A Copper (II) Ensemble-Based Fluorescence Chemosensor and Its Application in the 'Naked-Eye' Detection of Biothiols in Human Urine.

Quick and effective detection of biothiols in biological fluids has gained increasing attention due to its vital biological functions. In this paper, a novel reversible fluorescence chemosensor (L-Cu2+) based on a benzocoumarin-Cu2+ ensemble has been developed for the detection of biothiols (Cys, Hcy and GSH) in human urine. The chemosensing ensemble (L-Cu2+) contains a 2:1 stoichiometry structure between fluorescent ligand L and paramagnetic Cu2+. L was found to exclusively bond with Cu2+ ions accompanied with a dramatic fluorescence quenching maximum at 443 nm and an increase of an absorbance band centered at 378 nm. Then, the in situ generated fluorescence sluggish ensemble, L-Cu2+, was successfully used as a chemosensor for the detection of biothiols with a fluorescence "OFF-ON" response modality. Upon the addition of biothiols, the decomplexation of L-Cu2+ led to the liberation of the fluorescent ligand, L, resulting in the recovery of fluorescence and absorbance spectra. Studies revealed that L-Cu2+ possesses simple synthesis, excellent stability, high sensitivity, reliability at a broad pH range and desired renewability (at least 5 times). The practical application of L-Cu2+ was then demonstrated by the detection of biothiols in human urine sample.

Two D-π-A Schiff-Base-Functionalized Silica Gel Adsorbents for Preconcentration of Copper Ions in Foods and Water for Detection.

Copper is an essential element in many biological processes and plays an important role in carbohydrate and lipid metabolism. Excess or deficiency of Cu ions can cause disturbances in cellular homeostasis and damage the central nervous system. Here, for the first time, two functionalized silica gel (SG-A and SG-B) adsorbents were prepared and tested for copper detection via the reactions of chlorinated silica gel with two novel D-π-A Schiff base compounds: 2-amino-3-(quinolin-2-ylmethyleneamino)maleonitrile (A) and 2-(4-(diethylamino)-2-hydroxybenzylideneamino)-3-aminomaleonitrile (B) in the thionyl chloride solution, respectively. SG-A and SG-B as adsorbents filled in a microcolumn were used to enrich trace Cu ions in foods and water with the detection of flame atomic absorption spectrometry. Because of the strong coordination between two D-π-A Schiff base compounds and Cu2+ ions, the stable heterocyclic Cu2+-SG-A/B complex is formed. For a sample volume of 30 mL, detection limits of 0.09 µg L-1 and 0.15 µg L-1 have been achieved. The results of selectivity study show that the two adsorbents can selectively extract Cu2+ in complex matrixes with other metal cations. The methods have been successfully applied to the determination of Cu2+ content in various real samples, and the detection sensitivity that we report here is better than most results reported using modified silica gels.

A fast response fluorescence probe specific for hypochlorous acid detection and its applications in bioimaging.

Quantitative detection and visualization of hypochlorous acid (HOCl) in biological samples has emerged as a hot topic in biochemical research because of the important role of this biomolecule in live organisms. In this contribution, a novel 1,8-naphthalimide-based fluorescence probe, DNPH-NA, was designed and synthesized for HOCl detection in aqueous solutions and in biological systems. In the presence of HOCl, significant changes in absorption and fluorescence spectra were observed, which allow for the quantitative detection of HOCl in PBS buffer. The detection limit for HOCl was determined to be 50 nM. In the presence of HOCl, the fluorescence response of DNPH-NA was found to be completed within 2 seconds. The desirable features of DNPH-NA for the detection of HOCl in aqueous solutions, such as its high sensitivity and selectivity, reliability at physiological pH, rapid fluorescence response, and biocompatibility enabled its application in the detection of HOCl in biological samples. Imaging of exogenous HOCl in live HeLa cells and endogenous HOCl generation in live J774A.1 macrophage cells and zebrafish was then successfully performed using DNPH-NA as a fluorescence probe.

Quantification of Gly m 5.0101 in Soybean and Soy Products by Liquid Chromatography-Tandem Mass Spectrometry.

