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1.
Talanta ; 234: 122672, 2021 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-34364473

RESUMEN

An Enzyme Linked ImmunoMagnetic Electrochemical assay (ELIME) was developed for the detection of the hepatitis A virus (HAV). This system is based on the use of new polydopamine-modified magnetic nanobeads as solid support for the immunochemical chain, and an array of 8 screen-printed electrodes as a sensing platform. Enzymatic-by-product is quickly measured by differential pulse voltammetry. For this purpose, all analytical parameters were optimized; in particular, different blocking reagents were evaluated in order to minimize the nonspecific interaction of bioreagents. Using the ELIME assays, a quantitative determination of HAV can be achieved with a detection limit of 1·10-11 IU mL-1 and a working range between 10-10 - 5 × 10-7 IU mL-1. The cross-reactivity of the commercial monoclonal antibodies against HAV used in ELIME assays was tested for Coxsackie B4, resulting very low. The sensitivity was also investigated and compared with spectrophotometric sandwich ELISA. The average relative standard deviation (RSD) of the ELIME method was less than 5% for the assays performed on the same day, and 7% for the measurements made on different days. The proposed system was applied to the cell culture of HAV, which title was quantified by Real-Time Quantitative Reverse Transcription PCR (RT¬qPCR). To compare the results, a correlation between the units used in ELIME (IU mL-1) and those used in RT¬qPCR (genome mL-1) was established using a HAV-positive sample, resulting in 1 IU mL-1-10-4 gen mL-1 (R2 = 0.978). The ELIME tool exhibits good stability and high biological selectivity for HAV antigen detection and was successfully applied for the determination of HAV in tap water.


Asunto(s)
Virus de la Hepatitis A , Bioensayo , Virus de la Hepatitis A/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sensibilidad y Especificidad
3.
Anal Chem ; 92(9): 6654-6666, 2020 05 05.
Artículo en Inglés | MEDLINE | ID: mdl-32252524

RESUMEN

DJ-1, a 20.7 kDa protein, is overexpressed in people who have bladder cancer (BC). Its elevated concentration in urine allows it to serve as a marker for BC. However, no biosensor for the detection of DJ-1 has been demonstrated. Here, we describe a virus bioresistor (VBR) capable of detecting DJ-1 in urine at a concentration of 10 pM in 1 min. The VBR consists of a pair of millimeter-scale gold electrodes that measure the electrical impedance of an ultrathin (≈ 150-200 nm), two-layer polymeric channel. The top layer of this channel (90-105 nm in thickness) consists of an electrodeposited virus-PEDOT (PEDOT is poly(3,4-ethylenedioxythiophene)) composite containing embedded M13 virus particles that are engineered to recognize and bind to the target protein of interest, DJ-1. The bottom layer consists of spin-coated PEDOT-PSS (poly(styrenesulfonate)). Together, these two layers constitute a current divider. We demonstrate here that reducing the thickness of the bottom PEDOT-PSS layer increases its resistance and concentrates the resistance drop of the channel in the top virus-PEDOT layer, thereby increasing the sensitivity of the VBR and enabling the detection of DJ-1. Large signal amplitudes coupled with the inherent simplicity of the VBR sensor design result in high signal-to-noise (S/N > 100) and excellent sensor-to-sensor reproducibility characterized by coefficients of variation in the range of 3-7% across the DJ-1 binding curve down to a concentration of 30 pM, near the 10 pM limit of detection (LOD), encompassing four orders of magnitude in concentration.


