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1.
J Mater Chem B ; 9(44): 9153-9161, 2021 11 17.
Artículo en Inglés | MEDLINE | ID: mdl-34694310

RESUMEN

Imidacloprid is the most widely used insecticide in agriculture and its intensive use over the last 30 years has caused a global concern due to its potentially toxic effects on the ecosystem. Considering the recent scientific interest in novel simple methods for imidacloprid analysis, we propose a label-free sensitive and specific localised surface plasmon resonance system for the detection of the insecticide based on 2D nanostructured metasurfaces with highly performing plasmonic properties. The specificity of the sensor proposed was achieved by covalent bio-functionalization of the metasurface using a smart and easy one-step procedure mediated by carbon disulphide. The biosensor produced was tested using a set of imidacloprid standard solutions showing a competitive limit of detection, lower than 1 ng mL-1. Our novel nanosensing configuration represents a valid and reliable solution to realize low-cost portable POC tests as an alternative to the laborious and expensive methods traditionally used for insecticide detection.


Asunto(s)
Insecticidas/análisis , Neonicotinoides/análisis , Nitrocompuestos/análisis , Resonancia por Plasmón de Superficie/métodos , Anticuerpos Inmovilizados/inmunología , Anticuerpos Monoclonales/inmunología , Oro/química , Insecticidas/inmunología , Límite de Detección , Nanopartículas del Metal/química , Neonicotinoides/inmunología , Nitrocompuestos/inmunología
2.
Food Chem ; 358: 129898, 2021 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-33933961

RESUMEN

The sensitive detection of pesticides in complex environment is important but still challenging in presence of organic-rich water sample and food matrix. Herein, we reported a nitrile-mediated SERS immunosensor for sensitive and optical anti-interference determination of imidacloprid. Raman tag contained CN bond could provide a sharp characteristic peak in the Raman-silent spectral window (1800 ~ 2800 cm-1), which could resist the optical noises from the fingerprint region (<1800 cm-1). Aucore-Agshell bimetallic nanocuboid (AuNR@Ag) connected with antigen and Raman tag was used as Raman probe, while Fe3O4 magnetic nanoparticle functionalized with anti-imidacloprid antibody was applied as signal enhancer. Owing to the specific recognition ability between antigen and antibody, the competitive system with imidacloprid was formed. Under the optimal condition, the linear relationship was developed in the range of 10-400 nM. Finally, the SERS immunosensor was successfully applied to determine imidacloprid in real samples with recoveries from 96.8% to 100.5%.


Asunto(s)
Análisis de los Alimentos/métodos , Inmunoensayo/métodos , Neonicotinoides/análisis , Nitrocompuestos/análisis , Espectrometría Raman/métodos , Contaminación de Alimentos/análisis , Oro/química , Inmunoensayo/instrumentación , Nanopartículas Magnéticas de Óxido de Hierro/química , Neonicotinoides/inmunología , Nitrocompuestos/inmunología , Plaguicidas/análisis , Plaguicidas/inmunología , Sensibilidad y Especificidad , Espectrometría Raman/instrumentación
3.
Talanta ; 209: 120465, 2020 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-31892037

RESUMEN

A direct competitive immunosensor for the electrochemical determination of Imidacloprid (IMD) pesticide on gold nanoparticle-modified screen-printed carbon electrodes (AuNP-SPCE) is here reported for the first time. Self-obtained specific monoclonal antibodies are immobilized on the AuNP-SPCE taking advantage of the AuNPs biofunctionalization abilities. In our biosensor design, free IMD in the sample competes with IMD conjugated with horseradish peroxidase (IMD-HRP) for the recognition by the antibodies. After that, 3,3',5,5'-Tetramethylbenzidine (TMB) is enzymatically oxidized by HRP, followed by the oxidized TMB reduction back at the surface of the SPCE. This process gives an associated catalytic current (analytical signal) that is inversely proportional to the IMD amount. The main parameters affecting the analytical signal have been optimized, reaching a good precision (repeatability with a RSD of 6%), accuracy (relative error of 6%), stability (up to one month), selectivity and an excellent limit of detection (LOD of 22 pmol L-1), below the maximum levels allowed by the legislation, with a wide response range (50-10000 pmol L-1). The detection through antibodies also allows to have an excellent selectivity against other pesticides potentially present in real samples. Low matrix effects were found when analysing IMD in tap water and watermelon samples. The electrochemical immunosensor was also validated with HPLC-MS/MS, the reference method used in official laboratories for IMD analysis, through statistical tests. Our findings make the electrochemical immunosensor as an outstanding method for the rapid and sensitive determination of IMD at the point-of-use.


