RESUMEN
Palladium ion (Pd2+) plays an important role in our daily life, but poses a great threat to the environment and human health. Thus, it is desirable to exploit a rapid and sensitive approach to realize the detection of Pd2+. In this study, a cellulose acetate-based macromolecular fluorescent probe CA-NA-PA was successfully prepared for tracking amounts of Pd2+. CA-NA-PA showed an obvious "on-off" fluorescence response to Pd2+, accompanied by the fluorescence color changed from bright yellow to colorless. CA-NA-PA had some outstanding detection performances such as low detection limit (26 nM), extremely short response time (1 min), good selectivity and anti-interference ability. Based on the advantages of probe mentioned above, CA-NA-PA could realize recognition of Pd2+ concentration in environmental water and soil samples. What's more, the probe CA-NA-PA was applied to image Pd2+ in zebrafish as well as in live onion tissue due to the good biocompatibility and cell membrane permeability of cellulose, suggesting its wide application prospect in biosystems.
RESUMEN
Acute liver injury (ALI) is a frequent and devastating liver disease that has been made more prevalent by the excessive use of chemicals, drugs, and alcohol in modern life. Hypochlorous acid (HClO), an important biomarker of oxidative stress originating mainly from the mitochondria, has been shown to be intimately connected to the development and course of ALI. Herein, a novel BODIPY-based NIR ratiometric fluorescent probe Mito-BS was constructed for the specific recognition of mitochondrial HClO. The probe Mito-BS can rapidly respond to HClO within 20 s with a ratiometric fluorescence response (from 680 nm to 645 nm), 24-fold fluorescence intensity ratio enhancement (I645/I680), a wide pH adaptation range (5-9) and the low detection limit (31 nM). The probe Mito-BS has been effectively applied to visualize endogenous and exogenous HClO fluctuations in living zebrafish and cells based on its low cytotoxicity and prominent mitochondria-targeting ability. Furthermore, the fluorescent probe Mito-BS makes it possible to achieve the non-invasive in-situ diagnosis of ALI through in mice, and provides a feasible strategy for early diagnosis and drug therapy of ALI and its complications.
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Colorantes Fluorescentes , Ácido Hipocloroso , Mitocondrias , Pez Cebra , Ácido Hipocloroso/análisis , Ácido Hipocloroso/metabolismo , Animales , Colorantes Fluorescentes/química , Colorantes Fluorescentes/síntesis química , Ratones , Mitocondrias/metabolismo , Modelos Animales de Enfermedad , Humanos , Compuestos de Boro/química , Imagen Óptica , Enfermedad Hepática Inducida por Sustancias y Drogas/metabolismo , Enfermedad Hepática Inducida por Sustancias y Drogas/diagnóstico por imagenRESUMEN
Detection of highly toxic mercury ions (Hg2+) in actual environmental and biological samples is of significant importance for protecting environment and human health. In this paper, a new ratiometric fluorescent probe BTIA was designed and synthesized from 3-pinone based on Internal Charge Transfer (ICT) mechanism. BTIA could selectively recognize Hg2+ over other competitive analytes with short reaction time (5 s), distinct ratiometric response, strong anti-interference ability, large Stokes shift (200 nm), and low detection limit (2.36 × 10-7 M). Furthermore, BTIA was applicable for detecting Hg2+ in actual water samples and it also performed an excellent imaging capability in living RAW264.7 cells, zebrafish and onion tissue.
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Colorantes Fluorescentes , Límite de Detección , Mercurio , Espectrometría de Fluorescencia , Contaminantes Químicos del Agua , Pez Cebra , Animales , Colorantes Fluorescentes/química , Colorantes Fluorescentes/síntesis química , Mercurio/análisis , Ratones , Células RAW 264.7 , Contaminantes Químicos del Agua/análisis , Cebollas/química , Agua/químicaRESUMEN
Mercury ion (Hg2+) is one of harmful heavy metal ions that can accumulate inside the human organism and cause some health problems. In the article, a highly effective fluorescent probe named EC-T-PCBM was prepared by grafting flavonol derivatives onto ethyl cellulose for the specific recognition of Hg2+. EC-T-PCBM exhibited a remarkable fluorescence light-up response toward Hg2+ with excellent sensitivity. EC-T-PCBM possessed several prominent sensing properties for Hg2+, such as low detection limit (43.9 nM), short response time (5 min), and wide detection pH range (6-9). The response mechanism of EC-T-PCBM to Hg2+ has been verified through 1H NMR titration and DFT computation. Additionally, EC-T-PCBM not only can be used for accurately determining trace amount of Hg2+ in actual environmental water samples, but also can serve as a portable and rapid device by loading it on test strips for sensitive and selective visualization of Hg2+. More importantly, the confocal fluorescence imaging of onion cells suggested the favorable cell membrane permeability of EC-T-PCBM and its prominent ability to continuously monitor the enrichment from Hg2+ within fresh plant tissues.