Gly m 5.0101, the alpha subunit of ß-conglycinin, is one of the major allergens found in soybeans that has been identified as causing an allergic reaction. Here, we developed a quantification method of Gly m 5.0101 with multiple reaction monitoring using the synthetic peptide 194NPFLFGSNR202 as the external standard. Firstly, the ground soybean was defatted and extracted with a protein extraction buffer. Then the crude extract was on-filter digested by trypsin and analyzed by liquid chromatography-tandem mass spectrometry. The selected peptide exhibited a detection limit of 0.48 ng/mL and a linear relationship in a concentration range from 1.6 to 500 ng/mL (r² > 0.99). The developed method was successfully applied to quantify the Gly m 5.0101 level in dozens of soybean varieties from different sources and soybean products derived from different processing techniques. The developed method could be used to further analyze ß-conglycinin in soybean seeds combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis.

A ratiometric fluorescence probe for imaging sulfur dioxide derivatives in the mitochondria of living cells.

Although sulfur dioxide (SO2) plays an essential role in several physiological processes, monitoring of intracellular SO2 at subcellular levels remains challenging due to the lack of rapid and sensitive methods for its quantification in a 100% aqueous solution. Herein, a new hemicyanine dyes-based fluorescence probe, NBD-Id, was designed and synthesized for the detection of SO2 derivatives in pure aqueous solution and living cells. By virtue of a specific 1,4-addition reaction of SO32-HSO3- and the polymethine chain of hemicyanine, significant changes in the absorption and fluorescence emission spectra were observed in less than 20 seconds. The ratiometric fluorescence (F467/F593) detection of SO2 derivatives was then obtained with high sensitivity (detection limit 3.6 nM). It was noted that NBD-Id has a specific response towards SO2 derivatives without interference from other anions and biomolecules. Intracellular fluorescence imaging indicated that NBD-Id is cell membrane permeable and mainly distributed within the mitochondria. Therefore, ratiometric fluorescence imaging of SO2 derivatives in the mitochondria of MCF-7 cells was successfully demonstrated.

Synthesis and Application of an Aldazine-Based Fluorescence Chemosensor for the Sequential Detection of Cu²âº and Biological Thiols in Aqueous Solution and Living Cells.

A fluorescence chemosensor, 2-hydroxy-1-naphthaldehyde azine (HNA) was designed and synthesized for sequential detection of Cu(2+) and biothiols. It was found that HNA can specifically bind to Cu(2+) with 1:1 stoichiometry, accompanied with a dramatic fluorescence quenching and a remarkable bathochromic-shift of the absorbance peak in HEPES buffer. The generated HNA-Cu(2+) ensemble displayed a "turn-on" fluorescent response specific for biothiols (Hcy, Cys and GSH) based on the displacement approach, giving a remarkable recovery of fluorescence and UV-Vis spectra. The detection limits of HNA-Cu(2+) to Hcy, Cys and GSH were estimated to be 1.5 µM, 1.0 µM and 0.8 µM, respectively, suggesting that HNA-Cu(2+) is sensitive enough for the determination of thiols in biological systems. The biocompatibility of HNA towards A549 human lung carcinoma cell, was evaluated by an MTT assay. The capability of HNA-Cu(2+) to detect biothiols in live A549 cells was then demonstrated by a microscopy fluorescence imaging assay.

NBD-based fluorescent chemosensor for the selective quantification of copper and sulfide in an aqueous solution and living cells.

Chemosensors play important roles in cation and anion recognition in biological, industrial, and environmental processes. Although many efforts have been made to develop artificial fluorescent receptors for Cu(2+) and S(2-), their applications in the detection in bulk solutions are limited. In this work, we report a novel fluorescence chemosensor (NL) based on the 7-nitrobenz-2-oxa-1,3-diazole (NBD) fluorophore for the quantification of Cu(2+) and S(2-) in single intact cells. NL specifically binds to Cu(2+) in the presence of other competing cations, and evident changes in UV-vis and fluorescence spectra in HEPES buffer are noticed. Based on the displacement approach, the selective sense S(2-) with the in situ generated NL-Cu(2+) ensemble gives a remarkable recovery of fluorescence and UV-vis absorption spectra. The detection limits of NL for Cu(2+) and NL-Cu(2+) for S(2-) were estimated to be 1.6 nM and 0.17 µM, respectively. NL and the resultant complex NL-Cu(2+) exhibit low cytotoxicity and cell-membrane permeability, which makes them capable of Cu(2+) and S(2-) imaging and quantification in living MDA-MB-231 cells.

