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1.
Spectrochim Acta A Mol Biomol Spectrosc ; 324: 124950, 2025 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-39133976

RESUMO

Low fluorescence under visible light excitation and catalytic activity limit many applications of graphene quantum dots in optical detection, biosensing, catalysis and biomedical. The paper reports design and synthesis of histidine, serine and folic acid-functionalized and boron and iron-doped graphene quantum dot (Fe/B-GQD-HSF). The Fe/B-GQD-HSF shows excellent fluorescence behavior and peroxidase-like activity. Excitation of 330 nm ultraviolet light produces the strongest blue fluorescence and excitation of 480 nm visible light produces the strongest yellow fluorescence. The specific activity reaches 92.67 U g-1, which is higher than that of other graphene quantum dots. The Fe/B-GQD-HSF can catalyze oxidation of 3,3',5,5'-tetramethylbenzidine with H2O2 to form blue compound. Based on this, it was used for colorimetric and fluorescence detection of H2O2. The absorbance at 652 nm linearly increases with the increase of H2O2 concentration between 0.5 and 100 µM with detection limit of 0.43 µM. The fluorescence signal linearly decreases with the increase of H2O2 concentration between 0.05 and 100 µM with detection limit of 0.035 µM. The analytical method has been satisfactorily applied in detection of H2O2 in food. The study also paves one way for design and synthesis of functional graphene quantum dots with ideal fluorescence behavior and catalytic activity.


Assuntos
Boro , Colorimetria , Ácido Fólico , Grafite , Histidina , Peróxido de Hidrogênio , Ferro , Pontos Quânticos , Serina , Pontos Quânticos/química , Grafite/química , Peróxido de Hidrogênio/análise , Peróxido de Hidrogênio/química , Colorimetria/métodos , Ácido Fólico/análise , Ácido Fólico/química , Ferro/análise , Ferro/química , Boro/química , Histidina/análise , Histidina/química , Serina/análise , Serina/química , Espectrometria de Fluorescência/métodos , Limite de Detecção , Análise de Alimentos/métodos , Peroxidase/química , Peroxidase/metabolismo , Catálise
2.
J Environ Sci (China) ; 148: 198-209, 2025 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-39095157

RESUMO

Norfloxacin is widely used owing to its strong bactericidal effect on Gram-negative bacteria. However, the residual norfloxacin in the environment can be biomagnified via food chain and may damage the human liver and delay the bone development of minors. Present work described a reliable and sensitive smartphone colorimetric sensing system based on cobalt-doped Fe3O4 magnetic nanoparticles (Co-Fe3O4 MNPs) for the visual detection of norfloxacin. Compared with Fe3O4, Co-Fe3O4 MNPs earned more remarkably peroxidase-like activity and TMB (colorless) was rapidly oxidized to oxTMB (blue) with the presence of H2O2. Interestingly, the addition of low concentration of norfloxacin can accelerate the color reaction process of TMB, and blue deepening of the solution can be observed with the naked eye. However, after adding high concentration of norfloxacin, the activity of nanozyme was inhibited, resulting in the gradual fading of the solution. Based on this principle, a colorimetric sensor integrated with smartphone RGB mode was established. The visual sensor exhibited good linearity for norfloxacin monitoring in the range of 0.13-2.51 µmol/L and 17.5-100 µmol/L. The limit of visual detection was 0.08 µmol/L. In the actual water sample analysis, the spiked recoveries of norfloxacin were over the range of 95.7%-104.7 %. These results demonstrated that the visual sensor was a convenient and fast method for the efficient and accurate detection of norfloxacin in water, which may have broad application prospect.


Assuntos
Cobalto , Colorimetria , Norfloxacino , Smartphone , Poluentes Químicos da Água , Norfloxacino/análise , Colorimetria/métodos , Cobalto/análise , Cobalto/química , Poluentes Químicos da Água/análise , Antibacterianos/análise , Peroxidase , Limite de Detecção
3.
Mediators Inflamm ; 2024: 8847492, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39238946

RESUMO

The pathogenesis of acute lung injury is complex. Studies have demonstrated the role of neutrophil extracellular traps (NETs) in the process of lipopolysaccharide (LPS)-induced acute lung injury (ALI). However, the underlying mechanism remains unclear. In this study, the regulation of Nrf2 in the formation of NETs, which was pathogenic in LPS-induced ALI, was identified by analyzing the levels of Cit-H3, lung function, lung tissue pathology, lung wet/dry ratio, the inflammatory cells, cytokines and proteins in the bronchoalveolar lavage fluid (BALF) and in addition, the activity of lung myeloperoxidase (MPO) was also measured. Results showed that the levels of Cit-H3 measured by western blot in Nrf2-knockout (KO) mice were higher compared with the WT mice after LPS stimulation. To further investigate the NETs formation was pathogenic during LPS-induced ALI, the Nrf2-KO mice were treated with DNase I. Results showed that DNase I improved lung function and lung tissue pathology and significantly reduced lung wet/dry ratio and proteins in the BALF. Besides, DNase I also attenuated the infiltration of inflammatory cells and the cytokines (TNF-α, IL-1ß) production in the BALF and the activity of lung MPO. Therefore, these results together indicate that Nrf2 may intervene in the release of NETs during LPS-induced ALI in mice.