Asunto(s)
Técnicas Biosensibles/métodos , Inmunoensayo/métodos , Nanopartículas del Metal/química , Neonicotinoides/análisis , Nitrocompuestos/análisis , Plaguicidas/análisis , Anticuerpos Inmovilizados/inmunología , Anticuerpos Monoclonales/inmunología , Armoracia/enzimología , Bencidinas/química , Citrullus/química , Agua Potable/análisis , Técnicas Electroquímicas/instrumentación , Técnicas Electroquímicas/métodos , Electrodos , Contaminación de Alimentos/análisis , Oro/química , Peroxidasa de Rábano Silvestre/química , Límite de Detección , Solanum lycopersicum/química , Neonicotinoides/inmunología , Nitrocompuestos/inmunología , Plaguicidas/inmunología , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/inmunología
4.
Allergy ; 75(2): 412-422, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31444907

RESUMEN

BACKGROUND: Food allergy is associated with a high personal health and economic burden. For immunomodulation toward tolerance, food compounds could be chemically modified, for example, by posttranslational protein nitration, which also occurs via diet-derived nitrating agents in the gastrointestinal tract. OBJECTIVE: We sought to analyze the effect of pretreatment with nitrated food proteins on the immune response in a mouse food allergy model and on human monocyte-derived dendritic cells (moDCs) and PBMCs. METHODS: The model allergen ovalbumin (OVA) was nitrated in different nitration degrees, and the secondary structures of proteins were determined by circular dichroism (CD). Allergy-preventive treatment with OVA, nitrated OVA (nOVA), and maximally nitrated OVA (nOVAmax) were performed before mice were immunized with or without gastric acid-suppression medication. Antibody levels, regulatory T-cell (Treg) numbers, and cytokine levels were evaluated. Human moDCs or PBMCs were incubated with proteins and evaluated for expression of surface markers, cytokine production, and proliferation of Th2 as well as Tregs. RESULTS: In contrast to OVA and nOVA, the conformation of nOVAmax was substantially changed. nOVAmax pretreated mice had decreased IgE as well as IgG1 and IgG2a levels and Treg numbers were significantly elevated, while cytokine levels remained at baseline level. nOVAmax induced a regulatory DC phenotype evidenced by a decrease of the activation marker CD86 and an increase in IL-10 production and was associated with a higher proliferation of memory Tregs. CONCLUSION: Oral pretreatment with highly nitrated proteins induces a tolerogenic immune response in the food allergy model and in human immune cells.


Asunto(s)
Alérgenos/química , Alérgenos/inmunología , Hipersensibilidad a los Alimentos/prevención & control , Inmunización/métodos , Nitrocompuestos/inmunología , Ovalbúmina/química , Ovalbúmina/inmunología , Linfocitos T Reguladores/inmunología , Administración Oral , Alérgenos/administración & dosificación , Animales , Donantes de Sangre , Células Cultivadas , Citocinas/metabolismo , Células Dendríticas/inmunología , Modelos Animales de Enfermedad , Femenino , Hipersensibilidad a los Alimentos/inmunología , Humanos , Tolerancia Inmunológica/inmunología , Inmunoglobulina E/metabolismo , Ratones , Ratones Endogámicos BALB C , Nitrocompuestos/administración & dosificación , Ovalbúmina/administración & dosificación , Transducción de Señal/inmunología
5.
Nutrients ; 11(10)2019 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-31618852

RESUMEN

Food proteins may get nitrated by various exogenous or endogenous mechanisms. As individuals might get recurrently exposed to nitrated proteins via daily diet, we aimed to investigate the effect of repeatedly ingested nitrated food proteins on the subsequent immune response in non-allergic and allergic mice using the milk allergen beta-lactoglobulin (BLG) as model food protein in a mouse model. Evaluating the presence of nitrated proteins in food, we could detect 3-nitrotyrosine (3-NT) in extracts of different foods and in stomach content extracts of non-allergic mice under physiological conditions. Chemically nitrated BLG (BLGn) exhibited enhanced susceptibility to degradation in simulated gastric fluid experiments compared to untreated BLG (BLGu). Gavage of BLGn to non-allergic animals increased interferon-γ and interleukin-10 release of stimulated spleen cells and led to the formation of BLG-specific serum IgA. Allergic mice receiving three oral gavages of BLGn had higher levels of mouse mast cell protease-1 (mMCP-1) compared to allergic mice receiving BLGu. Regardless of the preceding immune status, non-allergic or allergic, repeatedly ingested nitrated food proteins seem to considerably influence the subsequent immune response.