Bland-Altman agreement analysis between CT predicted and surgical peritoneal cancer index in pseudomyxoma peritonei of appendiceal origin.

Peritoneal cancer index (PCI) is the surgical variable most commonly used to quantify the extent of peritoneal metastases for pseudomyxoma peritonei (PMP) patients. The present study aimed to investigate the agreement between CT predicted and surgical PCI by the Bland-Altman method for PMP of appendiceal origin. A total of 167 PMP patients of appendiceal origin were included between 2016 and 2021. Bland-Altman analysis was performed for both total PCI and selected PCI (regions 2 + 9-12). After the Bland-Altman plot was drawn, the mean bias and its 95% limit of agreements (LoAs) was quantified. Besides, the correlation coefficients between CT-PCI and surgical PCI were also been calculated. The Bland-Altman plot showed the mean bias ± SD between total CT-PCI and surgical PCI as 0.431 ± 3.005, with the LoAs from - 5.459 to 6.321. There were nine points of difference in total PCI exceeded the 95% LoAs, with the rate of 5.39% (9/167). As for selected CT-PCI, Bland-Altman plot showed the mean bias ± SD between selected CT-PCI and surgical PCI as - 0.287 ± 1.955, with the LoAs from - 4.118 to 3.544. There were ten points of difference in selected PCI exceeded the 95% LoAs, with the rate of 5.99% (10/167). The Spearman's rank correlation coefficient between total CT-PCI and surgical PCI was 0.911, P < 0.001, as for selected CT-PCI and surgical PCI, the coefficient was 0.909, P < 0.001. Although there was a strong correlation for both total and selected CT-PCI with surgical PCI, however, the agreement is still not good in Bland-Altman analysis, which suggested that CT-PCI cannot predict surgical PCI accurately even in professional PMP treatment centers. In brief explanation, CT makes it difficult to distinguish the borderline between tumor tissue and mucus and to detect tumor lesions in the small intestine regions, which caused overestimation or underestimation by CT-PCI. In the future, a multiple linear regression model based on CT-PCI might accurately predict surgical PCI preoperatively.

Bioconversion of vitamin D3 to bioactive calcifediol and calcitriol as high-value compounds.

Biological catalysis is an important approach for the production of high-value-added compounds, especially for products with complex structures. Limited by the complex steps of chemical synthesis and low yields, the bioconversion of vitamin D3 (VD3) to calcifediol and calcitriol, which are natural steroid products with high added value and significantly higher biological activity compared to VD3, is probably the most promising strategy for calcifediol and calcitriol production, and can be used as an alternative method for chemical synthesis. The conversion efficiency of VD3 to calcifediol and calcitriol has continued to rise in the past few decades with the help of several different VD3 hydroxylases, mostly cytochrome P450s (CYPs), and newly isolated strains. The production of calcifediol and calcitriol can be systematically increased in different ways. Specific CYPs and steroid C25 dehydrogenase (S25DH), as VD3 hydroxylases, are capable of converting VD3 to calcifediol and calcitriol. Some isolated actinomycetes have also been exploited for fermentative production of calcifediol and calcitriol, although the VD3 hydroxylases of these strains have not been elucidated. With the rapid development of synthetic biology and enzyme engineering, quite a lot of advances in bioproduction of calcifediol and calcitriol has been achieved in recent years. Therefore, here we review the successful strategies of promoting VD3 hydroxylation and provide some perspective on how to further improve the bioconversion of VD3 to calcifediol and calcitriol.

A Novel Fluorescence Probe for the Reversible Detection of Bisulfite and Hydrogen Peroxide Pair in Vitro and in Vivo.