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Celulosa , Flavonoles , Colorantes Fluorescentes , Mercurio , Mercurio/análisis , Celulosa/química , Celulosa/análogos & derivados , Colorantes Fluorescentes/química , Flavonoles/química , Flavonoles/análisis , Espectrometría de Fluorescencia/métodos , Límite de Detección , Concentración de Iones de Hidrógeno , Contaminantes Químicos del Agua/análisis , Cebollas/química , Imagen Óptica/métodosRESUMEN
Pesticides and microplastics are common pollutants in soil environments, adversely affecting soil organisms. However, the combined toxicological effects of aged microplastics and pesticides on soil organisms are still unclear. In this study, we systematically studied the toxicological effects of azoxystrobin and four different aged polyethylene (PE) microplastics on earthworms (Eisenia fetida). The purpose was to evaluate the effects of aging microplastics on the toxicity of microplastics-pesticides combinations on earthworms. The results showed that different-aged PE microplastics promoted azoxystrobin accumulation in earthworms. Meanwhile, combined exposure to azoxystrobin and aged PE microplastics decreased the body weight of earthworms. Besides, both single and combined exposure to azoxystrobin and aged PE microplastics could lead to oxidative damage in earthworms. Further studies revealed that azoxystrobin and aged PE microplastics damage the intestinal structure and function of earthworms. Additionally, the combination of different aged PE microplastics and azoxystrobin was more toxic on earthworms than single exposures. The PE microplastics subjected to mechanical wear, ultraviolet radiation, and acid aging exhibited the strongest toxicity enhancement effects on earthworms. This high toxicity may be related to the modification of PE microplastics caused by aging. In summary, these results demonstrated the enhancing effects of aged PE microplastics on the toxicity of pesticides to earthworms. More importantly, aged PE microplastics exhibited stronger toxicity-enhancing effects in the early exposure stages. This study provides important data supporting the impact of different aged PE microplastics on the environmental risks of pesticides.
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Oligoquetos , Plaguicidas , Pirimidinas , Contaminantes del Suelo , Estrobilurinas , Animales , Microplásticos/toxicidad , Plásticos/toxicidad , Polietileno/toxicidad , Rayos Ultravioleta , Contaminantes del Suelo/análisis , Estrés Oxidativo , Suelo/químicaRESUMEN
Plastic film mulching can maintain soil water and heat conditions, promote plant growth and thus generate considerable economic benefits in agriculture. However, as they age, these plastics degrade and form microplastics (MPs). Additionally, pesticides are widely utilized to control organisms that harm plants, and they can ultimately enter and remain in the environment after use. Pesticides can also be sorbed by MPs, and the sorption kinetics and isotherms explain the three stages of pesticide sorption: rapid sorption, slow sorption and sorption equilibrium. In this process, hydrophobic and partition interactions, electrostatic interactions and valence bond interactions are the main sorption mechanisms. Additionally, small MPs, biodegradable MPs and aged conventional MPs often exhibit stronger pesticide sorption capacity. As environmental conditions change, especially in simulated biological media, pesticides can desorb from MPs. The utilization of pesticides by environmental microorganisms is the main factor controlling the degradation rate of pesticides in the presence of MPs. Pesticide sorption by MPs and size effects of MPs on pesticides are related to the internal exposure level of biological pesticides and changes in pesticide toxicity in the presence of MPs. Most studies have suggested that MPs exacerbate the toxicological effects of pesticides on sentinel species. Hence, the environmental risks of pesticides are altered by MPs and the carrier function of MPs. Based on this, research on the affinity between MPs and various pesticides should be systematically conducted. During agricultural production, pesticides should be cautiously selected and used plastic film to ensure human health and ecological security.