Assuntos
Lesão Pulmonar Aguda , Líquido da Lavagem Broncoalveolar , Armadilhas Extracelulares , Lipopolissacarídeos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 2 Relacionado a NF-E2 , Animais , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/induzido quimicamente , Fator 2 Relacionado a NF-E2/metabolismo , Camundongos , Armadilhas Extracelulares/metabolismo , Líquido da Lavagem Broncoalveolar/química , Masculino , Peroxidase/metabolismo , Neutrófilos/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Interleucina-1beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Desoxirribonuclease I/metabolismo , Citocinas/metabolismo , Western Blotting
4.
Mikrochim Acta ; 191(10): 599, 2024 09 14.
Artigo em Inglês | MEDLINE | ID: mdl-39276245

RESUMO

A colorimetric sensor for the rapid and sensitive detection of GSH was developed. The hydrothermal method was utilized to synthesize chitosan-stabilized gold nanoparticles (CS-AuNPs). The synthesized CS-AuNPs were characterized by UV-vis absorption spectroscopy, transmission electron microscopy (TEM), X-ray diffractograms (XRD), and Fourier transform infrared spectroscopy (FTIR). The CS-AuNPs are well-dispersed and possess a spherical shape with an average particle size of 10.05 ± 2.26 nm in aqueous solution. They show an intrinsic peroxidase-like activity, which could efficiently catalyze the decomposition of H2O2 to produce •OH radicals. These radicals then oxidized 3, 3´, 5, 5´-tetramethylbenzidine (TMB), resulting in the formation of the blue oxidized product oxTMB, observed a visible color change (from colorless to blue), and oxTMB had an obvious absorption peak at 652 nm. The presence of GSH could inhibit the peroxidase-like activity of CS-AuNPs, thereby reducing the formation of oxTMB. The solution's blue hue underwent a reduction in absorption intensity. Based on this fact, a novel and sensitive colorimetric sensor for detection of GSH was constructed. Under optimal conditions, the results of detection had an excellent linear relationship between the concentration of GSH and ∆A within the range 0.5 ~ 50.0 × 10-6 mol/L. The limit of detection (LOD) for GSH was 2.10 × 10-7 mol/L, which was much lower than those in most previous works. Furthermore, for detection in real human serum samples, the recoveries of GSH and the relative standard deviations (RSD) in the serum were in the range 98.40 ~ 103.32% and 1.85 ~ 3.54%, respectively. Thus, this visual colorimetric method has good precision and can be used for GSH detection in practical applications, promising in the fields of bioanalysis and illness diagnostics.


Assuntos
Quitosana , Colorimetria , Glutationa , Ouro , Limite de Detecção , Nanopartículas Metálicas , Ouro/química , Humanos , Colorimetria/métodos , Quitosana/química , Nanopartículas Metálicas/química , Glutationa/sangue , Glutationa/química , Peróxido de Hidrogênio/química , Benzidinas/química , Peroxidase/química
5.
Acta Cir Bras ; 39: e395524, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39230095

RESUMO

PURPOSE: To investigate the impact of the Chinese medicine compound Ento-PB on oxazolone (OXZ)-induced ulcerative colitis (UC) in rats. METHODS: UC rats induced by OXZ were treated with Ento-PB. The damage to the colon was assessed using several measures, including the disease activity index (DAI), colon length, colon weight/length ratio, colonic mucosal damage index, and histological score. The levels of interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-13 (IL-13), epidermal growth factor (EGF), inducible nitric oxide synthase, and total nitric oxide synthase (tNOS) in rat serum, as well as the levels of tumor necrosis factor-α (TNF-α) and myeloperoxidase (MPO) in rat colon tissue, were determined using enzyme-linked immunosorbent assay and conventional kits. RESULTS: After being treated with Ento-PB, the DAI score and macroscopic lesion score of OXZ-induced UC rats were significantly reduced. Ento-PB prevented the shortening of rat colons, reduced the ratio of colon weight to length, and improved colon tissue lesions. Meanwhile, Ento-PB could significantly inhibit the activities of proinflammatory cytokines TNF-α, IL-13, and MPO, as well as tNOS and iNOS, while upregulating the expression of anti-inflammatory cytokines IL-4 and IL-10. Moreover, a significant increase in the expression level of EGF was observed in UC rats treated with Ento-PB, indicating that Ento-PB could enhance the repair of damaged intestinal epithelial tissue. CONCLUSIONS: Ento-PB demonstrates significant anti-UC activities in OXZ-induced UC rats by regulating the expression levels of inflammatory factors and promoting the repair of colon tissue. This study provides scientific evidence to support the further development of Ento-PB.


