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Talanta Dec 2023Superoxide anion (O) is typically produced in living cells and organisms, while excess O may cause unexpected damage, so monitoring and scavenging the O is of...
Superoxide anion (O) is typically produced in living cells and organisms, while excess O may cause unexpected damage, so monitoring and scavenging the O is of considerable significance to exploring physiological and pathological process. In this study, a Cu-based metal-organic framework (Cu-MOF) which comprise sequential Cu metal ion and conductive organic 2,5-dicarboxylic acid-3,4-ethylene dioxythiophene is synthesized to mimic superoxide dismutase (SOD), in which Cu is the essence of active site. On one hand, the Cu-MOF possesses excellent electrocatalytic activity to detect O at -0.05 V, biased at which potential the electrode showed good linearity toward O with detection limit of 0.283 μM and interference immunity for AA, DA, UA, 5-HT and HO. The Cu-MOF modified microelectrode was applied for measuring the O released from living cells real time and monitoring O generation in rat brain. On the other hand, this Cu-MOF has the catalytic activity to mimic the superoxide dismutase for scavenging O in HeLa cells effectively. This work provides a methodology to design metal ion based enzyme mimetic for analyzing and scavenging O in cells and in vivo.
Topics: Humans; Animals; Rats; Superoxides; Superoxide Dismutase; Metal-Organic Frameworks; HeLa Cells; Hydrogen Peroxide; Microelectrodes
PubMed: 37429254
DOI: 10.1016/j.talanta.2023.124860 -
Journal of Visualized Experiments : JoVE Mar 2024Reactive oxygen species (ROS) are highly unstable oxygen-containing molecules. Their chemical instability makes them extremely reactive and gives them the ability to...
Reactive oxygen species (ROS) are highly unstable oxygen-containing molecules. Their chemical instability makes them extremely reactive and gives them the ability to react with important biological molecules such as proteins, nucleic acids, and lipids. Superoxide anions are important ROS generated by the reduction of molecular oxygen reduction (i.e., acquisition of one electron). Despite their initial implication exclusively in aging, degenerative, and pathogenic processes, their participation in important physiological responses has recently become apparent. In the vascular system, superoxide anions have been shown to modulate the differentiation and function of vascular smooth muscle cells, the proliferation and migration of vascular endothelial cells in angiogenesis, the immune response, and the activation of platelets in hemostasis. The role of superoxide anions is particularly important in the dysregulation of platelets and the cardiovascular complications associated with a plethora of conditions, including cancer, infection, inflammation, diabetes, and obesity. It has, therefore, become extremely relevant in cardiovascular research to be able to effectively measure the generation of superoxide anions by human platelets, understand the redox-dependent mechanisms regulating the balance between hemostasis and thrombosis and, eventually, identify novel pharmacological tools for the modulation of platelet responses leading to thrombosis and cardiovascular complications. This study presents three experimental protocols successfully adopted for the detection of superoxide anions in platelets and the study of the redox-dependent mechanisms regulating hemostasis and thrombosis: 1) dihydroethidium (DHE)-based superoxide anion detection by flow cytometry; 2) DHE-based superoxide anion visualization and analysis by single platelet imaging; and 3) spin probe-based quantification of superoxide anion output in platelets by electron paramagnetic resonance (EPR).
Topics: Humans; Superoxides; Reactive Oxygen Species; Endothelial Cells; Oxygen; Thrombosis
PubMed: 38619265
DOI: 10.3791/66647 -
Water Research Sep 2023Traditional advanced oxidation processes suffer from low availability of ultrashort lifetime radicals and declining stability of catalysts. Co nanoparticles in hollow...
Traditional advanced oxidation processes suffer from low availability of ultrashort lifetime radicals and declining stability of catalysts. Co nanoparticles in hollow bimetallic metal-organic frameworks (Co@MOFs) were synthesized via a solvothermal method. Nanoconfinement and peroxymonocarbonate (PMC) degradation system endows Co@MOFs with high catalytic activity and stability even in the actual water matrices. The nanocomposites exhibited 100-200 nm polyhedron structure with irregular nanocavity between the 20 nm shell and multicores. Co nanoparticles were completely encapsulated by the Fe-MOF-5 shell according to the X-ray diffraction and photoelectron spectra. Both 0.8 nm micropores and 3.6 nm mesopores were proven to be present. The yolk-shell Co@MOFs exhibited higher catalytic performance than that of Co nanoparticles, hollow Fe-MOF-5 and its core-shell counterpart toward PMC activation during sulfamethoxazole degradation. The catalytic activities of Co@MOFs for the activation of unsymmetrical peroxides (PMC and peroxymonosulfate) were much higher than those for the symmetrical peroxides (HO and persulfate) and the heterogeneous catalysis was dominant in the Co@MOFs activated HO and PMC systems. The MOF stability was the highest and metal leakages were the least in the activated PMC system among the four peroxides because of mild reaction conditions and the alkalescent solution (pH = 8.3-8.4). Furthermore, the high removal efficiencies (>94%) and degradation rates could be maintained in the different actual water matrices due to the confinement effects. The contributions of carbonate and hydroxyl radicals were primary for sulfamethoxazole degradation, and superoxide anion and singlet oxygen also played essential roles according to scavenging experiments and time-series spin-trapping electron spin resonance spectra. Six degradation pathways were proposed according to 26 intermediate identification and the pharmacophores of more than 80% intermediates were destroyed, which would benefit subsequent biological treatment. Successful combination of nanoconfinement and PMC might provide a new effective solution for pollution remediation.