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Microplásticos , Plaguicidas , Humanos , Anciano , Microplásticos/toxicidad , Microplásticos/química , Plásticos/química , Plaguicidas/toxicidad , Plaguicidas/química , Agricultura , Suelo , AdsorciónRESUMEN
Toxic mercury ions (Hg2+) can cause serious environmental pollution and accumulate in living organisms via the food chain. Therefore, monitoring Hg2+ is crucial in ensuring the safety of ecosystems and organisms. In this work, a novel ratiometric fluorescent probe CMT (5-(4-(diphenylamino)phenyl)-1-(7-hydroxy-coumarin-3-yl)-4-pentene-1,3-dione) based on coumarin was developed for detecting Hg2+, which displayed obvious fluorescence changes, a low detection limit (2.24 × 10-7 M), good selectivity, and a large Stokes shift (255 nm). The CMT probe could detect Hg2+ in real environmental soil and water samples. Furthermore, the CMT probe enabled the naked-eye detection of Hg2+ using test paper experiments. CMT was also applied for fluorescence imaging in living zebrafish and plants. This work provides a highly efficient tool for monitoring Hg2+ in environmental samples and biological systems.
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Colorantes Fluorescentes , Mercurio , Animales , Pez Cebra , Ecosistema , Mercurio/análisis , CumarinasRESUMEN
Fluorescent sensors available for metal ions detection have been extensively developed in recent years. However, developing an ultrasensitive fluorescent sensor for highly selectively detecting Al3+ based on cellulose remains a challenge. In this study, an ethylcellulose-based flavonol fluorescent sensor named EC-BHA was synthesized by the esterification of ethylcellulose (EC) with a new flavonol derivative 4-(2-(2,3-bis(ethoxymeothy)phenyl)-3-hydroxy-4-oxo-4-H-chromen-7-yl) benzoic acid (BHA). The fluorescence intensity of EC-BHA exhibited a 180-fold increase at 490 nm after binding with Al3+ and provided an ultralow detection limit of 13.0 nM. The sensor showed some exceptional sensing properties including a broad pH range (4-10), large Stokes shifts (190 nm), and a short response time (3 min). This sensor was successfully applied for determining trace Al3+ in food samples as well as in plant tissue. Moreover, the electrostatic spun film EBP was fabricated by blending EC-BHA with PS (polystyrene) via electrostatic spinning technique and utilized for selective detection of Al3+ as soon as possible.
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Aluminio , Colorantes Fluorescentes , Colorantes Fluorescentes/química , Aluminio/química , Celulosa/química , FlavonolesRESUMEN
Nanotechnology has been widely applied for pesticide carriers, which is an important way to improve the utilization, stability, and sustained release of pesticides. Mesoporous silica nanoparticles (MSNs) are a nanomaterial with adjustable particle and pore sizes, with a high specific surface area and good biocompatibility. Rotenone is a non-systemic botanical insecticide that is easily degraded in the environment. We used a modified soft-template method to prepare MSNs, in which rotenone was loaded using the solvent evaporation method. The prepared rotenone nanopesticide based on mesoporous silica showed considerable drug loading rates of 33.2%. Moreover, the prepared rotenone nanoparticles showed improved photostability and sustained release behavior, which improved the translocation of rotenone in tomato plants. Finally, the rotenone nanoparticles displayed superior insecticidal activity compared to traditional preparations. In summary, the rotenone nanopesticide improved the persistence and utilization rates of rotenone. These findings are of significance in reducing pesticide usage, mitigating environmental pollution, and ensuring food safety.
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Insecticidas , Nanopartículas , Plaguicidas , Portadores de Fármacos , Rotenona , Preparaciones de Acción Retardada , Dióxido de Silicio , Plaguicidas/metabolismo , PorosidadRESUMEN
Al3+ is commonly utilized in daily life, however, the excessive accumulation of Al3+ within organisms can result in severe health problems. Herein, a highly efficient fluorescent probe EC-HTC for Al3+ was synthesized through chemical modification of ethyl cellulose. This probe exhibited a significant fluorescence enhancement response to Al3+, and it interestingly also possessed an obvious aggregation-induced emission (AIE) effect. The detection limit of probe EC-HTC for Al3+ was as low as 0.23 µM, and its pH usage range was as wide as 5-10. The complexation ratio of EC-HTC with Al3+ was determined to be 1:1 based on Job's plot, which was further confirmed by 1H NMR titration and HRMS analysis. Moreover, the probe EC-HTC was successfully employed for the determination of Al3+ in environmental and food samples. In addition, the probe EC-HTC compositing PS (polystyrene) electrostatic spun fiber membranes EHP with high specific surface area were prepared to achieve the rapid and portable detection of Al3+.