Assuntos
Colite Ulcerativa , Colo , Oxazolona , Peroxidase , Animais , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/patologia , Masculino , Colo/efeitos dos fármacos , Colo/patologia , Colo/metabolismo , Peroxidase/análise , Peroxidase/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Modelos Animais de Doenças , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/metabolismo , Ratos Sprague-Dawley , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/patologia , Mucosa Intestinal/metabolismo , Ratos , Ensaio de Imunoadsorção Enzimática , Fator de Crescimento Epidérmico/análise , Citocinas/metabolismo , Interleucina-13/análise , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico Sintase Tipo II/análise , Reprodutibilidade dos Testes , Resultado do Tratamento
6.
Talanta ; 280: 126785, 2024 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-39217709

RESUMO

In the present research, Fe-based metal-organic frameworks (MIL-101(Fe)-NH2) nanoparticles were synthesized by simple solvothermal methods and used to assay Cr(Ⅵ). The MIL-101(Fe)-NH2 performs dual functions: the 2-aminoterephthalic acid (NH2-BDC) ligand endows a strong fluorescence emission, and the Fe metal nodes are able to facilitate the oxidation of 3,3',5,5'- tetramethylbenzidine (TMB) directly, resulting in the generation of oxidized-TMB (ox-TMB). Our research results showed that reducing agents such as ascorbic acid (AA) can collapse the structures of MIL-101(Fe)-NH2 because of the reduction of Fe3+ by AA, resulting in release of NH2-BDC. In the presence of Cr(Ⅵ), the fluorescence intensity of the MIL-101(Fe)-NH2 + AA system will be decreased due to the competitive reduction of Fe3+ and Cr(Ⅵ). Nevertheless, Cr(Ⅵ) can significantly accelerate the oxidation of TMB by MIL-101(Fe)-NH2 as it boosts the electron transfer rate between Fe3+ and Fe2+. Therefore, a fluorescent/colorimetric dual-mode platform was developed for the detection of Cr(Ⅵ) with an extensive linear range (7.5-750 µg/L and 13.3-1000 µg/L) as well as a remarkably low detection limit (0.99 µg/L and 1.98 µg/L). This MOF with the ability to release ligands not only provides inspiration for the design of new luminescent materials, but also offers a novel and reliable solution for the detection of Cr(Ⅵ).


Assuntos
Cromo , Colorimetria , Corantes Fluorescentes , Estruturas Metalorgânicas , Cromo/análise , Cromo/química , Estruturas Metalorgânicas/química , Corantes Fluorescentes/química , Colorimetria/métodos , Limite de Detecção , Benzidinas/química , Oxirredução , Ferro/química , Espectrometria de Fluorescência/métodos , Peroxidase/química , Peroxidase/metabolismo , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/química
7.
Sci Rep ; 14(1): 21231, 2024 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-39261545

RESUMO

The study assessed selected parameters of redox status in the plasma of patients suffering from high myopia (HM). Thirty-five children with mean age 13.7 ± 2.7 years with HM and 40 healthy children were included. Plasma redox status parameters were determined using colorimetric kits. The levels of retinol, α-tocopherol and coenzyme Q10 were determined with a high-performance liquid chromatograph. Negative correlations were observed between the concentrations of retinol and the axial length of the eye (r = - 0.514 p < 0.001). Increased myeloperoxidase (MPO) activity (p < 0.018), and decreased concentrations of retinol (p < 0.001) and α-tocopherol (p < 0.023) in patients with HM and the axial length of the eye > 26 mm compared to controls were established. Significantly lower retinol and α-tocopherol concentrations were found in patients with the axial length of the eye > 26 mm compared to those with the axial length of the eye ≤ 26 mm (p < 0.001, p < 0.021, respectively). Increased MPO activity in advanced stages of HM may confirm an inflammatory process in HM patients. Reduced retinol and α-tocopherol concentrations and their link to disease progression indicate a need for monitoring their levels and supplementation in children with HM.


Assuntos
Miopia , Peroxidase , Vitamina A , alfa-Tocoferol , Humanos , alfa-Tocoferol/sangue , Peroxidase/sangue , Vitamina A/sangue , Masculino , Feminino , Criança , Adolescente , Miopia/sangue , Miopia/metabolismo , Estudos de Casos e Controles
8.
Int J Mol Sci ; 25(17)2024 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-39273368