Topics: Ferric Compounds; Hydrogen Peroxide; Metal-Organic Frameworks; Carbonates; Nanoparticles; Peroxides; Anti-Bacterial Agents
PubMed: 37480599
DOI: 10.1016/j.watres.2023.120340 -
Analytical and Bioanalytical Chemistry Jun 2024Nucleotide sugars (NS) fulfil important roles in all living organisms and in humans, related defects result in severe clinical syndromes. NS can be seen as the...
Nucleotide sugars (NS) fulfil important roles in all living organisms and in humans, related defects result in severe clinical syndromes. NS can be seen as the "activated" sugars used for biosynthesis of a wide range of glycoconjugates and serve as substrates themselves for the synthesis of other nucleotide sugars. NS analysis is complicated by the presence of multiple stereoisomers without diagnostic transition ions, therefore requiring separation by liquid chromatography. In this paper, we explored weak anion-exchange/reversed-phase chromatography on a hybrid column for the separation of 17 nucleotide sugars that can occur in humans. A robust and reproducible method was established with intra- and inter-day coefficients of variation below 10% and a linear range spanning three orders of magnitude. Application to patient fibroblasts with genetic defects in mannose-1-phosphate guanylyltransferase beta, CDP-L-ribitol pyrophosphorylase A, and UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase showed abnormal levels of guanosine-5'-diphosphate-α-D-mannose (GDP-Man), cytidine-5'-diphosphate-L-ribitol (CDP-ribitol), and cytidine-5'-monophosphate-N-acetyl-β-D-neuraminic acid (CMP-Neu5Ac), respectively, in consonance with expectations based on the diagnosis. In conclusion, a novel, semi-quantitative method was established for the analysis of nucleotide sugars that can be applied to diagnose several genetic glycosylation disorders in fibroblasts and beyond.
Topics: Humans; Fibroblasts; Tandem Mass Spectrometry; Chromatography, Ion Exchange; Chromatography, Reverse-Phase; Nucleotides; Anions; Liquid Chromatography-Mass Spectrometry
PubMed: 38676823
DOI: 10.1007/s00216-024-05313-w -
Methods in Enzymology 2024There is intense interest in removing fluorinated compounds from the environment, environments are most efficiently remediated by microbial enzymes, and defluorinating...
There is intense interest in removing fluorinated compounds from the environment, environments are most efficiently remediated by microbial enzymes, and defluorinating enzymes are readily monitored by fluoride determination. Fluorine is the most electronegative element. Consequently, all mechanisms of enzymatic C-F bond cleavage produce fluoride anion, F. Therefore, methods for the determination of fluoride are critical for C-F enzymology and apply to any fluorinated organic compounds, including PFAS, or per- and polyfluorinated alkyl substances. The biodegradation of most PFAS chemicals is rare or unknown. Accordingly, identifying new enzymes, or re-engineering the known defluorinases, will require rapid and sensitive methods for measuring fluoride in aqueous media. Most studies currently use ion chromatography or fluoride specific electrodes which are relatively sensitive but low throughput. The methods here describe refashioning a drinking water test to efficiently determine fluoride in enzyme and cell culture reaction mixtures. The method is based on lanthanum alizarin complexone binding of fluoride. Reworking the method to a microtiter well plate format allows detection of as little as 4 nmol of fluoride in 200 μL of assay buffer. The method is amenable to color imaging, spectrophotometric plate reading and automated liquid handling to expedite assays with thousands of enzymes and/or substrates for discovering and improving enzymatic defluorination.
Topics: Fluorides; Drinking Water; Halogenation; Enzyme Assays
PubMed: 38658089
DOI: 10.1016/bs.mie.2023.12.020 -
ACS Applied Materials & Interfaces Mar 2024The unique structural sensitivity of photonic crystals (PCs) endows them with stretchable or elastic tunability for light propagation and spontaneous emission...