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Chalcona , Chalconas , Colorantes Fluorescentes , Celulosa , Espectrometría de FluorescenciaRESUMEN
Pesticides inevitably enter aquatic environments, posing potential risks to organisms. The common aquatic model organism, zebrafish (Danio rerio), are widely used to evaluate the toxicity of pesticides. In this review, we searched the Web of Science database for articles published between 2012 and 2022, using the keywords "pesticide", "zebrafish", and "toxicity", retrieving 618 publications. Furthermore, we described the main pathways by which pesticides enter aquatic environments and the fate of their residues in these environments. We systematically reviewed the toxicity effects of pesticides on zebrafish, including developmental toxicity, endocrine-disrupting effects, reproductive toxicity, neurotoxicity, immunotoxicity, and genotoxicity. Importantly, we summarized the latest research progress on the toxicity mechanism of pesticides to zebrafish based on omics technologies, including transcriptomics, metabolomics, and microbiomics. Finally, we discussed future research prospects, focusing on the combined exposure of multiple pollutants including pesticides, the risk of multigenerational exposure to pesticides, and the chronic toxicity of aquatic nanopesticides. This review provides essential data support for ecological risk assessments of pesticides in aquatic environments, and has implications for water management in the context of pesticide pollution.
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Plaguicidas , Contaminantes Químicos del Agua , Animales , Plaguicidas/toxicidad , Plaguicidas/metabolismo , Pez Cebra/metabolismo , Contaminantes Químicos del Agua/toxicidad , Contaminantes Químicos del Agua/metabolismoRESUMEN
Biothiols (GSH, Hcy, Cys) are important active sulfur substances in biological systems and widely participate in various physiological processes. The three kinds of biothiols have similar chemical structures, including the sulfhydryl group (-SH) and an amino group (-NH2), so distinguishing two or more of them simultaneously is an important challenge. Herein, a nopinone-based fluorescent probe 3-(3-((4-nitrobenzoxadiazole vinyl) nopinyl difluoride (NF-NBD) was designed to distinguish GSH and Hcy/Cys by generating different fluorescence channels with a single excitation wavelength. The nitrobenzodioxazole (NBD) was introduced in the fluorescent probe by ether bounds that can quench fluorescence and selectively discriminate GSH and Hcy/Cys. After reacting with GSH and Hcy/Cys, NF-NBD exhibited strong fluorescence (green for GSH and yellow for Hcy/Cys). NF-NBD displayed a wide linear range, low detection limit, a rapid response time, and superior selectivity for biothiols. Furthermore, NF-NBD was applied to image and distinguish different biothiols in living cells and zebrafish via different fluorescence signals at a single excitation wavelength.
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Colorantes Fluorescentes , Pez Cebra , Animales , Éteres , Éteres de Etila , FluorescenciaRESUMEN
As a virulent heavy metal ion, Hg2+ will lead to a serious threat to ecosystem and human health. In this work, we reported a chitosan-naphthalimide fluorescent probe CS-NA-ITC for specific recognition and efficient adsorption of Hg2+. CS-NA-ITC showed no fluorescence in solution state, while the fluorescence intensity increased obviously at the presence of Hg2+, accompanied by the fluorescence color becomes from colorless to bright yellow. It displayed favorable properties like low detection limit (73 nM), extensive pH detection range (5-10) and excellent anti-interference ability. The binding pattern of CS-NA-ITC to Hg2+ was verified by Job's plot, XPS analysis and FT-IR test. In addition, CS-NA-ITC was utilized to recognition of Hg2+ in actual water and soil samples and seafood products. Furthermore, the CS-NA-ITC hydrogel could be employed as an efficient Hg2+ adsorbent with good reusability, which adsorption ability was enhanced compared to chitosan hydrogel.
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Quitosano , Mercurio , Humanos , Colorantes Fluorescentes/química , Quitosano/química , Naftalimidas , Espectroscopía Infrarroja por Transformada de Fourier , Adsorción , Agua/análisis , Suelo , Ecosistema , Mercurio/química , Hidrogeles , Alimentos Marinos/análisisRESUMEN
Structures and dynamics of transmembrane (TM) receptor regions are key to understanding their signaling mechanism across membranes. Here we examine configurations of TM region dimers, assembled using the recent Martini 3 force field for coarse-grain (CG) molecular dynamics simulations. At first glance, our results show only a reasonable agreement with ab initio predictions using PREDDIMER and AlphaFold2 Multimer and with nuclear magnetic resonance (NMR)-derived structures. 5 of 11 CG TM structures are similar to the NMR structures (within <3.5 Å root-mean-square deviation [RMSD]) compared with 10 and 9 using PREDDIMER and AlphaFold2, respectively (with 8 structures of the later within 1.5 Å). Surprisingly, AlphaFold2 predictions are closer to NMR structures when the 2001 instead of 2020 database is used for training. The CG simulations reveal that alternative configurations of TM dimers readily interconvert with a predominant population. The implications for transmembrane signaling are discussed, including for the development of peptide-based pharmaceuticals.