Asunto(s)
Bacteriófago M13/química , Biomarcadores de Tumor/orina , Técnicas Biosensibles , Proteína Desglicasa DJ-1/orina , Neoplasias de la Vejiga Urinaria/orina , Humanos , Factores de Tiempo
4.
Biomed Res Int ; 2019: 4568039, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31781615

RESUMEN

Chemical structures derived from marine foods are highly diverse and pharmacologically promising. In particular, chitooligosaccharides (COS) present a safe pharmacokinetic profile and a great source of new bioactive polymers. This review describes the antioxidant, anti-inflammatory, and antidiabetic properties of COS from recent publications. Thus, COS constitute an effective agent against oxidative stress, cellular damage, and inflammatory pathogenesis. The mechanisms of action and targeted therapeutic pathways of COS are summarized and discussed. COS may act as antioxidants via their radical scavenging activity and by decreasing oxidative stress markers. The mechanism of COS antidiabetic effect is characterized by an acceleration of pancreatic islets proliferation, an increase in insulin secretion and sensitivity, a reduction of postprandial glucose, and an improvement of glucose uptake. COS upregulate the GLUT2 and inhibit digestive enzyme and glucose transporters. Furthermore, they resulted in reduction of gluconeogenesis and promotion of glucose conversion. On the other hand, the COS decrease inflammatory mediators, suppress the activation of NF-κB, increase the phosphorylation of kinase, and stimulate the proliferation of lymphocytes. Overall, this review brings evidence from experimental data about protective effect of COS.


Asunto(s)
Antiinflamatorios , Quitina/análogos & derivados , Depuradores de Radicales Libres , Hipoglucemiantes , Animales , Antiinflamatorios/farmacocinética , Antiinflamatorios/uso terapéutico , Quitina/farmacocinética , Quitina/uso terapéutico , Quitosano , Depuradores de Radicales Libres/farmacocinética , Depuradores de Radicales Libres/uso terapéutico , Gluconeogénesis/efectos de los fármacos , Glucosa/metabolismo , Transportador de Glucosa de Tipo 2/metabolismo , Humanos , Hipoglucemiantes/farmacocinética , Hipoglucemiantes/uso terapéutico , Oligosacáridos
5.
ACS Appl Mater Interfaces ; 11(5): 4757-4765, 2019 Feb 06.
Artículo en Inglés | MEDLINE | ID: mdl-30668098

RESUMEN

A polymer-based electrode capable of specific detection of human serum albumin, and its glycated derivatives, is described. The sensor is constructed from a glass microscope slide coated with a synthesized, polythiophene film bearing a protected, iminodiacetic acid motif. The electrode surface is then further elaborated to a functional biosensor through deprotection of the iminodiacetic acid, followed by metal-affinity immobilization of a specific and high-affinity, albumin ligand. Albumin was then quantified in buffer and synthetic urine via electrochemical impedance spectroscopy. Glycated albumin was next bound to a boronic acid-modified, single-cysteine dihydrofolate reductase variant to quantify glycation ratios by square-wave voltammetry. The platform offers high sensitivity, specificity, and reproducibility in an inexpensive arrangement. The detection limits exceed the requirements for intermediate-term glycemic control monitoring in diabetes patients at 5 and 1 nM for albumin and its glycated forms, respectively.


Asunto(s)
Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Albúmina Sérica Humana/orina , Albúmina Sérica/análisis , Enzimas Inmovilizadas/química , Enzimas Inmovilizadas/metabolismo , Diseño de Equipo , Productos Finales de Glicación Avanzada , Humanos , Modelos Biológicos , Tetrahidrofolato Deshidrogenasa/química , Tetrahidrofolato Deshidrogenasa/metabolismo , Albúmina Sérica Glicada
6.
Rev. bras. farmacogn ; 28(5): 575-581, Sept.-Oct. 2018. tab, graf
Artículo en Inglés | LILACS | ID: biblio-977733

RESUMEN

Abstract Echinoderms have attracted the attention of scientists over the past few years after identifying a variety of unique structures endowed by interesting biological properties. However, the Moroccan coast biodiversity is still uninvestigated. In our ongoing attempts to valorize the rich Moroccan marine environment, this study aimed at assessing the antimicrobial activity of extracts obtained from three echinoderms Astropecten irregularis, Luidia sarsi and Ophiura albida against the human pathogens: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica and Bacillus subtilis. Moreover, their antioxidant activities were tested using standard methods in addition to the antidiabetic activity which has been evaluated in vitro against α-amylase and α-glucosidase enzymes. HPLC-DAD-QTOF-MS analysis revealed a significant content of some phenolic compounds such as pyrogallol, gallic, sinapic, ferulic, p-hydroxybenzoic and salicylic acids whose existence can be related to the endophytic fungi and/or dietary intake whereas GC-MS analysis exhibited diverse chemical structures such as cholesterol, oleic acid and glycerol 1-palmitate.