Asunto(s)
Alérgenos/inmunología , Lactoglobulinas/inmunología , Hipersensibilidad a la Leche/inmunología , Nitrocompuestos/inmunología , Animales , Línea Celular Tumoral , Quimasas/inmunología , Quimasas/metabolismo , Modelos Animales de Enfermedad , Femenino , Inmunoglobulina A/sangre , Inmunoglobulina A/inmunología , Interferón gamma/inmunología , Interferón gamma/metabolismo , Interleucina-10/inmunología , Interleucina-10/metabolismo , Ratones Endogámicos BALB C , Hipersensibilidad a la Leche/sangre , Estabilidad Proteica , Proteolisis , Ratas , Bazo/inmunología , Bazo/metabolismo , Tirosina/análogos & derivados , Tirosina/inmunología
6.
Analyst ; 144(9): 2936-2941, 2019 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-30920552

RESUMEN

Imidacloprid (IMD) is one of the most used pesticides worldwide as a systemic insecticide as well as for pest control and seed treatment. The toxic and potential carcinogenic character of IMD makes its monitoring of great relevance in the field of agriculture and environment, so sensitive methodologies for in field analysis are strongly required. In this context, we have developed a competitive immunoassay for the determination of IMD using specific monoclonal antibodies followed by electrochemical detection on screen-printed carbon electrodes (SPCE). The optimized immunosensor exhibited a good reproducibility (RSD of 9%) and a logarithmic response in the range 50-10 000 pM of IMD, with an estimated detection limit (LOD) of 24 pM, which was below the maximum levels allowed by the legislation. High-Performance Liquid Chromatography-Mass Spectrometry-Mass Spectrometry (HPLC-MSMS) and Enzyme-Linked Immunosorbent Assay (ELISA) analysis were also performed for comparison purposes, where the electrochemical immunosensor exhibited a wider range of response and a lower detection limit. Matrix effects below 6.5% were obtained using tap water samples. All these characteristics make our electrochemical immunosensor a valid and advantageous tool for the in field determination of IMD.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Técnicas Electroquímicas/métodos , Inmunoensayo/métodos , Insecticidas/análisis , Neonicotinoides/análisis , Nitrocompuestos/análisis , Animales , Armoracia/enzimología , Bencidinas/química , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Carbono/química , Bovinos , Compuestos Cromogénicos/química , Agua Potable/análisis , Técnicas Electroquímicas/instrumentación , Electrodos , Peroxidasa de Rábano Silvestre/química , Insecticidas/inmunología , Límite de Detección , Neonicotinoides/inmunología , Nitrocompuestos/inmunología , Oxidación-Reducción , Conejos , Reproducibilidad de los Resultados , Albúmina Sérica Bovina/química
7.
Free Radic Biol Med ; 129: 430-439, 2018 12.
Artículo en Inglés | MEDLINE | ID: mdl-30321702

RESUMEN

Alzheimer's disease (AD) is a progressive form of dementia characterized by increased production of amyloid-ß plaques and hyperphosphorylated tau protein, mitochondrial dysfunction, elevated oxidative stress, reduced protein clearance, among other. Several studies showed systemic modifications of immune and inflammatory systems due, in part, to decreased levels of CD3+ lymphocytes in peripheral blood in AD. Considering that oxidative stress, both in the brain and in the periphery, can influence the activation and differentiation of T-cells, we investigated the 3-nitrotyrosine (3-NT) proteome of blood T-cells derived from AD patients compared to non-demented (ND) subjects by using a proteomic approach. 3-NT is a formal protein oxidation and index of nitrosative stress. We identified ten proteins showing increasing levels of 3-NT in CD3+ T-cells from AD patients compared with ND subjects. These proteins are involved in energy metabolism, cytoskeletal structure, intracellular signaling, protein folding and turnover, and antioxidant response and provide new insights into the molecular mechanism that impact reduced T-cell differentiation in AD. Our results highlight the role of peripheral oxidative stress in T-cells related to immune-senescence during AD pathology focusing on the specific targets of protein nitration that conceivably can be suitable to further therapies. Further, our data demonstrate common targets of protein nitration between the brain and the periphery, supporting their significance as disease biomarkers.


