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1.
Biomed Pharmacother ; 167: 115497, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37741253

RESUMEN

Sepsis-induced myocardial dysfunction (SIMD) is one of the serious health-affecting problems worldwide. At present, the mechanisms of SIMD are still not clearly elucidated. The NOD-like receptor protein 3 (NLRP3) inflammasome has been assumed to be involved in the pathophysiology of SIMD by regulating multiple biological processes. NLRP3 inflammasome and its related signaling pathways might affect the regulation of inflammation, autophagy, apoptosis, and pyroptosis in SIMD. A few molecular specific inhibitors of NLRP3 inflammasome (e.g., Melatonin, Ulinastatin, Irisin, Nifuroxazide, and Ginsenoside Rg1, etc.) have been developed, which showed a promising anti-inflammatory effect in a cellular or animal model of SIMD. These experimental findings indicated that NLRP3 inflammasome could be a promising therapeutic target for SIMD treatment. However, the clinical translation of NLRP3 inhibitors for treating SIMD still requires robust in vivo and preclinical trials.

2.
J Fluoresc ; 32(1): 397-404, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34977993

RESUMEN

Reported here is a new [Cu4I4] cluster-based coordination polymer, namely [Cu4I4(bib)2]n·n(DMF) (1, bib = 1,4-bis(imidazolyl)butane, DMF = N,N'-dimethylformamide), which was synthesized by the self-assemble reaction of CuI, bib and KI under solvothermal conditions. Remarkably, compound 1 shows promising photocatalytic performance toward to the degradation of MB solution under visible light irradiation. For the COPD treatment, the ELISA detection kit was conducted to determine the content of INF-γ released by the respiratory tract mucosal epithelial cells. In addition to this, the activation levels of the NF-κB signaling pathway were still need to be assessed by the real time RT-PCR after the compound treatment.


Asunto(s)
Cobre/química , Cobre/farmacología , Interferón gamma/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Animales , Catálisis , Células Epiteliales/metabolismo , Humanos , Dosificación Letal Mediana , Ratones , FN-kappa B/metabolismo , Procesos Fotoquímicos , Polímeros , Reacción en Cadena en Tiempo Real de la Polimerasa , Mucosa Respiratoria/citología , Mucosa Respiratoria/metabolismo , Transducción de Señal , Difracción de Rayos X
3.
Mol Med Rep ; 19(2): 1331-1339, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30535506

RESUMEN

Epidermal growth factor receptor­tyrosine kinase inhibitors, such as gefitinib, have been found to be clinically effective in the treatment of patients with non­small cell lung cancer (NSCLC). However, the therapeutic effect of gefitinib is often limited by the development of gefitinib resistance. MicroRNAs (miRNAs), a group of small non­coding RNAs, have been demonstrated to be frequently dysregulated in human malignancies. For instance, the downregulation of miR­506­3p has been reported in NSCLC patients. The aim of the present study was to determine the role and underlying molecular mechanism of miR­506­3p in the regulation of gefitinib sensitivity in NSCLC. A gefitinib­resistant PC­9 (PC­9GR) cell line was established, and reduced miR­506­3p expression was observed in PC­9GR cells as compared with that in parental cells. The results of cell cytotoxicity and cell apoptosis assays indicated that PC­9GR cells were more sensitive to gefitinib following the transfection with an miR­506­3p mimic, while transfection with an miR­506­3p antagonist reduced the sensitivity of PC­9GR cells to gefitinib. It was further revealed that Yes­associated protein 1 (YAP1) was directly suppressed by miR­506­3p in PC­9GR cells. The elevated sensitivity of PC­9GR cells to gefitinib following transfection with the miR­506­3p mimic was counteracted by the overexpression of YAP1. Furthermore, an inverse correlation between the miR­506­3p and YAP1 mRNA levels was detected in lung adenocarcinoma specimens. Collectively, the results of the present study suggested that the downregulation of miR­506­3p contributes to gefitinib resistance, and thus, the restoration of miR­506­3p may be a potential therapeutic approach for overcoming NSCLC gefitinib resistance.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Resistencia a Antineoplásicos/genética , Neoplasias Pulmonares/genética , MicroARNs/genética , Fosfoproteínas/genética , Adenocarcinoma del Pulmón/tratamiento farmacológico , Adenocarcinoma del Pulmón/genética , Anciano , Antineoplásicos/uso terapéutico , Apoptosis/genética , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Línea Celular , Línea Celular Tumoral , Regulación hacia Abajo/genética , Femenino , Gefitinib/uso terapéutico , Células HEK293 , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Masculino , Persona de Mediana Edad , ARN Mensajero/genética , Factores de Transcripción , Proteínas Señalizadoras YAP
4.
Cancer Biother Radiopharm ; 32(4): 115-123, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28514208