RESUMO

This study aimed to compare several potential mouthrinse biomarkers for periodontitis including active matrix-metalloproteinase-8 (aMMP-8), total MMP-8, and other inflammatory biomarkers in diagnosing and monitoring the effects of nonsurgical periodontal therapy. Thirteen patients with stage III/IV periodontitis were recruited, along with thirteen periodontally and systemically healthy controls. These 13 patients were representative of the number of outpatients visiting any dentist in a single day. Full-mouth clinical periodontal parameters and biomarkers (the aMMP-8 point-of-care-test [POCT], total MMP-8, tissue inhibitor of MMPs (TIMP)-1, the aMMP-8 RFU activity assay, Myeloperoxidase, PMN elastase, calprotectin, and interleukin-6) were recorded at baseline and after nonsurgical therapy at 6 weeks. The aMMP-8 POCT was the most efficient and precise discriminator, with a cut-off of 20 ng/mL found to be optimal. Myeloperoxidase, MMP-8's oxidative activator, was also efficient. Following closely in precision was the aMMP-8 RFU activity assay and PMN elastase. In contrast, the total MMP-8 assay and the other biomarkers were less efficient and precise in distinguishing patients with periodontitis from healthy controls. aMMP-8, MPO, and PMN elastase may form a proteolytic and pro-oxidative tissue destruction cascade in periodontitis, potentially representing a therapeutic target. The aMMP-8 chair-side test with a cut-off of 20 ng/mL was the most efficient and precise discriminator between periodontal health and disease. The aMMP-8 POC test can be effectively used by dental professionals in their dental practices in online and real-time diagnoses as well as in monitoring periodontal disease and educating and encouraging good oral practices among patients.


Assuntos
Biomarcadores , Metaloproteinase 8 da Matriz , Periodontite , Humanos , Metaloproteinase 8 da Matriz/metabolismo , Periodontite/diagnóstico , Periodontite/terapia , Periodontite/metabolismo , Feminino , Masculino , Pessoa de Meia-Idade , Adulto , Peroxidase/metabolismo , Sistemas Automatizados de Assistência Junto ao Leito , Elastase de Leucócito/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo
9.
Anal Chim Acta ; 1326: 343150, 2024 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-39260918

RESUMO

BACKGROUND: To minimize the impact of pesticide residues in food on human health, it is necessary to enhance their detection. Recently, many nanozyme-based colorimetric methods for pesticides detection have been developed, however, they often required the assistance of natural enzymes, which made the process and result of methods susceptible to the stability and activity of natural enzymes. To overcome these drawbacks, methods for direct detection of pesticides using nanozymes have been developed, and there are few studies in this field currently. Thus, it is of great research and practical significance to develop more nanozymes-based colorimetric methods for direct detection of pesticides. RESULTS: Dual colorimetric platforms based on Os-Rh nanozyme with excellent peroxidase-like activity were constructed for directly detection of glyphosate in this work. Results showed that glyphosate was able to sensitively and selectively inhibit the peroxidase-like activity of Os-Rh nanozyme through hindering the decomposition of H2O2 by Os-Rh nanozyme to produce HO∙. Based on this, the dual colorimetric platforms achieved highly sensitive detection for glyphosate over a wide linear concentration range (50-1000 µg L-1 in solution platform and 200-1000 µg L-1 in paper platform), with the detection limits of 28.37 µg L-1 in solution platform and 400 µg L-1 (naked-eye detection limit)/123.25 µg L-1 (gray scale detection limit) in paper platform, respectively. Moreover, the dual colorimetric platforms possessed satisfactory reliability and accuracy for practical applications, and has been successfully applied to the detection of real samples with the spiked recoveries of 92.78-102.75 % and RSD of 1.17-3.88 %. SIGNIFICANCE: The dual colorimetric platforms for glyphosate direct detection based on Os-Rh nanozyme developed in this work not only owned considerable practical application potential, but also could provide more inspirations and ideas for the rational design and development of colorimetric sensing methods for the rapid detection of pesticides based on nanozymes.


Assuntos
Colorimetria , Glicina , Glifosato , Colorimetria/métodos , Glicina/análogos & derivados , Glicina/análise , Glicina/química , Peroxidase/metabolismo , Peroxidase/química , Limite de Detecção , Peróxido de Hidrogênio/química
10.
Medicine (Baltimore) ; 103(36): e39494, 2024 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-39252215

RESUMO

Sepsis is an infection-induced systemic inflammatory response syndrome. Immune regulation plays a crucial role in sepsis. We looked into the link between immune effector-related proteins and sepsis in this study by using both univariate and multivariate Mendelian randomization (MR) analyses. We accessed and collected data from the Integrative Epidemiology Unit's Open About Sepsis genome-wide association study database. The 6 immune effector-associated proteins each contained 10,534,735 single-nucleotide polymorphisms from 3301 samples. Using the weighted median, MR-Egger, simplex, inverse-variance weighting, and weighted mode methods, univariate MR then investigated the link between complement factor H-related protein-5 (CFHR5), Fc epsilon receptor II (FCER2), granzyme B (GZMB), major histocompatibility complex, class II, DQ alpha (HLA-DQA2), mannose-binding lectin 2 (MBL2), or myeloperoxidase (MPO) and sepsis. In the inverse-variance weighted results, the P values of all 6 immune effector-related proteins were <0.05, suggesting a possible causal relationship between them and sepsis. MBL2 (odds ratio [OR] = 1.046) was a risk factor for sepsis, while the other proteins (FCER2: OR = 0.922; GZMB: OR = 0.908; CFHR5: OR = 0.858; HLA-DQA2: OR = 0.896; MPO: OR = 0.875) were safety factors. By revealing a causal link between sepsis and CFHR5, FCER2, GZMB, HLA-DQA2, MBL2, or MPO, our study offers an essential resource for additional investigations on the subject.