Asunto(s)
Enfermedad de Alzheimer/diagnóstico , Linfocitos/química , Nitrocompuestos/inmunología , Proteoma/inmunología , Tirosina/análogos & derivados , Anciano , Anciano de 80 o más Años , Enfermedad de Alzheimer/inmunología , Enfermedad de Alzheimer/metabolismo , Antioxidantes/química , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Complejo CD3/genética , Complejo CD3/inmunología , Estudios de Casos y Controles , Separación Celular , Proteínas del Citoesqueleto/química , Proteínas del Citoesqueleto/inmunología , Metabolismo Energético/genética , Femenino , Expresión Génica , Humanos , Linfocitos/inmunología , Masculino , Persona de Mediana Edad , Nitrocompuestos/química , Nitrocompuestos/aislamiento & purificación , Estrés Nitrosativo , Estrés Oxidativo , Cultivo Primario de Células , Proteoma/genética , Proteoma/metabolismo , Transducción de Señal , Tirosina/metabolismo
8.
Anal Chem ; 87(5): 2952-8, 2015 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-25622025

RESUMEN

The multianalyte immunoassay (MIA) has attracted increasing attention due to its high sample throughput, short assay time, low sample consumption, and reduced overall cost. However, up to now, the reported MIA methods commonly require multiple antibodies since each antibody can recognize only one antigen. Herein, a novel bispecific monoclonal antibody (BsMcAb) that could bind methyl parathion and imidacloprid simultaneously was produced by a hybrid hybridomas strategy. A chemiluminescence (CL) reaction kinetics-resolved strategy was designed for MIA of methyl parathion and imidacloprid using the BsMcAb as the unique recognition reagent. Horseradish peroxidase (HRP) and alkaline phosphatase (ALP) were adopted as the signal probes to tag the haptens of the two pesticides due to their very different CL kinetic characteristics. After competitive immunoreactions, the HRP-tagged methyl parathion hapten and the ALP-tagged imidacloprid hapten were simultaneously bound to the BsMcAb since there were two different antigen-binding sites in it. Then, two CL reactions were simultaneously triggered by adding the CL coreactants, and the signals for methyl parathion and imidacloprid detections were collected at 0.6 and 1000 s, respectively. The linear ranges for methyl parathion and imidacloprid were both 1.0-500 ng/mL, with detection limits of 0.33 ng/mL (S/N = 3). The proposed method was successfully used to detect pesticides spiked in ginseng and American ginseng with acceptable recoveries of 80-118%. This proof-of-principle work demonstrated the feasibility of MIA using only one antibody.


Asunto(s)
Anticuerpos Biespecíficos/inmunología , Imidazoles/análisis , Inmunoensayo/métodos , Mediciones Luminiscentes/métodos , Metil Paratión/análisis , Nitrocompuestos/análisis , Panax/química , Plaguicidas/análisis , Fosfatasa Alcalina/metabolismo , Animales , Anticuerpos Biespecíficos/química , Anticuerpos Biespecíficos/metabolismo , Femenino , Peroxidasa de Rábano Silvestre/metabolismo , Hibridomas , Imidazoles/inmunología , Imidazoles/metabolismo , Inmunización , Insecticidas/análisis , Insecticidas/inmunología , Insecticidas/metabolismo , Límite de Detección , Metil Paratión/inmunología , Metil Paratión/metabolismo , Ratones , Ratones Endogámicos BALB C , Neonicotinoides , Nitrocompuestos/inmunología , Nitrocompuestos/metabolismo
9.
Int Immunopharmacol ; 23(1): 283-7, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-25241247

RESUMEN

Nitrated fatty acids (NFAs), thought to be produced by nonenzymatic reactions of endogenous nitric oxide (NO) with naturally present unsaturated fatty acids, have recently been identified as one of the largest single pools of biologically active NO derivatives in human plasma. As the biological role of NFAs is unknown, initial in vitro studies have shown them to be potent suppressors of inflammatory responses. The aim of the study was to collect all the literature on NFAs and its interactions with peroxisome proliferator-activated receptor gamma (PPAR-γ) and review in detail the anti-inflammatory properties of PPAR-γ interceded by NFAs. A literature survey was performed using PubMed and ScienceDirect to gather complete information on NFAs and their interactions with PPAR-γ. An exhaustive literature survey revealed that NFAs found in human plasma and urine comprises a class of cell signaling mediators that can activate PPAR-γ within its physiological concentration. NFAs exhibit anti-inflammatory and anti-fibrotic effects through PPAR-γ activation in various in vitro models tested. Besides its role in inflammation other properties of NFAs such as inhibition of enzymes, inducer of gene expression, etc., were discussed. NFAs are good electrophiles with pleiotropic biological activities. Hence NFAs can be treated as potent drug candidates.