RESUMEN

BACKGROUND: MicroRNA-516b (miR-516b) has been recently reported to be downregulated in nonsmall cell lung cancer (NSCLC). However, its clinical significance and biological function in NSCLC remain to be clarified. MATERIALS AND METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-516b in 82 paired fresh primary tumor tissues and NSCLC cell lines. The association of miR-516b expression with clinicopathological factors and prognosis was statistically analyzed by SPSS 21.0 software, Kaplan-Meier method, and Cox regression analyses. Cell Counting Kit-8, colony formation, flow cytometric, Transwell migration, and invasion assays were used to evaluate the proliferation, cell cycle, apoptosis, migration, and invasion of NSCLC cells after miR-516b mimics or negative control of mimics transfection. RESULTS: The expression level of miR-516b was found to be significantly lower in NSCLC tissues and cell lines than in corresponding normal tissues and cells. Decreased miR-516b expression was significantly associated with tumor size (p = 0.004), Tumor Node Metastasis (TNM) stage (p = 0.016), and shorter overall survival (p = 0.0039). Multivariate analysis suggested that miR-516b was an independent risk factor for NSCLC (hazard ratio = 2.435, 95% confidence interval: 1.423-2.457; p = 0.003). Furthermore, overexpression of miR-516b could inhibit NSCLC cell proliferation, cell cycle progression, migration and invasion, and promoted cell apoptosis. The qRT-PCR results indicated that overexpressing miR-516b reduced the mRNA expression of CDK2, MMP-2, and MMP-9, whereas increased BAX mRNA expression in NSCLC cells. Their protein expression levels presented similar trends, as confirmed by Western blotting. CONCLUSIONS: Findings in this study demonstrated for the first time that miR-516b expression might be a novel diagnostic and prognostic factor, as well as a promising target for NSCLC.


Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/genética , Neoplasias Pulmonares/genética , MicroARNs/biosíntesis , Células A549 , Apoptosis/genética , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Ciclo Celular/genética , Línea Celular Tumoral , Femenino , Genes Supresores de Tumor , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Masculino , MicroARNs/genética , MicroARNs/metabolismo , Persona de Mediana Edad , Estadificación de Neoplasias , Pronóstico , Transfección
5.
Talanta ; 107: 176-82, 2013 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-23598209

RESUMEN

A novel electrochemical immunosensor for sensitive detection of clenbuterol (CLB) is fabricated using glucose oxidase (GOD)-functionalized grahene oxide (GO) nanocomposites to label CLB. The immunosensor was constructed by layer-by-layer assembly colloidal prussian blue (PB), multiwalled carbon nanotubes (MWCNTs) and CLB antibodies (Abs) on a glassy carbon electrode (GCE). In this competitive immunoassay system, PB acts as the redox mediator to reduce H2O2 originated from the catalyst cycle of GOD. The high ratio of GOD to GO effectively amplified the signal for this competitive-type immunoassay. Under optimized conditions, the immunosensor shows a wide linear range from 0.5 to 1,000 ng/mL with a low detection limit of 0.25 ng/mL. The dual signal amplification of GOD-functionalized GO nanocomposites as a label is promising to be applied to design other sensitive immunosenseors.