The detection of changes in the reactive oxygen species (ROS)/reactive sulfur species (RSS) couple is important for studying the cellular redox state. Herein, we developed a 1,8-naphthalimide-based fluorescence probe (NI) for the reversible detection of bisulfite (HSO3 - ) and hydrogen peroxide (H2 O2 ) in vitro and in vivo. NI has been designed with a reactive ethylene unit which specifically reacts with HSO3 - by a Michael addition reaction mechanism, resulting in the quenching of yellow fluorescence at 580 nm and the appearing of green fluorescence at 510 nm upon excitation at 500 nm and 430 nm, respectively. The addition product (NI-HSO3 ) could be specifically oxidized to form the original C=C bond of NI, recovering the fluorescence emission and color. The detection limits of NI for HSO3 - and NI-HSO3 for H2 O2 were calculated to be 2.05 µM and 4.23 µM, respectively. The reversible fluorescence response of NI towards HSO3 - /H2 O2 couple can be repeated for at least five times. NI is reliable at a broad pH range (pH 3.0-11.5) and features outstanding selectivity, which enabled its practical applications in biological and food samples. Monitoring the reversible and dynamic inter-conversion between HSO3 - and H2 O2 in vitro and in vivo has been verified by fluorescence imaging in live HeLa cells, adult zebrafish and nude mice. Moreover, NI has been successfully applied to detect of HSO3 - levels in food samples.

Development of a new water-soluble fluorescence probe for hypochlorous acid detection in drinking water.

Responsive small-molecule fluorescence probe specific for target analyte detection is an emerging technology for food safety and quality analysis. In this work, we report a new water soluble small-molecule fluorescence probe (PG) for the detection of hypochlorous acid (HOCl) in drinking water samples. Probe PG was developed by coupling of a glucosamine into 10-methyl-10H-phenothiazine fluorophore with a HOCl-responsive C=N bond. The thioether is another recognition site that can be oxidized to be sulfoxide in water. Due to the specific reactions triggered by HOCl, probe PG's absorption band is blue shifted from 388 to 340 nm, and fluorescence at 488 nm is more than 55-fold enhanced. Probe PG features high fluorescence stability in PBS buffer with varied pH, fast response and high selectivity to HOCl. The application of the probe PG for HOCl detection in real-world samples is demonstrated by HOCl detection in drinking water, including tap water, purified water, and spring water samples. The recoveries of this method for HOCl detection in drinking water are in the range of 99.17-102.3%. This work thus provides a new method for HOCl detection in drinking water with high precision and accuracy.

Effect of grain morphology on the degradation behavior of Mg-4 wt% Zn alloy in Hank's solution.

The degradation behavior of Mg-4 wt% Zn alloy with three different microstructures was examined in Hank's solution at 37 °C by electrochemical measurements and immersion tests in this study. The results show that the sample with cellular structure exhibits a more positive corrosion potential, lower corrosion current density, larger impedance and more protective film than samples with columnar dendritic and equiaxed dendritic structure. The higher corrosion resistance is attributed to the preferred orientation, eliminating susceptible grain boundaries and reduced secondary phases.

Red-Emission Probe for Ratiometric Fluorescent Detection of Bisulfite and Its Application in Live Animals and Food Samples.

Key roles of bisulfite (HSO3 -) in food quality assurance and human health necessitate a reliable analytical method for rapid, sensitive, and selective detection of HSO3 -. Herein, a new red-emitting ratiometric fluorescence probe, BIQ, is reported for sensitive and selective detection of HSO3 - in food samples and live animals. Probe BIQ recognizes HSO3 - via a 1,4-nucleophilic addition reaction. As a result of this specific reaction, emission intensities at 625 and 475 nm are dramatically changed, allowing the detection of HSO3 - in a ratiometric fluorescence model in an aqueous solution. The obvious changes of solution color from pink to transparent and fluorescence color from rose-red to cyan allow the detection of HSO3 - by naked eyes. Furthermore, probe BIQ has fast response in color and fluorescence (<2 min), excellent selectivity, and a low detection limit (0.29 µM), which enables its application in HSO3 - detection in food samples and live organisms. The practical applications of probe BIQ are then demonstrated by the visualization of HSO3 - in live animals (zebrafish and nude mouse) as well as the determination of HSO3 - in white wine and sugar.

Effects of L-lysine·H2SO4 product on the intestinal morphology and liver pathology using broiler model.