Asunto(s)
Ácidos Grasos/inmunología , Inflamación/inmunología , Nitrocompuestos/inmunología , PPAR gamma/inmunología , Animales , Antiinflamatorios/uso terapéutico , Descubrimiento de Drogas , Ácidos Grasos/química , Ácidos Grasos/farmacología , Humanos , Inmunomodulación , Inflamación/tratamiento farmacológico , Óxido Nítrico/metabolismo , Nitrocompuestos/química , Nitrocompuestos/farmacología , Transducción de Señal/inmunología
10.
Sensors (Basel) ; 12(11): 15858-72, 2012 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-23202236

RESUMEN

Enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody (MoAb) was developed for the neonicotinoid insecticide clothianidin. A new clothianidin hapten (3-[5-(3-methyl-2-nitroguanidinomethyl)-1,3-thiazol-2-ylthio] propionic acid) was synthesized and conjugated to keyhole limpet hemocyanin, and was used for monoclonal antibody preparation. The resulting MoAb CTN-16A3-13 was characterized by a direct competitive ELISA (dc-ELISA). The 50% of inhibition concentration value with clothianidin was 4.4 ng/mL, and the working range was 1.5–15 ng/mL. The antibody showed high cross-reactivity (64%) to dinotefuran among the structurally related neonicotinoid insecticides. The recovery examinations of clothianidin for cucumber, tomato and apple showed highly agreement with the spiked concentrations; the recovery rate was between 104% and 124% and the coefficient of variation value was between 1.8% and 15%. Although the recovery rate of the dc-ELISA was slightly higher than that of HPLC analysis, the difference was small enough to accept the dc-ELISA as a useful method for residue analysis of clothianidin in garden crops.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Guanidinas/análisis , Insecticidas/análisis , Nitrocompuestos/análisis , Tiazoles/análisis , Cromatografía Líquida de Alta Presión , Guanidinas/inmunología , Insecticidas/inmunología , Neonicotinoides , Nitrocompuestos/inmunología , Tiazoles/inmunología
11.
J Sci Food Agric ; 92(6): 1253-60, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22083888

RESUMEN

BACKGROUND: Imidacloprid is one of the main neonicotinoid insecticides widely used in agriculture owing its broad spectrum of activity and low bioaccumulation. However, imidacloprid is toxic to honey bees and other beneficial organisms, and its residues may occur in environmental and food samples, posing a potential hazard to consumers. In this study the imidacloprid derivative bearing a three-atom length spacer was synthesized and coupled to carrier proteins. Highly sensitive and specific polyclonal antibodies against imidacloprid were successfully produced and the polyclonal antibody-based enzyme-linked immunosorbent assay (pAb-ELISA) was developed. RESULTS: The ELISA standard curve was constructed within the concentration range 0.1-100 ng mL(-1). The IC(50) value for nine standard curves was in the range 1.2-3.0 ng mL(-1) and the limit of detection was 0.03-0.16 ng mL(-1). The sensitivity of the assay was one order of magnitude higher than that in most published papers. There was almost no cross-reactivity of the antibody with four structurally related compounds (acetamiprid, nicotine, clothianidin and nitenpyram) and six other compounds, indicating that the assay displays not only high sensitivity but also high specificity. No detectable imidacloprid was found in 11 collected environmental and food samples by the assay. For imidacloprid-spiked samples, acceptable recoveries of 73.4-94.4% and intra-assay coefficients of variation of 2.2-12.8% were obtained. The assay was also validated with high-performance liquid chromatography (HPLC) and a good correlation of ELISA with HPLC was achieved. CONCLUSION: The proposed ELISA provides a sensitive, specific, simple and cost-effective quantitative/screening method for detecting imidacloprid in environmental and food samples.


Asunto(s)
Anticuerpos , Monitoreo del Ambiente/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Contaminación de Alimentos/análisis , Imidazoles/análisis , Insecticidas/análisis , Nitrocompuestos/análisis , Animales , Abejas , Cromatografía Líquida de Alta Presión , Conservación de los Recursos Naturales , Reacciones Cruzadas , Humanos , Imidazoles/inmunología , Concentración 50 Inhibidora , Insecticidas/inmunología , Límite de Detección , Neonicotinoides , Nitrocompuestos/inmunología
12.
Sheng Wu Gong Cheng Xue Bao ; 27(6): 943-51, 2011 Jun.
Artículo en Chino | MEDLINE | ID: mdl-22034824