Asunto(s)
Agonistas Adrenérgicos beta/análisis , Alimentación Animal/análisis , Clenbuterol/análisis , Grafito/química , Inmunoensayo/métodos , Óxidos/química , Animales , Aspergillus niger/enzimología , Técnicas Electroquímicas/métodos , Enzimas Inmovilizadas/química , Glucosa Oxidasa/química , Oro/química , Límite de Detección , Nanopartículas/química , Porcinos
6.
Biosens Bioelectron ; 45: 292-9, 2013 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-23500378

RESUMEN

In this report, a novel visible-light-activated photoelectrochemical biosensor was fabricated to study the inhibition of acetylcholinesterase (AChE) activity induced by two endogenous neurotoxins, 1(R)-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline [(R)-Sal] and 1(R),2(N)-dimethyl-6,7-dihydroxy-1,2,3,4-tetra-hydroisoquinoline [(R)-NMSal], which have drawn much attention in the study of the pathogenesis of neurodegenerative diseases such as Parkinson's disease. The photoelectrode was prepared by three steps, as follows. At first, nitrogen and fluorine co-doped TiO2 nanotubes (TNs) were obtained by anodic oxidation of a Ti sheet. Secondly, silver nanoparticles (AgNPs) were deposited onto the TNs through a microwave-assisted heating polyol (MAHP) process. At last, AChE was immobilized on the obtained photoelectrode and the biosensor was marked as AChE/Ag/NFTNs. Due to the nitrogen and fluorine co-doping, the photoelectrochemical biosensors can produce high photocurrent under visible light irradiation. Moreover, the presence of AgNPs greatly increased the photocurrent response of the biosensor. AChE/Ag/NFTNs hybrid system was used to study AChE inhibition induced by (R)-Sal and (R)-NMSal. The result proved that both (R)-Sal and (R)-NMSal exhibited mixed and reversible inhibition against AChE. This strategy is of great significance for the development of novel photoelectrochemical biosensors in the future.


Asunto(s)
Acetilcolinesterasa/aislamiento & purificación , Acetilcolinesterasa/metabolismo , Técnicas Biosensibles , Enfermedad de Parkinson/enzimología , Acetilcolinesterasa/química , Inhibidores de la Colinesterasa/farmacología , Electroquímica , Luz , Nanotubos/química , Neurotoxinas/farmacología , Tetrahidroisoquinolinas/farmacología
7.
J Phys Chem B ; 116(36): 11180-6, 2012 Sep 13.
Artículo en Inglés | MEDLINE | ID: mdl-22946763

RESUMEN

In this paper, an electrochemical acetylcholinesterase (AChE) inhibition assay based on three-dimensional ordered macroporous (3DOM) composite was conducted. The 3DOM composite was first fabricated on the glassy carbon electrode by electropolymerization of aniline in the presence of ionic liquid (IL) on a sacrificial silica nanospheres template. After the silica nanospheres were etched, an IL-doped polyaniline (IL-PANI) film with 3DOM morphology was formed. Then, gold nanoparticles (AuNPs) were decorated on the IL-PANI film by electrodeposition. The immobilized AChE on the 3DOM composite displayed favorable affinity to substrate acetylthiocholine chloride (ATCh), and the 3DOM composite showed excellent electrocatalytic effect on thiocholine, the hydrolysis product of ATCh. The presence of IL and AuNPs could improve the sensitivity by accelerating the electron transfer. The designed AChE biosensor was successfully applied to evaluate the AChE inhibition induced by endogenous neurotoxin 1(R),2N-dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline [(R)-NMSal]. The results demonstrate that (R)-NMSal exerts a considerable effect on AChE activity, and the inhibition is reversible. The developed method offers a new approach for AChE inhibition assay, which is of great benefit in understanding the mechanism behind neurotoxin-induced neurodegenerative disorders.


Asunto(s)
Acetilcolinesterasa/metabolismo , Inhibidores de la Colinesterasa/farmacología , Electrophorus/metabolismo , Enzimas Inmovilizadas/metabolismo , Nanocompuestos/química , Tetrahidroisoquinolinas/farmacología , Compuestos de Anilina/química , Animales , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Porosidad
8.
Analyst ; 137(18): 4349-55, 2012 Sep 21.
Artículo en Inglés | MEDLINE | ID: mdl-22830075