The unique structural sensitivity of photonic crystals (PCs) endows them with stretchable or elastic tunability for light propagation and spontaneous emission modulation. Hydrogel PCs have been demonstrated to have biocompatibility and flexibility for potential human health detection and environmental security monitoring. However, current elastic PCs still possess a fixed elastic modulus and uncontrollable structural colors based on a tunable elastic modulus, posing considerable challenges for in situ detection, particularly in wearable or portable sensing devices. In this work, we introduced a novel chemo-mechanical transduction mechanism embedded within a photonic crystal nanomatrix, leading to the creation of structural colors and giving rise to a visual gustation sensing experience. By utilizing the captivating structural colors generated by the hydrogel PC, we employ abundant optical information to identify various analytes. The finite element analysis proved the electric field distribution in the PC matrix during stretch operations. The elastic-optical behaviors with various chemical cosolvents, including cations, anions, saccharides, or organic acids, were investigated. The mechanism of the Hofmeister effect regulating the elasticity of hydrogels was demonstrated with the network nanostructure of the hydrogels. The hydrogel PC matrix demonstrates remarkable capability in efficiently distinguishing a wide range of cations, anions, saccharides, and organic acids across various concentrations, mixtures, and even real food samples, such as tastes and soups. Through comprehensive research, a precise relationship between the structural colors and the elastic modulus of hydrogel PCs has been established, contributing to the biomatching elastic-optics platform for wearable devices, a dynamic environment, and clinical or health monitoring auxiliary.
Topics: Humans; Taste; Elastic Modulus; Hydrogels; Anions; Cations
PubMed: 38447141
DOI: 10.1021/acsami.3c18892 -
The Journal of Physical Chemistry. A Dec 2023is a marine worm, commonly known as a fireworm, that exhibits bluish-green bioluminescence (BL). The luciferin () and oxyluciferin () during fireworm BL have been...
is a marine worm, commonly known as a fireworm, that exhibits bluish-green bioluminescence (BL). The luciferin () and oxyluciferin () during fireworm BL have been experimentally identified in vitro. The and are the respective starting point and ending point of a series of complicated chemical reactions in the BL. However, the chemical mechanism of the fireworm BL remains largely unknown. Before the experiments provided strong evidence for the mechanism, based on our previously successful studies on several bioluminescent systems, we theoretically proposed the chemical mechanism of the fireworm BL in this article. By means of the spin-flip and time-dependent density functional calculations, we clearly described the complete process from to : under the catalysis of luciferase, undergoes deprotonation and reacts with O to form a dioxetanone anion via the single-electron transfer mechanism; the dioxetanone anion decomposes into the at the first singlet excited state (S) by the gradually reversible charge-transfer-induced luminescence mechanism; and the S- emits light and deexcites to in the ground state.
Topics: Luciferases; Electron Transport; Luminescence; Anions; Luminescent Measurements
PubMed: 38103213
DOI: 10.1021/acs.jpca.3c07409 -
European Journal of Medicinal Chemistry Dec 2023Neutrophils are the most abundant immune cells. However, neutrophil dysregulation leads to acute and chronic inflammation and is involved in various diseases. The aim of...
Neutrophils are the most abundant immune cells. However, neutrophil dysregulation leads to acute and chronic inflammation and is involved in various diseases. The aim of this study was to develop anti-inflammatory agents in human neutrophils. A drug screening was conducted on in-house compounds with the potential to inhibit the respiratory burst, which involves the generation of superoxide anions in human neutrophils. Bioisosteric replacement was then applied to design more active derivatives. The most potent inhibitors of superoxide anion generation activity were compounds 58 and 59, which had IC values of 13.30 and 9.06 nM, respectively. The inhibitory effects of 58 and 59 were reversed by H89, a PKA inhibitor. PDE selective screening indicated that the best inhibitory effects were PDE4B1 and PDE4D2, and the inhibitory activities were 83% and 85%, respectively, at a 10 μM concentration of 59. The final molecular simulation experiment highlighted the slightly different binding poses of 58 and 59 in the PDE4 active site. An in vivo pharmacokinetic study revealed that the half-life of 59 was approximately 79 min when using intravenous bolus administration. This work introduced a new class structure of PDE4 inhibitors resulting in potent neutrophil inactivation activity, with the aim of contributing to new anti-inflammatory drug discovery.
Topics: Humans; Superoxides; Anti-Inflammatory Agents; Phosphodiesterase 4 Inhibitors; Inflammation; Pyrazoles; Neutrophils
PubMed: 37918036
DOI: 10.1016/j.ejmech.2023.115874 -
Journal of Intensive Care Medicine Oct 2023In lactic acidosis, lactate can only explain 30% of the variance in the anion gap (AG), and the elevated AG not explained by lactate is due to unmeasured organic anions...