RESUMEN

To produce high-affinity monoclonal antibodies against pesticide imidacloprid, we synthesized the haptens 1-[(6-Carboxylethylthio-3-pyridinyl) methyl] -N-nitro-imidazolidinimine (named as H1) and 1-[(6-Chloro-3-pyridinyl) methyl]-3-carboxylpropyl-N-nitro-2-imidazolidinimine (termed as H2). And then the haptens were coupled to bovine serum albumin (BSA) and ovalbumin (OVA) for immunogen (H1-BSA) and coating antigen (H2-OVA) respectively by NHS ester method. BALB/c mice were immunized with H1-BSA conjugate. We obtained two hybridoma cell lines 2F11/A9 and 2G6/G12 secreting antibody specific for imidacloprid from the conventional hybridoma technology. The result showed that the subtypes of obtained monoclonal antibodies were IgG3 and IgG1, respectively, and the titers of ascites were up to 1:128 000. The indirect competitive ELISA indicated the IC50 values of 5.3 and 28.3 ng/mL with detection limits of 1.1 ng/mL and 7.7 ng/mL, respectively. Two monoclonal antibodies had no apparent cross reactivity with six analogous compounds. Thus, two prepared monoclonal antibodies had a very high affinity and specificity, and it could be used to develop ELISA for rapid determination of imidacloprid residue and laid a solid foundation for research and development of products for immunoassay.


Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Imidazoles/inmunología , Insecticidas/inmunología , Nitrocompuestos/inmunología , Animales , Especificidad de Anticuerpos , Hibridomas/metabolismo , Ratones , Ratones Endogámicos BALB C , Neonicotinoides
13.
Talanta ; 84(4): 1107-11, 2011 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-21530785

RESUMEN

The analytical performance of a kit-based enzyme-linked immunosorbent assay (ELISA) for the determination of a neonicotinoid insecticide dinotefuran residue in rice samples is addressed. The sensitivity (I(50) value) was 5.4 ng/mL, with the limit of detection, 0.6 ng/mL and the dynamic range from 1.0 to 30 ng/mL. The ELISA showed substantially high specificity toward dinotefuran besides clothianidin (184%). For rice samples, dinotefuran was extracted with methanol and the extracts were directly determined with the ELISA because of no significant matrix interference. Good recoveries were observed and ranged from 92.5% to 113.2% with coefficients of variation below 10%. The results obtained with the ELISA correlated well with those by the HPLC method for rice samples (r>0.98). These findings strongly indicate that the evaluated and validated ELISA has a potential utility in a quick, simple, and reliable residue analysis, especially a screening method before shipment contributing to food safety.


Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Contaminación de Alimentos/análisis , Guanidinas/análisis , Nitrocompuestos/análisis , Oryza/química , Residuos de Plaguicidas/análisis , Cromatografía Líquida de Alta Presión , Reacciones Cruzadas , Guanidinas/inmunología , Neonicotinoides , Nitrocompuestos/inmunología , Residuos de Plaguicidas/inmunología , Reproducibilidad de los Resultados , Solventes/química , Factores de Tiempo
14.
Bioconjug Chem ; 22(4): 633-41, 2011 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-21446744

RESUMEN

Regulation of enzyme activity either by its substrates or by effectors is generally known as allostery. However, it has been considered hard to alter its effector specificity, despite its potential utility as a sensitive molecular sensor. To this end, we made fusion proteins consisting of an antibody variable region Fv and a circularly permutated TEM-1 ß-lactamase cpBLA. Two expression vectors encoding Fv-cpBLA with different antigen specificities were made, in which cpBLA was inserted into the linker region of the single chain Fv that specifically binds either bone-related disease marker osteocalcin (BGP) C-terminal peptide or neonicotinoid insecticide imidacloprid (ICP). The cpBLA having new termini near the active site was activated upon binding with its cognate antigen, owing to the stabilization of tethered Fv by bound antigen. As a result, both Fv-cpBLA showed specific antigen binding as well as antigen-induced enhancement in catalytic activity. Moreover, E. coli cells expressing Fv-cpBLA for ICP showed ICP concentration dependent growth in the medium containing ampicillin. The system was also applied to select for Fv-cpBLA linker mutants that confer faster growth. This will be the first of an antibody-based small molecule indicator enzyme.