RESUMEN

We report the development of an ultrasensitive amperometric biosensor based on Ag nanoparticles-decorated graphene oxide nanosheets (GO) (Ag-GO) for the rapid detection of clenbuterol (CLB). The morphology and structure of the Ag-GO labeled CLB (Ag-GO-CLB) were characterized by transmission electron microscope (TEM), atomic force microscope (AFM), and ultraviolet-visible spectroscope (UV-vis). The immunosensor was prepared by covalently immobilizing capture antibodies on a multi-walled carbon nanotubes-modified glassy carbon electrode. Through competitive immunoreactions, the Ag-GO-CLB nanocomposites were captured on the immunosensor and the silver was measured by positive differential pulse voltammetry (DPV) in KCl solution for the detection of antigen. The experimental results show a linear response over the range from 0.01 to 10.0 ng mL(-1) with a lower detection limit of 6.8 pg mL(-1) (signal-to-noise ratio of 3). The Ag-GO based immunosensor offers a simple and convenient route for metal-immunoassay labels, which can avoid the complicated and time-consuming dissolving of metal component for ultrasensitive determination. Moreover, the electrochemical immunoassay shows acceptable specificity and stability and is suitable for the determination of CLB in real samples.

9.
Biosens Bioelectron ; 32(1): 224-30, 2012 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-22221797

RESUMEN

A novel electrochemical immunosensor for sensitive detection of α-synuclein (α-SYN), a very important neuronal protein, has been developed based on dual signal amplification strategy. Herein, G4-polyamidoamine dendrimer-encapsulated Au nanoparticles (PAMAM-Au nanocomposites) were covalently bound on the poly-o-aminobenzoic acid (poly-o-ABA), which was initially electropolymerized on the electrode surface to perform abundant carboxyl groups. The formed immunosensor platform, PAMAM-Au, was proved to provide numerous amino groups to allow highly dense immobilization of antigen, and facilitate the improvement of electrochemical responses as well. Subsequently, the enhanced gold nanoparticle labels ({HRP-Ab(2)-GNPs}) were fabricated by immobilizing horseradish peroxidase-secondary antibody (HRP-Ab(2)) on the surface of gold nanoparticles (GNPs). After an immunoassay process, the {HRP-Ab(2)-GNPs} labels were introduced onto the electrode surface, and produced an electrocatalytic response by reduction of hydrogen peroxide (H(2)O(2)) in the presence of enzymatically oxidized thionine. On the basis of the dual signal amplification of PAMAM-Au and {HRP-Ab(2)-GNPs} labels, the designed immunosensor displayed an excellent analytical performance with high sensitivity and stability. This developed strategy was successfully proved as a simple, cost-effective method, and could be easily extended to other protein analysis schemes.


Asunto(s)
Dendrímeros/química , Técnicas Electroquímicas/métodos , Técnicas Electroquímicas/estadística & datos numéricos , Oro/química , Inmunoensayo/métodos , Nanopartículas/química , alfa-Sinucleína/análisis , Anticuerpos Inmovilizados/inmunología , Humanos , Sensibilidad y Especificidad , alfa-Sinucleína/inmunología
10.
J Environ Sci (China) ; 24(9): 1717-22, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-23520882

RESUMEN

A simple and sensitive electroanalytical method was presented for the determination of 4-n-octylphenol (OP) based on multi-walled carbon nanotubes (MWCNTs) modified glassy carbon electrode (GCE). OP was directly oxidized on the MWCNTs/GCE, and the electrochemical oxidation mechanism was demonstrated by a one-electron and one-proton process in the reaction. The oxidation peak current of OP was significantly enhanced by the use of MWCNTs/GCE compared with those of bare glassy carbon electrode, suggesting that the modified electrode can remarkably improve the performance for OP determination. Factors influencing the detection processes were optimized. Under these optimal conditions, a linear relationship between concentration of OP and current response was obtained in the range of 5 x 10(-8) to 1 x 10(-5) mol/L with a detection limit of 1.5 x 10(-8) mol/L and correlation coefficient 0.9986. The modified electrode showed good selectivity, sensitivity, reproducibility and high stability.