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Simulación de Dinámica Molecular , PéptidosRESUMEN
As the most commonly used organochlorine pesticide nowadays, chlorothalonil (CHI), is ubiquitous in a natural environment and poses many adverse effects to organisms. Unfortunately, the toxicity mechanisms of CHI have not been clarified yet. This study found that the CHI based on ADI level could induce obesity in mice. In addition, CHI could induce an imbalance in the gut microbiota of mice. Furthermore, the results of the antibiotic treatment and gut microbiota transplantation experiments showed that the CHI could induce obesity in mice in a gut microbiota-dependent manner. Based on the results of targeted metabolomics and gene expression analysis, CHI could disturb the bile acids (BAs) metabolism of mice, causing the inhibition of the signal response of BAs receptor FXR and leading to glycolipid metabolism disorders in liver and epiWAT of mice. The administration of FXR agonist GW4064 and CDCA could significantly improve the CHI-induced obesity in mice. In conclusion, CHI was found to induce obesity in mice by regulating the gut microbiota and BAs metabolism via the FXR signaling pathway. This study provides evidence linking the gut microbiota and pesticides exposure with the progression of obesity, demonstrating the key role of gut microbiota in the toxic effects of pesticides.
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Microbioma Gastrointestinal , Ratones , Animales , Ácidos y Sales Biliares/metabolismo , Ácidos y Sales Biliares/farmacología , Obesidad/inducido químicamente , Obesidad/metabolismo , Hígado/metabolismo , Ratones Endogámicos C57BLRESUMEN
As one of the most abundant metal ions, Cu2+ has turned into a great threat to human health and the natural environment due to its widely utilized in various industries. In this paper, a chitosan-based fluorescent probe CTS-NA-HY for detection and adsorption of Cu2+ was rationally prepared. CTS-NA-HY exhibited a specific "turn off" fluorescence response to Cu2+ and the fluorescence color changed from bright yellow to colorless. It possessed satisfactory detection performance to Cu2+ including good selectivity and anti-interference, low detection limit (29 nM) and wide pH range (4-9). The detection mechanism was confirmed by Job's plot, X-ray photoelectron spectroscopy, FT-IR and 1H NMR analysis. Additionally, the probe CTS-NA-HY was capacity of determining Cu2+ in environmental water and soil samples. Besides, CTS-NA-HY-based hydrogel could also remove Cu2+ in aqueous solution effectively, which the ability of adsorption was greatly improved compared with original chitosan hydrogel.
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Quitosano , Naftalimidas , Humanos , Naftalimidas/química , Cobre/química , Espectroscopía Infrarroja por Transformada de Fourier , Colorantes Fluorescentes/química , Quitosano/química , Adsorción , Agua/química , Espectrometría de Fluorescencia/métodosRESUMEN
In this work, a novel coumarin derivative-modified cellulose acetate (DCB-CA) was synthesized as a fluorescent probe for highly selective and sensitive determination of CN- in food samples. The DCB-CA was synthesized by using CA as a skeleton, and the coumarin derivative as the fluorophore. The DCB-CA obtained was characterized by different methods including FTIR, SEM, 1H-NMR, TGA and UV-vis spectroscopy. The DCB-CA exhibited a significant "turn-off" fluorescence response to CN-, accompanied by a distinct fluorescence color change from bright yellow to colorless. The detection limit of CN- using DCB-CA was calculated to be 5.8 × 10-7 M, which was much lower than the threshold limit of CN- recommended by the World Health Organization (1.9 × 10-6 M). Because of the favorable solubility and processability of the CA, the DCB-CA was easily processed into different fluorescent materials including fluorescent films and coatings. The fluorescent film obtained was also applied to the selective detection of CN-. Furthermore, the DCB-CA was successfully applied to determine CN- in food samples.