Asunto(s)
Imidazoles/inmunología , Fragmentos de Inmunoglobulinas/inmunología , Nitrocompuestos/inmunología , beta-Lactamasas/inmunología , Imidazoles/química , Fragmentos de Inmunoglobulinas/química , Neonicotinoides , Nitrocompuestos/química , beta-Lactamasas/química
15.
Proc Natl Acad Sci U S A ; 107(41): 17527-32, 2010 Oct 12.
Artículo en Inglés | MEDLINE | ID: mdl-20876120

RESUMEN

Neonicotinoid insecticides control crop pests based on their action as agonists at the insect nicotinic acetylcholine receptor, which accepts chloropyridinyl- and chlorothiazolyl-analogs almost equally well. In some cases, these compounds have also been reported to enhance plant vigor and (a)biotic stress tolerance, independent of their insecticidal function. However, this mode of action has not been defined. Using Arabidopsis thaliana, we show that the neonicotinoid compounds, imidacloprid (IMI) and clothianidin (CLO), via their 6-chloropyridinyl-3-carboxylic acid and 2-chlorothiazolyl-5-carboxylic acid metabolites, respectively, induce salicylic acid (SA)-associated plant responses. SA is a phytohormone best known for its role in plant defense against pathogens and as an inducer of systemic acquired resistance; however, it can also modulate abiotic stress responses. These neonicotinoids effect a similar global transcriptional response to that of SA, including genes involved in (a)biotic stress response. Furthermore, similar to SA, IMI and CLO induce systemic acquired resistance, resulting in reduced growth of a powdery mildew pathogen. The action of CLO induces the endogenous synthesis of SA via the SA biosynthetic enzyme ICS1, with ICS1 required for CLO-induced accumulation of SA, expression of the SA marker PR1, and fully enhanced resistance to powdery mildew. In contrast, the action of IMI does not induce endogenous synthesis of SA. Instead, IMI is further bioactivated to 6-chloro-2-hydroxypyridinyl-3-carboxylic acid, which is shown here to be a potent inducer of PR1 and inhibitor of SA-sensitive enzymes. Thus, via different mechanisms, these chloropyridinyl- and chlorothiazolyl-neonicotinoids induce SA responses associated with enhanced stress tolerance.


Asunto(s)
Arabidopsis/inmunología , Ascomicetos/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Guanidinas/farmacología , Imidazoles/farmacología , Insecticidas/farmacología , Nitrocompuestos/farmacología , Ácido Salicílico/inmunología , Tiazoles/farmacología , Arabidopsis/microbiología , Cromatografía Líquida de Alta Presión , Regulación de la Expresión Génica de las Plantas/inmunología , Guanidinas/química , Guanidinas/inmunología , Imidazoles/química , Imidazoles/inmunología , Insecticidas/química , Insecticidas/inmunología , Espectrometría de Masas , Estructura Molecular , Neonicotinoides , Nitrocompuestos/química , Nitrocompuestos/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ácido Salicílico/metabolismo , Tiazoles/química , Tiazoles/inmunología
16.
Free Radic Biol Med ; 46(8): 989-1003, 2009 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-19200454

RESUMEN

The signaling mediators nitric oxide ( NO) and oxidized lipids, once viewed to transduce metabolic and inflammatory information via discrete and independent pathways, are now appreciated as interdependent regulators of immune response and metabolic homeostasis. The interactions between these two classes of mediators result in reciprocal control of mediator synthesis that is strongly influenced by the local chemical environment. The relationship between the two pathways extends beyond coregulation of NO and eicosanoid formation to converge via the nitration of unsaturated fatty acids to yield nitro derivatives (NO(2)-FA). These pluripotent signaling molecules are generated in vivo as an adaptive response to oxidative inflammatory conditions and manifest predominantly anti-inflammatory signaling reactions. These actions of NO(2)-FA are diverse, with these species serving as a potential chemical reserve of NO, reacting with cellular nucleophiles to posttranslationally modify protein structure, function, and localization. In this regard these species act as potent endogenous ligands for peroxisome proliferator-activated receptor gamma. Functional consequences of these signaling mechanisms have been shown in multiple model systems, including the inhibition of platelet and neutrophil functions, induction of heme oxygenase-1, inhibition of LPS-induced cytokine release in monocytes, increased insulin sensitivity and glucose uptake in adipocytes, and relaxation of preconstricted rat aortic segments. These observations have propelled further in vitro and in vivo studies of mechanisms of NO(2)-FA signaling and metabolism, highlighting the therapeutic potential of this class of molecules as anti-inflammatory drug candidates.


