cell imaging demonstrated that Hg2+-induced oxidative stress was involved in ONOO- upregulation. Also, GSH, NAC, and EDTA were employed as excellent detoxifying drugs against Hg2+-induced toxicity. Moreover, the probe NIR-HO was successfully used for imaging Hg2+ and ONOO- in vivo. In brief, NIR-HO provides a simple and powerful approach which can be used to image Hg2+-induced oxidative stress in the pathological environment.

Separation and purification of hydroxytysol and oleuropein from Olea europaea L. (olive) leaves using macroporous resins and a novel solvent system.

The separation and purification of hydroxytysol and oleuropein from Olea europaea L. (olive) using a macroporous resin with a novel solvent system was systematically investigated. Static adsorption experiments with BMKX-4 resin revealed that the experimental data of both hydroxytysol and oleuropein fitted best to the pseudo-second-order kinetic and Freundlich isotherm models. The thermodynamic parameters indicated spontaneous and exothermic adsorption processes. The novel solvent system, composed of n-hexane:ethyl acetate:methanol:water in a (v/v/v/v) ratio of 1:9:1:9, had two phases (upper and lower). The separation and purification parameters of hydroxytysol and oleuropein were optimized using dynamic adsorption/desorption on a column packed with BMKX-4 resin. The effects of flow rates and volumes of the upper and lower phases on the separation efficiency were systematically studied. Under optimal conditions, the fraction of hydroxytysol in the final product increased by 6.34-fold from 0.46 to 2.96%, with a yield rate of 88.58% w/w, while that of oleuropein increased 4.17-fold from 11.40 to 47.59%, with a 93.31% w/w yield rate. These results may be help in selecting a suitable eluent for improved separation of macroporous adsorption resins.

Full use of the liquid nature of the stationary phase: The development of elution-extrusion counter current chromatography.

Elution-extrusion counter current chromatography extrudes the most solute retained in the column with the highest possible peak resolution. It can greatly improve the hydrophobic window. In recent years, elution-extrusion counter current chromatography has received extensive attention in the separation of complex samples. This article first reviews the development and application of elution-extrusion counter current chromatography, including its origin, mechanism, advantages and disadvantages, and some representative applications. At the same time, this review also shared our visions and ideas on how to improve the elution-extrusion mode. This article aims to provide certain reference for the research of this technology.

Effective on-line high-speed shear dispersing emulsifier technique coupled with high-performance countercurrent chromatography method for simultaneous extraction and isolation of carotenoids from Lycium barbarum L. fruits.

An efficient combination strategy based on high-speed shear dispersing emulsifier technique and high-performance countercurrent chromatography was developed for on-line extraction and isolation of carotenoids from the fruits of Lycium barbarum. In this work, the high-speed shear dispersing emulsifier technique has been employed to extract crude extracts using the upper phase of high-performance countercurrent chromatography solvent system composed of n-hexane-dichloromethane-acetonitrile (10:4:6.5, v/v) as the extraction solvent. At the separation stage, the high-performance counter-current chromatography process adopts elution-extrusion mode and the upper phase of the solvent system as stationary phase (reverse-phase mode). As a result, three compounds including zeaxanthin, zeaxanthin monopalmitate, and zeaxanthin dipalmitate with purities of 89, 90, and 93% were successfully obtained in one extraction-separation operation within 120 min. The targeted compounds were analyzed and identified by high-performance liquid chromatography, mass spectrometry, and NMR spectroscopy. The results indicated that the present on-line combination method could serve as a simple, rapid, and effective way to achieve weak polar and unstable compounds from natural products.

Numerical Analysis and Recursive Compensation of Position Deviation for a Sub-Millimeter Resolution OFDR.

We analyze the source of the position deviation and propose a demodulation recursive compensation algorithm to ensure a sub-millimeter resolution in improved optical frequency domain reflectometry. The position deviation between the geometric path and optical path changes with the temperature or strain, due to the elastic-optic and thermal-optic effects. It accumulates along the fiber and becomes large enough to affect the spectral correlation between the measured and reference spectra at the fiber end. The proposed algorithm compensates for the position deviation of each measuring point and aligns the measured spectra with its reference. The numerical and experimental results both reveal that the signal-to-noise ratio of the correlation is improved doubly and a sub-millimeter spatial resolution becomes available at a 30 m fiber end. The recursive compensation algorithm helps to restrain the correlation degeneration at the fiber end and promises an effective approach to a sub-millimeter resolution in optical frequency domain reflectometry.