7.
Mater Sci Eng C Mater Biol Appl ; 71: 870-878, 2017 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-27987784

RESUMEN

Neuroblastoma is a pediatric neuroblastic tumor arising in the sympathetic nervous crest cells. A high grade of Neuroblastoma is characterized by a high urinary excretion of homovanillic acid and dopamine. In this work l-leucine modified Sol-Gel-Carbon electrode was used for a sensitive voltammetric determination of homovanillic acid and dopamine in urine. The electrochemical response characteristics were investigated by cyclic and differential pulse voltammetry; the modified electrode has shown an increase in the effective area of up to 40%, a well-separated oxidation peaks and an excellent electrocatalytic activity. High sensitivity and selectivity in the linear range of 0,4-100µML-1 of homovanillic acid and 10-120µML-1 of dopamine were also obtained. Moreover, a sub-micromolar limit of detection of 0.1µM for homovanillic acid and 1.0µM for the dopamine was achieved. Indeed, high reproducibility with simple preparation and regeneration of the electrode surface made this electrode very suitable for the determination of homovanillic acid and dopamine in pharmaceutical and clinical preparations. The mechanism of homovanillic acid and the electrochemical oxidation at l-leucine modified Sol-Gel-Carbon electrode is described out the B3P86/6-31+G(d,p) level of theory as implemented in Gaussian software.


Asunto(s)
Dopamina/metabolismo , Dopamina/orina , Ácido Homovanílico/orina , Leucina/química , Neuroblastoma/diagnóstico , Neuroblastoma/orina , Ácido Úrico/orina , Dopamina/análisis , Electrodos , Ácido Homovanílico/análisis , Humanos , Ácido Úrico/análisis
8.
Bioelectrochemistry ; 101: 84-91, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25179932

RESUMEN

New biosensors based on inhibition for the detection of cyanide and the comparison of the analytical performances of nine enzyme biosensor designs by using three different electrodes: Sonogel-Carbon, glassy carbon and gold electrodes were discussed. Three different horseradish peroxidase immobilization procedures with and without gold sononanoparticles were studied. The amperometric measurements were performed at an applied potential of -0.15V vs. Ag/AgCl in 50mM sodium acetate buffer solution pH=5.0. The apparent kinetic parameters (Kmapp, Vmaxapp) of immobilized HRP were calculated in the absence of inhibitor (cyanide) by using caffeic acid, hydroquinone, and catechol as substrates. The presence of gold sononanoparticles enhanced the electron transfer reaction and improved the analytical performance of the biosensors. The HRP kinetic interactions reveal non-competitive binding of cyanide with an apparent inhibition constant (Ki) of 2.7µM and I50 of 1.3µM. The determination of cyanide can be achieved in a dynamic range of 0.1-58.6µM with a detection limit of 0.03µM which is lower than those reported by previous studies. Hence this biosensing methodology can be used as a new promising approach for detecting cyanide.