BACKGROUND: Lysine is used widely in livestock production due to the shortage of feed protein resources. L-lysine·H2SO4 contains L-lysine sulphate as well as fermentation co-products which contain other amino acids and phosphorus. However, there are few articles about L-lysine·H2SO4 product regarding intestinal morphology and liver pathology of broiler chickens. In this article, we focus on the absorption and metabolism of L-lysine·H2SO4 revealed in the variation of intestinal morphology and liver pathology to determine the tolerance of chicks for L-lysine·H2SO4. METHODS: To evaluate the tolerance of broilers for L-lysine·H2SO4, 240 one day old broilers were allocated randomly to one of five dietary treatments which included corn-soybean diets containing 0, 1%, 4%, 7% or 10% L-lysine·H2SO4 (L-lysine content = 55%). RESULTS: Supplementation of 1% L-lysine·H2SO4 in the diet had no negative effects. However, 4%, 7% or 10% L-lysine·H2SO4 supplementation produced negative responses on broiler performance, carcass characteristics, blood biochemistry, and particularly on intestinal morphology and liver pathology compared with broilers fed the control diet. CONCLUSION: Our results show that supplementation with 1% L-lysine·H2SO4 had no negative effects on performance, carcass characteristics, blood biochemistry, intestinal morphology and liver pathology in broilers, but supplementation with 4%, 7% or 10% L-lysine·H2SO4 produced a negative response, particularly with respect to intestinal morphology and liver pathology.

Rapid Response Fluorescence Probe Enabled In Vivo Diagnosis and Assessing Treatment Response of Hypochlorous Acid-Mediated Rheumatoid Arthritis.

Diagnosis and early assessment of the treatment response of rheumatoid arthritis (RA) necessitates a reliable bioanalytical method for rapid, sensitive, and specific detection of the hypochlorous acid (HOCl) biomarker in inflammatory diseases. Herein, two fluorescence probes, Probe-1 and Probe-2 are developed for quantitative monitoring and visualization of inflammatory response-related HOCl levels in vitro and in vivo. In the presence of HOCl, fluorescence "OFF-ON" response is obtained for both the probes as a result of specific HOCl-triggered C=N bond cleavage reaction. Probe-1 and Probe-2 feature rapid response (<4 s), a high degree of sensitivity and selectivity toward HOCl, which allow them to be used for quantification of HOCl in a simulated physiological condition. Using Probe-2 as the probe, fluorescence imaging and flow cytometry analysis of HOCl levels in lysosome of inflammatory mimic cells, visualization of HOCl generation in endotoxin-induced inflammation of adult zebrafish and RA of mice are possible. Probe-2 exhibits high effectiveness for early assessment of the treatment response of HOCl-mediated RA in mice with an antiarthritic drug, methotrexate (MTX). The results demonstrate that Probe-2 is a powerful tool for future studies on diagnosis and monitoring treatment efficiency in a broad range of inflammatory diseases, including RA.

Numerical simulation of non-dendritic structure formation in Mg-Al alloy solidified with ultrasonic field.

The formation of non-dendritic structure of Mg alloy solidified with ultrasonic treatment was investigated by numerical simulation and experiment. The models of nucleation and crystal growth involved the effects of ultrasonic cavitation and acoustic streaming were built. Based on the models, the grain refinement and the microstructure change from dendrite to non-dendritic structure of a Mg-Al alloy were numerically simulated by cellular automata method. The simulation and experimental results indicated that the ultrasonic cavitation strongly contributes to the grain refinement by improving nucleation, while the acoustic streaming is mainly responsible for the formation of non-dendritic structure.

A highly sensitive electrochemical biosensor for phenol derivatives using a graphene oxide-modified tyrosinase electrode.

The fabrication, characterization and analytical performance were investigated for a phenol biosensor based on the covalent bonding of tyrosinase (TYR) onto a graphene oxide (GO)-modified glassy carbon electrode (GCE) via glutaraldehyde (GA). The surface morphology of the modified electrode was studied by atomic force microscope (AFM) and field-emission scanning electron microscopy (FE-SEM). The fabricated TYR/GA/GO/GCE biosensor showed very good stability, reproducibility, sensitivity and practical usage. The catechol biosensor exhibited a wide sensing linear range from 5×10-8M to 5×10-5M, a lower detection limit of 3×10-8M, a current maximum (Imax) of 65.8µA and an apparent Michaelis constant (Kmapp) of 169.9µM.

Responsive Fluorescence Probe for Selective and Sensitive Detection of Hypochlorous Acid in Live Cells and Animals.