Asunto(s)
Nanotubos de Carbono/química , Fenoles/química , Electrodos , Monitoreo del Ambiente
11.
Talanta ; 87: 100-5, 2011 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-22099655

RESUMEN

A novel label-free electrochemical immunosensor for rapid determination of chloramphenicol (CAP) was fabricated by entrapping monoclonal antibody to chloramphenicol (anti-CAP) in hollow gold nanospheres (HGNs)/chitosan composite modified on a glassy carbon electrode. The hollow gold nanospheres (HGNs) were prepared by using Co nanoparticles as sacrificial templates and characterized by transmission electron microscopy (TEM). The changes of the electrode behavior after each fabrication step were investigated by electrochemical impedance spectroscopy (EIS) technique. Under optimal conditions, the proposed immunosensor has a sensitive response to CAP in a linear range of 0.1-1000 ng mL(-1) with the detection limit of 0.06 ng mL(-1). Accurate detection of CAP in real meat samples was demonstrated by comparison with conventional HPLC method. The proposed method was proven to be a feasible quantitative method for CAP analysis with the properties of simple preparation, stability, high sensitivity and selectivity.


Asunto(s)
Antibacterianos/análisis , Anticuerpos Inmovilizados/inmunología , Cloranfenicol/análisis , Inmunoensayo/métodos , Carne/análisis , Nanosferas/química , Animales , Antibacterianos/inmunología , Anticuerpos Inmovilizados/química , Bovinos , Quitosano/química , Cloranfenicol/inmunología , Técnicas Electroquímicas/métodos , Productos Pesqueros/análisis , Peces , Oro/química , Nanosferas/ultraestructura , Sensibilidad y Especificidad , Porcinos
12.
Biosens Bioelectron ; 26(12): 4661-6, 2011 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-21733671

RESUMEN

A novel amplified electrochemical immunoassay based on ferrocene (Fc)-functionalized ZnO nanorods (NRs) was developed in the present work. The detection antibody ((d)Ab) and Fc were immobilized onto the surface of ZnO NRs, denoted as {(d)Ab-ZnO-Fc} bioconjugates. The amount of (d)Ab and Fc in the bioconjugates was investigated using the copper reduction/bicinchoninic acid reaction (BCA protein assay) and inductive coupled plasma-atomic emission spectroscopy (ICP-AES), respectively. Greatly amplified signal was achieved in the sandwich-type immunoassay when (d)Ab and Fc linked to ZnO NRs at a proper ratio. Using Escherichia coli (E. coli) as a model antigen, the designed immunoassay showed an excellent analytical performance, and exhibited a wide dynamic response range of E. coli concentration from 10(2) to 10(6)cfu/mL with a detection limit of 50 cfu/mL (S/N=3). By introducing a pre-enrichment step, the detection of 5 cfu/10 mL E. coli in hospital sewage water was realized. This proposed signal amplification strategy was promising and could be easily extended to monitor other biorecognition events.


Asunto(s)
Anticuerpos Inmovilizados/química , Técnicas Electroquímicas/métodos , Escherichia coli/aislamiento & purificación , Compuestos Ferrosos/química , Nanotubos/química , Anticuerpos Inmovilizados/inmunología , Técnicas Biosensibles/métodos , Escherichia coli/inmunología , Inmunoensayo/métodos , Límite de Detección , Metalocenos , Nanotubos/ultraestructura , Óxido de Zinc/química
13.
Anal Chim Acta ; 697(1-2): 27-31, 2011 Jul 04.
Artículo en Inglés | MEDLINE | ID: mdl-21641415

RESUMEN

In this paper, a simple two-step approach for redox protein immobilization was introduced. Firstly, alkynyl-terminated film was formed on electrode surface by electrochemical reduction of 4-ethylnylphenyl (4-EP) diazonium compound. Then, horseradish peroxidase (HRP) modified with azido group was covalently immobilized onto the electrografted film via click reaction. Reflection absorption infrared (RAIR) spectroscopy and electrochemical methods were used to characterize the modification process. The results indicate that HRP retains its native structure and shows fast direct electron transfer. Moreover, the immobilized HRP shows excellent electrocatalytic reduction activity toward H(2)O(2) with a linear range of 5.0×10(-6) to 9.3×10(-4) mol L(-1).