Assuntos
Estudo de Associação Genômica Ampla , Lectina de Ligação a Manose , Análise da Randomização Mendeliana , Polimorfismo de Nucleotídeo Único , Sepse , Humanos , Sepse/genética , Sepse/imunologia , Lectina de Ligação a Manose/genética , Granzimas/genética , Peroxidase/genética , Peroxidase/imunologia , Fatores de Risco , Predisposição Genética para Doença , Receptores Fc/genética
11.
Plant Signal Behav ; 19(1): 2400451, 2024 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-39235999

RESUMO

Lead is one of the major environmental pollutants which is highly toxic to plants and living beings. The current investigation thoroughly evaluated the synergistic effects of oxalic acid (OA) and salicylic acid (SA) on Zea mays L. plants subjected to varying durations (15, 30, 30, and 45 days) of lead (Pb) stress. Besides, the effects of oxalic acid (OA) combined with salicylic acid (SA) for different amino acids at various periods of Pb stress were also investigated on Zea mays L. The soil was treated with lead nitrate Pb (NO3)2 (0.5 mM) to induce Pb stress while the stressed plants were further treated using oxalic acid (25 mg/L), salicylic acid (25 mg/L), and their combination OA + SA (25 mg/L each). Measurements of protein content, malondialdehyde (MDA) levels, guaiacol peroxidase (GPOX) activity, catalase (CAT) activity, GSH content, and Pb concentration in maize leaves were done during this study. MDA levels increased by 71% under Pb stress, while protein content decreased by 56%, GSH content by 35%, and CAT activity by 46%. After treatment with SA, OA, and OA+SA, there was a significant reversal of these damages, with the OA+SA combination showing the highest improvement. Specifically, OA+SA treatment led to a 45% increase in protein content and a 39% reduction in MDA levels compared to Pb treatment alone. Moreover, amino acid concentrations increased by 68% under the Pb+OA+SA treatment, reflecting the most significant recovery (p < 0.0001).


Assuntos
Aminoácidos , Chumbo , Malondialdeído , Ácido Oxálico , Ácido Salicílico , Estresse Fisiológico , Zea mays , Zea mays/efeitos dos fármacos , Zea mays/metabolismo , Chumbo/toxicidade , Ácido Oxálico/metabolismo , Ácido Oxálico/farmacologia , Ácido Salicílico/farmacologia , Aminoácidos/metabolismo , Malondialdeído/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Catalase/metabolismo , Peroxidase/metabolismo , Glutationa/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Sinergismo Farmacológico , Proteínas de Plantas/metabolismo
12.
Mikrochim Acta ; 191(10): 588, 2024 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-39256210

RESUMO

Different morphological Cu2O nanoparticles including cube, truncated cube, and octahedron were successfully prepared by a selective surface stabilization strategy. The prepared cube Cu2O exhibited superior peroxidase-like activity over the other two morphological Cu2O nanoparticles, which can readily oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to form visually recognizable color signals. Consequently, a sensitive and simple colorimetric biosensor was proposed for deoxynivalenol (DON) detection. In this biosensor, the uniform cube Cu2O was employed as the vehicle to label the antibody for the recognition of immunoreaction. The sensing strategy showed a detection limit as low as 0.01 ng/mL, and a wide linear range from 2 to 100 ng/mL. Concurrently, the approximate DON concentration can be immediately and conveniently observed by the vivid color changes. Benefiting from the high sensitivity and selectivity of the designed biosensor, the detection of DON in wheat, corn, and tap water samples was achieved, suggesting the bright prospect of the biosensor for the convenient and intuitive detection of DON in actual samples.


Assuntos
Benzidinas , Técnicas Biossensoriais , Colorimetria , Cobre , Limite de Detecção , Nanopartículas Metálicas , Tricotecenos , Zea mays , Tricotecenos/análise , Tricotecenos/imunologia , Colorimetria/métodos , Cobre/química , Técnicas Biossensoriais/métodos , Benzidinas/química , Zea mays/química , Nanopartículas Metálicas/química , Triticum/química , Peroxidase/química , Anticorpos Imobilizados/imunologia , Contaminação de Alimentos/análise
13.
Anal Chem ; 96(36): 14590-14597, 2024 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-39183481

RESUMO

Hemin/G-quadruplex (hG4) complexes are frequently used as artificial peroxidase-like enzymatic systems (termed G4 DNAzymes) in many biosensing applications, in spite of a rather low efficiency, notably in terms of detection limits. To tackle this issue, we report herein a strategy in which hemin is chemically modified with the amino acids found in the active site of parent horseradish peroxidase (HRP), with the aim of recreating an environment conducive to high catalytic activity. When hemin is conjugated with a single arginine, it associates with G4 to create an arginine-hemin/G4 (R-hG4) DNAzyme that exhibits improved catalytic performances, characterized by kinetic analysis and DFT calculations. The practical relevance of this system was demonstrated with the implementation of biosensing assays enabling the chemiluminescent detection of G4-containing DNA and colorimetry detection of the flap endonuclease 1 (FEN1) enzyme with a high efficiency and sensitivity. Our results thus provide a guide for future enzyme engineering campaigns to create ever more efficient peroxidase-mimicking DNA-based systems.