The development of effective bioanalytical methods for rapid, sensitive and specific detection of HOCl in vitro and in vivo plays a key role for better understanding the roles of this molecule in normal and diseased conditions, but remains challenging due to the highly reactive nature of HOCl and the complicated biological conditions. In this work, a new fluorescence probe, PQI, was developed for monitoring of the HOCl level in biological samples. PQI was easily synthesized by a one-step condensation reaction. Upon addition of HOCl, significant changes in the absorption spectra and the color of the solution were noticed, facilitating the "naked eye" detection of HOCl in PBS buffer. The fluorescence of PQI was found to be significantly increased within a few seconds, leading to "OFF-ON" fluorescence response towards HOCl. The sensing mechanism, oxidation of thioether by HOCl, was confirmed by HRMS titration analysis. PQI features a large Stokes shift, high sensitivity and selectivity, and rapid fluorescence response towards HOCl. Quantitative detection of HOCl in single live cells was demonstrated through fluorescence imaging and flow cytometry analysis. PQI was then successfully used in visualisation of HOCl in live zebrafish and nude mice.

Valine Supplementation in a Reduced Protein Diet Regulates Growth Performance Partially through Modulation of Plasma Amino Acids Profile, Metabolic Responses, Endocrine, and Neural Factors in Piglets.

The objective of this study was to investigate whether valine (Val) supplementation in a reduced protein (RP) diet regulates growth performance associated with the changes in plasma amino acids (AAs) profile, metabolism, endocrine, and neural system in piglets. Piglets or piglets with a catheter in the precaval vein were randomly assigned to two treatments, including two RP diets with standardized ileal digestible (SID) Val:Lysine (Lys) ratio of 0.45 and 0.65, respectively. The results indicated that piglets in the higher Val:Lys ratio treatment had higher average daily feed intake (ADFI) ( P < 0.001), average daily gain (ADG) ( P = 0.001), feed conversion ratio (FCR) ( P = 0.004), lower plasma urea nitrogen ( P = 0.032), expression of gastric cholecystokinin (CCK), and hypothalamic pro-opiomelanocortin (POMC). Plasma AAs profiles including postprandial plasma essential AAs (EAAs) profile and in serum, muscle, and liver involved in metabolism of AAs and fatty acids were significantly different between two treatments. In conclusion, Val influenced growth performance associated with metabolism of AAs and fatty acids and both endocrine and neural system in piglets.

Lactobacillus reuteri I5007 Modulates Intestinal Host Defense Peptide Expression in the Model of IPEC-J2 Cells and Neonatal Piglets.

Modulation of the synthesis of endogenous host defense peptides (HDPs) by probiotics represents a novel antimicrobial approach for disease control and prevention, particularly against antibiotic-resistant infections in human and animals. However, the extent of HDP modulation by probiotics is species dependent and strain specific. In the present study, The porcine small intestinal epithelial cell line (IPEC-J2) cells and neonatal piglets were used as in-vitro and in-vivo models to test whether Lactobacillus reuteri I5007 could modulate intestinal HDP expression. Gene expressions of HDPs, toll-like receptors, and fatty acid receptors were determined, as well as colonic short chain fatty acid concentrations and microbiota. Exposure to 108 colony forming units (CFU)/mL of L. reuteri I5007 for 6 h significantly increased the expression of porcine ß-Defensin2 (PBD2), pBD3, pBD114, pBD129, and protegrins (PG) 1-5 in IPEC-J2 cells. Similarly, L. reuteri I5007 administration significantly increased the expression of jejunal pBD2 as well as colonic pBD2, pBD3, pBD114, and pBD129 in neonatal piglets (p < 0.05). This was probably associated with the increase in colonic butyric acid concentration and up-regulating expression of Peroxisome Proliferator-Activated Receptor-γ (PPAR-γ) and G Protein-Coupled Receptor 41 (GPR41) (p < 0.05), but not with stimulation of Pattern-Recognition Receptors. Additionally, supplementation with L. reuteri I5007 in the piglets did not affect the colonic microbiota structure. Our findings suggested that L. reuteri I5007 could modulate intestinal HDP expression and improve the gut health of neonatal piglets, probably through the increase in colonic butyric acid concentration and the up-regulation of the downstream molecules of butyric acid, PPAR-γ and GPR41, but not through modifying gut microbiota structure.