Asunto(s)
Química Clic , Compuestos de Diazonio/química , Electroquímica/métodos , Enzimas Inmovilizadas/química , Peroxidasa de Rábano Silvestre/química , Adsorción , Biocatálisis , Electrodos , Transporte de Electrón , Enzimas Inmovilizadas/metabolismo , Oro/química , Peroxidasa de Rábano Silvestre/metabolismo , Peróxido de Hidrógeno/análisis , Peróxido de Hidrógeno/metabolismo , Modelos Moleculares , Conformación Proteica
14.
Talanta ; 84(3): 607-13, 2011 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-21482257

RESUMEN

A Fe(2)O(3)@Au core/shell nanoparticle-based electrochemical DNA biosensor was developed for the amperometric detection of Escherichia coli (E. coli). Magnetic Fe(2)O(3)@Au nanoparticles were prepared by reducing HAuCl(4) on the surfaces of Fe(2)O(3) nanoparticles. This DNA biosensor is based on a sandwich detection strategy, which involves capture probe immobilized on magnetic nanoparticles (MNPs), target and reporter probe labeled with horseradish peroxidase (HRP). Once magnetic field was added, these sandwich complexes were magnetically separated and HRP confined at the surfaces of MNPs could catalyze the enzyme substrate and generate electrochemical signals. The biosensor could detect the concentrations upper than 0.01 pM DNA target and upper than 500 cfu/mL of E. coli without any nucleic acid amplification steps. The detection limit could be lowered to 5 cfu/mL of E. coli after 4.0 h of incubation.


Asunto(s)
Técnicas Biosensibles , ADN/química , Electroquímica/instrumentación , Escherichia coli/aislamiento & purificación , Compuestos Férricos/química , Nanopartículas del Metal , Secuencia de Bases , Recuento de Colonia Microbiana , Sondas de ADN , Enzimas Inmovilizadas/química , Peroxidasa de Rábano Silvestre/química , Límite de Detección
15.
Bioelectrochemistry ; 81(1): 53-7, 2011 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-21277265

RESUMEN

In this work, an amine-terminated poly (amidoamine) dendrimer containing Pt nanoparticles (PAMAM/Pt) nanocomposite was synthesized and a novel amperometric H(2)O(2) biosensor based on PAMAM/Pt and MWCNTs was developed. The resulting film of MWCNTs/PAMAM/Pt was characterized by transmission electron microscopy (TEM), linear sweep voltammetry (LSV) and amperometric i-t curve. It demonstrates excellent electrocatalytic responses toward the reduction of H(2)O(2) at -200 mV (vs.SCE) without HRP participation. Immobilized with glutamate oxidase (GlutaOx), an effective glutamate biosensor, was fabricated, and the in vivo detection for glutamate was realized combining with the on-line microdialysis system. The glutamate biosensor showed good linear range from 1.0 µM to 50.0 µM with the detection limit of 0.5 µM (S/N=3). The basal level of glutamate in the striatum of rat was detected continuously with this on-line system and was calculated to be 5.80±0.12 µM (n=3). This method was proved to be sensitive and selective and may be feasible in the further application of physiology and pathology.


Asunto(s)
Técnicas Biosensibles/métodos , Dendrímeros/química , Ácido Glutámico/análisis , Microdiálisis/métodos , Platino (Metal)/química , Animales , Técnicas Biosensibles/instrumentación , Química Encefálica , Conductividad Eléctrica , Electroquímica , Peróxido de Hidrógeno/química , Masculino , Microdiálisis/instrumentación , Nanopartículas , Ratas , Ratas Sprague-Dawley
16.
Biosens Bioelectron ; 26(7): 3325-30, 2011 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-21277764

RESUMEN

A new type of DNA sequence-specific electrochemical biosensor based on magnetic beads for the detection of Escherichia coli is reported in the present work. Alginic acid-coated cobalt magnetic beads, capped with 5'-(NH(2)) oligonucleotide and employed not only for magnetic separation but also as the solid adsorbent, were used as DNA probes to hybridize with the target E. coli DNA sequence. This assay was specific for E. coli detection depending on the uid A gene, which encodes for the enzyme ß-d-glucuronidase produced by E. coli strains. When daunomycin (DNR) was used as DNA hybridization indicator, the target sequences of E. coli hybridized with the probes resulted in the decrease of DNR reduction peak current, which was proportional to the E. coli concentration. The optimization of the hybridization detection was carried out and the specificity of the probes was also demonstrated. This DNA biosensor can be employed to detect a complementary target sequence for 3.0×10(-10) mol/L and denatured PCR products for 0.5 ng/µL. The linear range of the developed biosensor for the detection of E. coli cells was from 1.0×10(2) to 2.0×10(3) cells/mL with a detection limit of 50 cells/mL. After a brief enrichment process, a concentration of 10 cells/mL E. coli in real water samples was detected by the electrochemical biosensor.