Assuntos
Arginina , DNA Catalítico , Quadruplex G , Hemina , Hemina/química , DNA Catalítico/química , DNA Catalítico/metabolismo , Arginina/química , Arginina/metabolismo , Técnicas Biossensoriais/métodos , Peroxidase/química , Peroxidase/metabolismo , Peroxidase do Rábano Silvestre/química , Peroxidase do Rábano Silvestre/metabolismo , Limite de Detecção , Colorimetria , Teoria da Densidade Funcional
14.
Innate Immun ; 30(5): 82-89, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-39090856

RESUMO

Cardiovascular diseases (CVDs) linked to atherosclerosis remains the leading cause of death worldwide. Atherosclerosis is primarily caused by the accumulation of oxidized forms of low density lipoprotein (LDL) in macrophages (MΦs) in the subendothelial layer of arteries leading to foam cell and fatty streak formation. Many studies suggest that LDL that is modified by myeloperoxidase (MPO) is a key player in the development of atherosclerosis. MΦs can adopt a variety of functional phenotypes that include mainly the proinflammatory M1 and the anti-inflammatory M2 MΦ phenotypes which are both implicated in the process of atherogenesis. In fact, MΦs that reside in atherosclerostic lesions were shown to express a variety of phenotypes ranging between the M1- and M2 MΦ types. Recently, we pointed out the involvement of MPO oxidized-LDL (Mox-LDL) in increasing inflammation in MΦs by reducing their secretion of IL-10. Since little is known about Mox-LDL-mediated pro-atherosclerostic responses in MΦs, our study aimed at analyzing the in vitro effects of Mox-LDL at this level through making use of the well-established model of human THP-1-derived Mφs. Our results demonstrate that Mox-LDL has no effect on apoptosis, reactive oxygen species (ROS) generation and cell death in our cell model; yet, interestingly, our results show that Mox-LDL is significantly engulfed at a higher rate in the different MΦ subtypes supporting its key role in foam cell formation during the progression of the disease as well as previous data that were generated using another primary MΦ cell model of atherosclerosis.


Assuntos
Aterosclerose , Lipoproteínas LDL , Macrófagos , Peroxidase , Espécies Reativas de Oxigênio , Humanos , Lipoproteínas LDL/metabolismo , Peroxidase/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Aterosclerose/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Apoptose , Células THP-1 , Células Espumosas/metabolismo , Interleucina-10/metabolismo , Inflamação
15.
Int J Mol Sci ; 25(16)2024 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-39201490

RESUMO

Atherosclerosis is a chronic inflammatory disease that involves modified low-density lipoproteins (LDL) which play a pivotal role in the initiation and progression of the disease. Myeloperoxidase oxidized LDL (Mox-LDL) is considered to be the most patho-physiologically relevant type of modified LDL and has been reported to be ubiquitously present in atheroma plaques of patients with atherosclerosis. Besides its involvement in the latter disease state, Mox-LDL has also been shown to be implicated in the pathogenesis of various illnesses including sleep disorders, which are in turn associated with heart disease and depression in many intricate ways. Meanwhile, we have recently shown that lox-1-mediated Mox-LDL signaling modulates neuroserpin activity in endothelial cells, which could have major implications that go beyond the pathophysiology of stroke and cerebrovascular disease (CD). Of note is that tissue plasminogen activator (tPA), which is the main target of neuroserpin in the brain, has a crucial function in the processing of brain-derived neurotrophic factor (BDNF) into its mature form. This factor is known to be involved in major depressive disorder (MDD) development and pathogenesis. Since tPA is more conventionally recognized as being involved in fibrinolytic mechanisms, and its effect on the BDNF system in the context of MDD is still not extensively studied, we speculate that any Mox-LDL-driven change in the activity of tPA in patients with atherosclerosis may lead to a decrease in the production of mature BDNF, resulting in impaired neural plasticity and depression. Deciphering the mechanisms of interaction between those factors could help in better understanding the potentially overlapping pathological mechanisms that regulate disease processes in CD and MDD, supporting the possibility of novel and common therapeutic opportunities for millions of patients worldwide.