Asunto(s)
Técnicas Biosensibles/métodos , Cianuros/análisis , Técnicas Electroquímicas/métodos , Enzimas Inmovilizadas/química , Peroxidasa de Rábano Silvestre/antagonistas & inhibidores , Nanopartículas/química , Técnicas Biosensibles/instrumentación , Ácidos Cafeicos/farmacología , Técnicas Electroquímicas/instrumentación , Electrodos , Enzimas Inmovilizadas/metabolismo , Oro , Peroxidasa de Rábano Silvestre/química , Peroxidasa de Rábano Silvestre/metabolismo , Concentración de Iones de Hidrógeno , Límite de Detección , Cianuro de Potasio/análisis , Cianuro de Potasio/farmacología
9.
J Hazard Mater ; 279: 348-55, 2014 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-25080156

RESUMEN

Amperometric hydrogen peroxide enzyme inhibition biosensors based on horseradish peroxidase (HRP) immobilised on electropolymerised neutral red (NR) or directly on the surface of carbon film electrodes (CFE) have been successfully applied to the determination of toxic Cr(III) and Cr(VI). Parameters influencing the performance of the biosensor including the enzyme immobilisation method, the amount of hydrogen peroxide, applied potential and electrolyte pH were optimised. The inhibition of horseradish peroxidase by the chromium species was studied under the optimised conditions. Results from the quantitative analysis of chromium ions are discussed in terms of detection limit, linear range and sensitivity. The HRP kinetic interactions reveal mixed binding of Cr(III) with I50=3.8µM and inhibition binding constant Ki=11.3µM at HRP/PNR/CFE biosensors and uncompetitive binding of Cr(VI) with I50=3.9µM and Ki=0.78µM at HRP/CFE biosensors in the presence of H2O2 substrate. Interferences from other heavy metal ions were studied and the inhibition show very good selectivity towards Cr(III) and Cr(VI).


Asunto(s)
Técnicas Biosensibles , Cromo/análisis , Peróxido de Hidrógeno/química , Rojo Neutro/química , Electroquímica , Electrodos , Enzimas Inmovilizadas/química , Peroxidasa de Rábano Silvestre/química , Concentración de Iones de Hidrógeno , Oxidación-Reducción
10.
Int J Food Microbiol ; 153(3): 314-22, 2012 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-22177229

RESUMEN

The technological and safety properties of 35 indigenous strains of Enterococcus faecium isolated from two Egyptian cheeses were characterised in order to determine their ability for rational manufacture of diversified and typical dairy products in the Middle East. A great diversity was observed within the 35 strains on the basis of their technological properties. A statistical analysis made it possible to distribute the 35 strains of E. faecium into different groups. Three groups were identified in terms of their acidification activity, measured by the Cinac system: a group of strains that quickly acidified milk, a second that moderately acidified milk, and a last cluster that revealed weak acidification activity. On the basis of texturing properties that were evaluated using a texturometer and a viscometer, a cluster of strains produced viscous and firm gels, a second cluster included fairly viscous, firm and cohesive gels, and a last group generated slightly viscous, but firm and very cohesive gels. By considering the aroma profiles that were determined by gas chromatography coupled with mass spectrometry, four clusters were identified. One cluster displayed a high dimethyl disulfide level, a second group of strains was highly aromatic, a third cluster led to typical "lactic" products, and the last cluster made it possible to obtain low aromatic products. None of the 35 strains proved to be ß-haemolytic on the basis of the characterisation of their safety properties. The resistance to 20 antibiotics was assessed by the disc diffusion method. The 35 isolates were sensitive to 12 antibiotics, and among them, one isolate was resistant to only two antibiotics (nalixidic acid and streptomycin). The resistance to eight antibiotics was strain-dependant. Finally, this study demonstrates that some indigenous strains of E. faecium displayed interesting technological properties for cheese manufacture, together with good safety characteristics. They could be useful for the manufacture of typical products in Egypt.


Asunto(s)
Queso/microbiología , Enterococcus faecium/aislamiento & purificación , Microbiología de Alimentos , Animales , Antibacterianos , Biodiversidad , Queso/análisis , Productos Lácteos , Farmacorresistencia Bacteriana , Egipto , Enterococcus/aislamiento & purificación , Enterococcus faecium/clasificación , Enterococcus faecium/fisiología , Medio Oriente , Leche/química , Estreptomicina , Viscosidad
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