Asunto(s)
Alginatos/química , Técnicas Biosensibles/métodos , Cobalto/química , Técnicas Electroquímicas/métodos , Escherichia coli/aislamiento & purificación , Microbiología del Agua , Secuencia de Bases , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Escherichia coli/genética , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Magnetismo , Hibridación de Ácido Nucleico , Sensibilidad y Especificidad
17.
Talanta ; 84(1): 204-11, 2011 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-21315921

RESUMEN

Sudan I monoclonal antibodies (Mabs) were prepared by hybridoma technique and firstly used to develop a Sudan I immunosensor by immobilizing the Mabs on a gold electrode. o-Mercaptobenzoic acid (MBA) was covalently conjugated on the gold electrode to form a self-assembled monolayer (SAM). The immobilization of Sudan I Mabs to the SAM was carried out through a stable acyl amino ester intermediate generated by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydrosuccinimide (NHS), which could condense antibodies reproducibly and densely on the SAM. The changes of the electrode behavior after each assembly step were investigated by cyclic voltammetric (CV) technique. The Sudan I concentration was measured through the increase of impedance values in the corresponding specific binding of Sudan I and Sudan I antibody. A linear relationship between the increased electron-transfer resistance (Ret) and the logarithmic value of Sudan I concentrations was found in the range of 0.05-50 ng mL(-1) with the detection limit of 0.03 ng mL(-1). Using hot chili as a model sample, acceptable recovery of 96.5-107.3% was obtained. The results were validated by conventional HPLC method with good correlation. The proposed method was proven to be a feasible quantitative method for Sudan I analysis with the properties of stability, highly sensitivity and selectivity.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Técnicas Biosensibles/métodos , Espectroscopía Dieléctrica/métodos , Inmunoensayo/métodos , Naftoles/análisis , Animales , Anticuerpos Inmovilizados/química , Anticuerpos Inmovilizados/inmunología , Anticuerpos Monoclonales/química , Técnicas Biosensibles/economía , Calibración , Espectroscopía Dieléctrica/economía , Electroquímica , Electrodos , Transporte de Electrón , Ésteres , Etildimetilaminopropil Carbodiimida/química , Femenino , Análisis de los Alimentos , Oro/química , Inmunoensayo/economía , Límite de Detección , Modelos Lineales , Ratones , Naftoles/inmunología , Succinimidas/química , Propiedades de Superficie
18.
Talanta ; 83(5): 1381-5, 2011 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-21238725

RESUMEN

A simple and versatile approach for covalent immobilization of redox protein on solid surface via self-assembled technique and click chemistry is reported. The alkynyl-terminated monolayers are obtained by self-assembled technique, then, azido-horseradish peroxidase (azido-HRP) was covalent immobilized onto the formed monolayers by click reaction. The modified process is characterized by reflection absorption infrared spectroscopy (RAIR), surface-enhanced Raman scattering spectroscopy (SERS) and electrochemical methods. All the experimental results suggest that HRP is immobilized onto the electrode surface successfully without denaturation. Furthermore, the immobilized HRP shows electrocatalytic reduction for H(2)O(2), and the linear range is from 5.0 to 700 µM. The heterogeneous electron transfer rate constant k(s) is 1.11 s(-1) and the apparent Michaelis-Menten constant is calculated to be 0.196 mM.