Assuntos
Aterosclerose , Lipoproteínas LDL , Peroxidase , Humanos , Aterosclerose/metabolismo , Lipoproteínas LDL/metabolismo , Peroxidase/metabolismo , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Depressão/metabolismo , Neuroserpina , Receptores Depuradores Classe E/metabolismo , Transtorno Depressivo Maior/metabolismo
16.
ACS Appl Mater Interfaces ; 16(34): 45371-45382, 2024 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-39140178

RESUMO

The self-assembly of proteins and peptides into fibrillar amyloid aggregates is a highly promising route to define the next generation of functional nanomaterials. Amyloid fibrils, traditionally associated with neurodegenerative diseases, offer exceptional conformational and chemical stability and mechanical properties, and resistance to degradation. Here, we report the development of catalytic amyloid nanomaterials through the conjugation of a miniaturized artificial peroxidase (FeMC6*a) to a self-assembling amyloidogenic peptide derived from human transthyretin, TTR(105-115), whose sequence is YTIAALLSPYS. Our synthetic approach relies on fast and selective click ligation upon proper modification of both the peptide and FeMC6*a, leading to TTRLys108@FeMC6*a. Mixing unmodified TTR(105-115) with TTRLys108@FeMC6*a allowed the generation of enzyme-loaded amyloid fibrils, namely, FeMC6*a@fibrils. Catalytic studies, performed in aqueous solution at nearly neutral pH, using ABTS as a model substrate and H2O2 as the oxidizing agent revealed that the enzyme retains its catalytic activity. Moreover, the activity was found to depend on the TTRLys108@FeMC6*a/unmodified TTR(105-115) peptide ratio. In particular, those with the 2:100 ratio showed the highest activity in terms of initial rates and substrate conversion among the screened nanoconjugates and compared to the freely diffusing enzyme. Finally, the newly developed nanomaterials were integrated into a flow system based on a polyvinylidene difluoride membrane filter. Within this flow-reactor, multiple reaction cycles were performed, showcasing the reusability and stability of the catalytic amyloids over extended periods, thus offering significantly improved characteristics compared to the isolated FeMC6*a in the application to a number of practical scenarios.


Assuntos
Amiloide , Nanoestruturas , Pré-Albumina , Amiloide/química , Nanoestruturas/química , Catálise , Humanos , Pré-Albumina/química , Pré-Albumina/metabolismo , Peróxido de Hidrogênio/química , Peroxidase/química , Peroxidase/metabolismo , Heme/química
17.
Talanta ; 279: 126639, 2024 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-39094531

RESUMO

In this paper, an ultra-small-sized CuOx/GDYO nanozyme in situ grown on ITO glass was rationally synthesized from mixed precursors of graphdiyne oxide (GDYO) and copper based infinite coordination polymer (Cu-ICP, consisting of Cu ions and two organic ligands 3,5-di-tert-butylcatechol and 1,4-bis(imidazole-1-ylmethyl)benzene) via mild and simple electrochemical strategy. On one hand, the preferential electro-reduction of Cu-ICP enabled the formation of ultra-small CuOx with Cu(I) as the main component and avoided the loss of oxygen-containing functional groups and defects on the surface of GDYO; on the other hand, GDYO can also serve as electroless reductive species to facilitate the electrochemical deposition of CuOx and turn itself to a higher oxidation state with more exposed functional groups and defects. This one-stone-two-birds electrochemical strategy empowered CuOx/GDYO nanozyme with superior peroxidase-mimicking activity and robust anchoring stability on ITO glass, thus enabled further exploration of the portable device with availability for point-of-use applications. Based on the organophosphorus pesticides (OPs) blocked acetylcholinesterase (AChE) activity, the competitive redox reaction was regulated to initiate the chromogenic reaction of 3,3',5,5'-tetramethylbenzidine (TMB) catalyzed by CuOx/GDYO peroxidase-like nanozyme, which laid out a foundation for the detection of OPs (with chlorpyrifos as an example). With a detection of limit low to 0.57 nM, the OPs residues during agricultural production can be directly monitored by the portable device we developed.


Assuntos
Colorimetria , Cobre , Técnicas Eletroquímicas , Praguicidas , Colorimetria/métodos , Cobre/química , Praguicidas/análise , Praguicidas/química , Técnicas Eletroquímicas/métodos , Grafite/química , Limite de Detecção , Peroxidase/metabolismo , Peroxidase/química , Nanoestruturas/química , Acetilcolinesterase/metabolismo , Acetilcolinesterase/química , Organofosfatos/química , Organofosfatos/análise , Benzidinas/química
18.
Int J Mol Sci ; 25(15)2024 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-39125806

RESUMO

Cytochrome c (CytC), a one-electron carrier, transfers electrons from complex bc1 to cytochrome c oxidase (CcO) in the electron-transport chain. Electrostatic interaction with the partners, complex bc1 and CcO, is ensured by a lysine cluster near the heme forming the Universal Binding Site (UBS). We constructed three mutant variants of mitochondrial CytC with one (2Mut), four (5Mut), and five (8Mut) Lys->Glu substitutions in the UBS and some compensating Glu->Lys substitutions at the periphery of the UBS for charge compensation. All mutants showed a 4-6 times increased peroxidase activity and accelerated binding of cyanide to the ferric heme of CytC. In contrast, decomposition of the cyanide complex with ferrous CytC, as monitored by magnetic circular dichroism spectroscopy, was slower in mutants compared to WT. Molecular dynamic simulations revealed the increase in the fluctuations of Cα atoms of individual residues of mutant CytC compared to WT, especially in the Ω-loop (70-85), which can cause destabilization of the Fe…S(Met80) coordination link, facilitation of the binding of exogenous ligands cyanide and peroxide, and an increase in peroxidase activity. It was found that only one substitution K72E is enough to induce all these changes, indicating the significance of K72 and the Ω-loop (70-85) for the structure and physiology of mitochondrial CytC. In this work, we also propose using a ferro-ferricyanide buffer as a substrate to monitor the peroxidase activity of CytC. This new approach allows us to determine the rate of peroxidase activity at moderate (200 µM) concentrations of H2O2 and avoid complications of radical formation during the reaction.