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Cumarinas , Colorantes Fluorescentes , Colorantes Fluorescentes/química , Cumarinas/química , Espectrometría de Fluorescencia/métodos , Espectroscopía de Resonancia MagnéticaRESUMEN
Obesity caused by endocrine disruptors (EDCs) has become a hot topic threatening human health. Recently, Nanoselenium Siraitia grosvenorii (NSG) has been shown to have potential health-modulating uses. Based on the results of 16S rRNA sequencing and metabolomics analysis, NSG has the unique function of improving gut microbiota and inhibiting obesity. Specifically, NSG can enhance gut microbiota diversity and change their composition. A significant positive correlation exists between the liver change in lysine and the high-importance dominant species ([Ruminococcus]_gnavus, Alistipes_finegoldii, etc.). NSG metabolites analysis showed that the lysine level increased by 44.45% and showed a significantly negatively correlated with (TG, TC, Leptin, etc.). Significantly, NSG reduces the degradation of lysine metabolism in the liver and inhibits fatty acid ß-oxidation. In addition, NSG decreased Acetyl-CoA levels by 24% and regulated the downregulation of TCA genes (CS, Ogdh, Fh1, and Mdh2) and the upregulation of ketone body production genes (BDH1). NSG may have a positive effect on obesity by reducing the participation of Acetyl-CoA in the TCA cycle pathway and enhancing the ketogenic conversion of Acetyl-CoA. In conclusion, the results of this study may provide a new dietary intervention strategy for preventing endocrine disruptor-induced obesity.
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Disruptores Endocrinos , Microbioma Gastrointestinal , Humanos , Disruptores Endocrinos/toxicidad , Lisina , ARN Ribosómico 16S/genética , Acetilcoenzima A/metabolismo , Acetilcoenzima A/farmacología , Obesidad/inducido químicamente , Obesidad/tratamiento farmacológico , Redes y Vías Metabólicas , Dieta Alta en GrasaRESUMEN
Hydrogen peroxide (H2O2) is involved in many important tasks in normal cell metabolism and signaling. However, abnormal levels of H2O2 are associated with the occurrence of several diseases. Therefore, it is important to develop a new method for the detection of H2O2in vivo and in vitro. A turn-off sensor, 2,2-difluoro-4,6-bis(3-methoxy-4-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)oxy)styryl)-2H-1,3,2-dioxaborine (DFCB), based on curcumin was developed for the detection of H2O2. The DFCB, an orange-emitting sensor, was constructed by employing 2,2-difluoro-4,6-bis(4-hydroxy-3-methoxystyryl)-2H-1,3,2-dioxaborine (DFC) as the main carrier, and 2-(4-bromomethylphenyl)-4,4,5,5-tetramethyl-1,3,2-doxaborolane as the recognition site. The recognition group on the DFCB sensor could be completely cleaved by H2O2 to generate the intermediate DFC, which would lead to a colorimetric change from bright orange to light blue accompanying by a significantly quenched fluorescence, which could be seen by the naked eye. This sensor exhibited a highly specific fluorescence response to H2O2, in preference to other relevant species, with an excellent anti-interference performance. The sensor DFCB also possessed some advantages including a wide pH response range (6-11), a broad linear range (0-300 µM), and a low detection limit (1.31 µM). The sensing mechanism of the DFCB sensor for H2O2 was verified by HRMS analysis, 1H-NMR titration and DFT calculations. In addition, the use of the DFCB sensor was compatible with the fluorescence imaging of H2O2 in living cells and zebrafish.
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Curcumina , Animales , Colorimetría/métodos , Peróxido de Hidrógeno/análisis , Pez Cebra , Colorantes Fluorescentes/químicaRESUMEN
Nucleosomes drastically limit transcription factor (TF) occupancy, while pioneer transcription factors (PFs) somehow circumvent this nucleosome barrier. In this study, we compare nucleosome binding of two conserved S. cerevisiae basic helix-loop-helix (bHLH) TFs, Cbf1 and Pho4. A cryo-EM structure of Cbf1 in complex with the nucleosome reveals that the Cbf1 HLH region can electrostatically interact with exposed histone residues within a partially unwrapped nucleosome. Single-molecule fluorescence studies show that the Cbf1 HLH region facilitates efficient nucleosome invasion by slowing its dissociation rate relative to DNA through interactions with histones, whereas the Pho4 HLH region does not. In vivo studies show that this enhanced binding provided by the Cbf1 HLH region enables nucleosome invasion and ensuing repositioning. These structural, single-molecule, and in vivo studies reveal the mechanistic basis of dissociation rate compensation by PFs and how this translates to facilitating chromatin opening inside cells.