Asunto(s)
Enzimas Inmovilizadas/química , Peroxidasa de Rábano Silvestre/química , Técnicas Biosensibles/tendencias , Electroquímica , Espectroscopía Infrarroja Corta
19.
Biosens Bioelectron ; 26(7): 3227-32, 2011 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-21239160

RESUMEN

A new type of hydroxyl functionalized room temperature ionic liquid (RTIL), [C(3)(OH)(2)mim][BF(4)], was synthesized herein and a novel H(2)O(2) biosensor is fabricated with [C(3)(OH)(2)mim][BF(4)] as the substrate and electrodepositing bimetallic Au/Pt nanoparticles (NPs) onto the [C(3)(OH)(2)mim][BF(4)] film. The functionalization of RTIL with hydroxyl groups provided an appropriate environment for the preparation of more uniform and smaller Au/Pt NPs with the diameter of 2.5 nm±0.2 nm. Immobilized with glutamate oxidase (GlutaOx), the resulting GlutaOx-[C(3)(OH)(2)mim][BF(4)]-Au/Pt-Nafion biosensor displayed excellent electrocatalytic response to glutamate at a potential of -200 mV. An effective on-line microdialysis system, which was powered by a microdialysis pump, was set up and used for the detection of glutamate successively in the striatum of rats. The glutamate biosensor in on-line microdialysis system showed good linear range from 0.5 µM to 20.0 µM with the detection limit of 0.17 µM (S/N=3). The basal level of glutamate in the striatum of anaesthetic rats was calculated to be 3.01±0.67 µM (n=3). The application of the GlutaOx-[C(3)(OH)(2)mim][BF(4)]-Au/Pt-Nafion electrode is further demonstrated for in vivo sensing of the variation of glutamate level in the striatum when rats received intraperitoneal (i.p.) injection of 100 mM KCl and brain electrical stimulation of the subthalamic nucleus area (STN). Both of the two kinds of stimulation resulted in an increase in the extracellular concentration of glutamate. This method has proved to be sensitive and reproducible, which enables its promising application in physiology and pathology.


Asunto(s)
Técnicas Biosensibles/instrumentación , Cuerpo Estriado/metabolismo , Ácido Glutámico/análisis , Líquidos Iónicos/química , Microdiálisis/instrumentación , Aminoácido Oxidorreductasas/metabolismo , Animales , Electrodos , Enzimas Inmovilizadas/metabolismo , Diseño de Equipo , Ácido Glutámico/metabolismo , Oro/química , Peróxido de Hidrógeno/metabolismo , Nanopartículas del Metal/química , Nanopartículas del Metal/ultraestructura , Platino (Metal)/química , Ratas , Sensibilidad y Especificidad
20.
Chemistry ; 16(48): 14439-46, 2010 Dec 27.
Artículo en Inglés | MEDLINE | ID: mdl-21038326

RESUMEN

α-Synuclein (α-SYN) is a very important neuronal protein that is associated with Parkinson's disease. In this paper, we utilized Au-doped TiO(2) nanotube arrays to design a photoelectrochemical immunosensor for the detection of α-SYN. The highly ordered TiO(2) nanotubes were fabricated by using an electrochemical anodization technique on pure Ti foil. After that, a photoelectrochemical deposition method was exploited to modify the resulting nanotubes with Au nanoparticles, which have been demonstrated to facilitate the improvement of photocurrent responses. Moreover, the Au-doped TiO(2) nanotubes formed effective antibody immobilization arrays and immobilized primary antibodies (Ab(1)) with high stability and bioactivity to bind target α-SYN. The enhanced sensitivity was obtained by using {Ab(2)-Au-GOx} bioconjugates, which featured secondary antibody (Ab(2)) and glucose oxidase (GOx) labels linked to Au nanoparticles for signal amplification. The GOx enzyme immobilized on the prepared immunosensor could catalyze glucose in the detection solution to produce H(2)O(2), which acted as a sacrificial electron donor to scavenge the photogenerated holes in the valence band of TiO(2) nanotubes upon irradiation of the other side of the Ti foil and led to a prompt photocurrent. The photocurrents were proportional to the α-SYN concentrations, and the linear range of the developed immunosensor was from 50 pg mL(-1) to 100 ng mL(-1) with a detection limit of 34 pg mL(-1). The proposed method showed high sensitivity, stability, reproducibility, and could become a promising technique for protein detection.


Asunto(s)
Anticuerpos Inmovilizados/análisis , Compuestos de Boro , Oro/química , Nanopartículas del Metal , Titanio , alfa-Sinucleína/análisis , Aspergillus niger/enzimología , Técnicas Electroquímicas , Glucosa Oxidasa/metabolismo , Humanos , Inmunoensayo , Nanotubos , Enfermedad de Parkinson/patología
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