Assuntos
Citocromos c , Simulação de Dinâmica Molecular , Sítios de Ligação , Ligantes , Citocromos c/metabolismo , Citocromos c/química , Citocromos c/genética , Peroxidase/metabolismo , Peroxidase/química , Peroxidase/genética , Substituição de Aminoácidos , Ligação Proteica , Cianetos/metabolismo , Cianetos/química , Animais , Heme/metabolismo , Heme/química , Mutação
19.
Sci Rep ; 14(1): 17944, 2024 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-39095388

RESUMO

This study demonstrates that root-associated Kosakonia oryziphila NP19, isolated from rice roots, is a promising plant growth-promoting bioagent and biopesticide for combating rice blast caused by Pyricularia oryzae. In vitro experiments were conducted on fresh leaves of Khao Dawk Mali 105 (KDML105) jasmine rice seedlings. The results showed that NP19 effectively inhibited the germination of P. oryzae fungal conidia. Fungal infection was suppressed across three different treatment conditions: rice colonized with NP19 and inoculated by fungal conidia, a mix of NP19 and fungal conidia concurrently inoculated on the leaves, and fungal conidia inoculation first followed by NP19 inoculation after 30 h. Additionally, NP19 reduced fungal mycelial growth by 9.9-53.4%. In pot experiments, NP19 enhanced the activities of peroxidase (POD) and superoxide dismutase (SOD) by 6.1-63.0% and 3.0-67.7%, respectively, indicating a boost in the plant's defense mechanisms. Compared to the uncolonized control, the NP19-colonized rice had 0.3-24.7% more pigment contents, 4.1% more filled grains per panicle, 26.3% greater filled grain yield, 34.4% higher harvest index, and 10.1% more content of the aroma compound 2-acetyl-1-pyrroline (2AP); for rice colonized with NP19 and infected with P. oryzae, these increases were 0.2-49.2%, 4.6%, 9.1%, 54.4%, and 7.5%, respectively. In field experiments, blast-infected rice that was colonized and/or inoculated with NP19 treatments had 15.1-27.2% more filled grains per panicle, 103.6-119.8% greater filled grain yield, and 18.0-35.8% higher 2AP content. A higher SOD activity (6.9-29.5%) was also observed in the above-mentioned rice than in the blast-infected rice that was not colonized and inoculated with NP19. Following blast infection, NP19 applied to leaves decreased blast lesion progression. Therefore, K. oryziphila NP19 was demonstrated to be a potential candidate for use as a plant growth-promoting bioagent and biopesticide for suppressing rice blast.


Assuntos
Oryza , Doenças das Plantas , Oryza/microbiologia , Oryza/crescimento & desenvolvimento , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Raízes de Plantas/microbiologia , Raízes de Plantas/crescimento & desenvolvimento , Esporos Fúngicos , Folhas de Planta/microbiologia , Ascomicetos/patogenicidade , Plântula/microbiologia , Plântula/crescimento & desenvolvimento , Agentes de Controle Biológico/farmacologia , Peroxidase/metabolismo
20.
Nat Commun ; 15(1): 6888, 2024 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-39134525

RESUMO

Constructing atom-pair engineering and improving the activity of metal single-atom nanozyme (SAzyme) is significant but challenging. Herein, we design the atom-pair engineering of Zn-SA/CNCl SAzyme by simultaneously constructing Zn-N4 sites as catalytic sites and Zn-N4Cl1 sites as catalytic regulator. The Zn-N4Cl1 catalytic regulators effectively boost the peroxidase-like activities of Zn-N4 catalytic sites, resulting in a 346-fold, 1496-fold, and 133-fold increase in the maximal reaction velocity, the catalytic constant and the catalytic efficiency, compared to Zn-SA/CN SAzyme without the Zn-N4Cl1 catalytic regulator. The Zn-SA/CNCl SAzyme with excellent peroxidase-like activity effectively inhibits tumor cell growth in vitro and in vivo. The density functional theory (DFT) calculations reveal that the Zn-N4Cl1 catalytic regulators facilitate the adsorption of *H2O2 and re-exposure of Zn-N4 catalytic sites, and thus improve the reaction rate. This work provides a rational and effective strategy for improving the peroxidase-like activity of metal SAzyme by atom-pair engineering.


Assuntos
Peroxidase , Zinco , Humanos , Catálise , Peroxidase/metabolismo , Peroxidase/química , Zinco/química , Zinco/metabolismo , Animais , Domínio Catalítico , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Camundongos , Linhagem Celular Tumoral , Teoria da Densidade Funcional
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