In lactic acidosis, lactate can only explain 30% of the variance in the anion gap (AG), and the elevated AG not explained by lactate is due to unmeasured organic anions (UOAs). Some studies using less precise surrogates for UOA have suggested that UOA may predict clinical outcomes better than lactate. The aim of this study was to determine whether UOA predicts clinical outcomes better than lactate levels. This was a retrospective cohort study of adult ICU patients with sepsis. Baseline AG and albumin measurements were obtained. An albumin-corrected delta AG was calculated. UOAs were estimated using the formula: Delta AG - serum lactate. A multivariate logistic regression model with its respective ROC curve was constructed to explore the relationship between in-hospital mortality, UOA, and lactate. 526 patients were included. In the combined model examining both lactate and UOA, the odds ratio (OR) [95% CI] for predicting ICU length of stay (LOS) was 1.050 [1.029-1.072] and 1.022 [1.009-1.035], respectively; the OR [95% CI] for predicting in-hospital mortality was 1.224 [1.104-1.358] and 0.997 [0.943-1.054], respectively. The ROC curve for in-hospital mortality demonstrated that the Area Under the Curve (AUC) for lactate, UOA, and combined lactate and UOA was 0.7726, 0.7486, and 0.7732, respectively. The AUC for combined lactate and UOA were not statistically significantly higher than the AUC for lactate alone ( .9193). As expected, serum lactate predicted both ICU LOS and in-hospital mortality. UOA did predict ICU LOS, although the reason for this association is not known. UOA did not predict in-hospital mortality based on the OR and the ROC curve's AUC, contrary to some previous studies. However, our study used a more precise quantitative estimate of UOA, including the use of baseline albumin-corrected AG. Prior studies attempting to identify UOA have identified Krebs cycle intermediates including citrate and isocitrate, suggesting that in our study these anions associated with the Krebs cycle contributed to the UOA.
Topics: Adult; Humans; Acidosis, Lactic; Retrospective Studies; Anions; Lactic Acid; Albumins; Sepsis; ROC Curve
PubMed: 37264611
DOI: 10.1177/08850666231177602 -
Physiologia Plantarum 2023All land-plant cell walls possess hemicelluloses, cellulose and anionic pectin. The walls of their cousins, the charophytic algae, exhibit some similarities to land...
All land-plant cell walls possess hemicelluloses, cellulose and anionic pectin. The walls of their cousins, the charophytic algae, exhibit some similarities to land plants' but also major differences. Charophyte 'pectins' are extractable by conventional land-plant methods, although they differ significantly in composition. Here, we explore 'pectins' of an early-diverging charophyte, Chlorokybus atmophyticus, characterising the anionic polysaccharides that may be comparable to 'pectins' in other streptophytes. Chlorokybus 'pectin' was anionic and upon acid hydrolysis gave GlcA, GalA and sulphate, plus neutral sugars (Ara≈Glc>Gal>Xyl); Rha was undetectable. Most Gal was the l-enantiomer. A relatively acid-resistant disaccharide was characterised as β-d-GlcA-(1→4)-l-Gal. Two Chlorokybus 'pectin' fractions, separable by anion-exchange chromatography, had similar sugar compositions but different sulphate-ester contents. No sugars were released from Chlorokybus 'pectin' by several endo-hydrolases [(1,5)-α-l-arabinanase, (1,4)-β-d-galactanase, (1,4)-β-d-xylanase, endo-polygalacturonase] and exo-hydrolases [α- and β-d-galactosidases, α-(1,6)-d-xylosidase]. 'Driselase', which hydrolyses most land-plant cell wall polysaccharides to mono- and disaccharides, released no sugars except traces of starch-derived Glc. Thus, the Ara, Gal, Xyl and GalA of Chlorokybus 'pectin' were not non-reducing termini with configurations familiar from land-plant polysaccharides (α-l-Araf, α- and β-d-Galp, α- and β-d-Xylp and α-d-GalpA), nor mid-chain residues of α-(1→5)-l-arabinan, β-(1→4)-d-galactan, β-(1→4)-d-xylan or α-(1→4)-d-galacturonan. In conclusion, Chlorokybus possesses anionic 'pectic' polysaccharides, possibly fulfilling pectic roles but differing fundamentally from land-plant pectin. Thus, the evolution of land-plant pectin since the last common ancestor of Chlorokybus and land plants is a long and meandering path involving loss of sulphate, most l-Gal and most d-GlcA; re-configuration of Ara, Xyl and GalA; and gain of Rha.
Topics: Polysaccharides; Pectins; Plants; Polygalacturonase; Embryophyta; Sulfates
PubMed: 38148229
DOI: 10.1111/ppl.14079