Asunto(s)
Ácidos Grasos/metabolismo , Inflamación , Óxido Nítrico/metabolismo , Nitrocompuestos/metabolismo , Transducción de Señal/inmunología , Animales , Citocinas/metabolismo , Ácidos Grasos/inmunología , Regulación de la Expresión Génica , Hemo-Oxigenasa 1/genética , Hemo-Oxigenasa 1/inmunología , Hemo-Oxigenasa 1/metabolismo , Humanos , Óxido Nítrico/inmunología , Nitrocompuestos/inmunología , Estrés Oxidativo , PPAR gamma/inmunología , PPAR gamma/metabolismo , Procesamiento Proteico-Postraduccional/inmunología , Ratas , Vasodilatación
17.
Immunopharmacol Immunotoxicol ; 28(3): 491-500, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16997797

RESUMEN

A nasal formulation of mometasone furoate (MF) is advantageous in avoiding systemic activity characteristic of glucocorticoids when it is applied topically. To confirm antiallergic effects of this glucocorticoid formulation elaborately, we investigated whether the drug can suppress the production of IgE antibodies and related cytokines. It we showed that IgE production induced in mice immunized via intranasal route was significantly reduced when the mice were administered MF intranasally. Further, MF was effective in inhibiting production of type-2 helper T cell cytokines in vivo and in vitro. These results provide a immunopharmacological basis for clinical efficacy of this drug.


Asunto(s)
Formación de Anticuerpos/efectos de los fármacos , Inmunoglobulina E/inmunología , Interleucina-4/biosíntesis , Pregnadienodioles/farmacología , Administración Intranasal , Animales , Antialérgicos/administración & dosificación , Antialérgicos/inmunología , Antialérgicos/farmacología , Formación de Anticuerpos/inmunología , Línea Celular , Hipersensibilidad a las Drogas/inmunología , Hipersensibilidad a las Drogas/metabolismo , Hipersensibilidad a las Drogas/prevención & control , Inmunización/métodos , Inmunoglobulina E/metabolismo , Inmunoglobulina G/inmunología , Inmunoglobulina G/metabolismo , Inyecciones Subcutáneas , Masculino , Ratones , Ratones Endogámicos BALB C , Furoato de Mometasona , Nitrocompuestos/administración & dosificación , Nitrocompuestos/inmunología , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Pregnadienodioles/administración & dosificación , Pregnadienodioles/inmunología , Células Th2/efectos de los fármacos , Células Th2/inmunología , Células Th2/metabolismo
18.
J Immunol Methods ; 310(1-2): 159-70, 2006 Mar 20.
Artículo en Inglés | MEDLINE | ID: mdl-16499922

RESUMEN

The emission of soot during combustion processes used in transportation, manufacturing, and power generation is of increasing concern because of its serious adverse health effects. In particular, the ability to modulate the immune system has recently been established. In the present investigation, an artificial soot sample that was prepared by fragmentation of acetylene in a laser-induced plasma was used as an antigen for the immunization of a rabbit. A highly sensitive competition curve in an indirect competitive ELISA using a benzo[a]pyrene-BSA conjugate as a coating antigen could be constructed for benzo[a]pyrene with an IC50 of 2.94 mug/l (11.65 nmol/l). In contrast to the high affinity, the soot antiserum dilution (antibody titer) of 1:750 was rather low. The cross-reactivity was tested with 16 parent polycyclic aromatic hydrocarbons, 7 nitrated polycyclic aromatics, and 3 methylated, hydroxylated or butyric acid derivatives. The results obtained suggest that the vertebrate immune system can respond to an immunization with soot by the generation of high affinity IgG class antibodies against polycyclic aromatics. It is likely that antibodies are raised against the molecular structures which form the framework of the soot particles and not against adsorbed and extractable polyaromatic compounds. The experiments suggest that if soot is considered a T-independent antigen, the isotype switch, essentially from IgM to IgG, could have been caused by co-inoculation with a T-dependent antigen, i.e., mycobacteria contained in Freunds complete adjuvant. However, at the cellular level the mechanism remains to be uncovered.


Asunto(s)
Carbono/inmunología , Inmunoglobulina G/inmunología , Inmunoglobulina M/inmunología , Nitrocompuestos/inmunología , Hidrocarburos Policíclicos Aromáticos/inmunología , Acetileno/química , Animales , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/sangre , Inmunoglobulina M/biosíntesis , Inmunoglobulina M/sangre , Concentración 50 Inhibidora , Masculino , Microscopía Electrónica de Transmisión , Conejos
19.
J Agric Food Chem ; 51(7): 1823-30, 2003 Mar 26.
Artículo en Inglés | MEDLINE | ID: mdl-12643637

RESUMEN

An enzyme-linked immunosorbent assay (ELISA) was developed for the neonicotinoid insecticide thiamethoxam, 3-(2-chlorothiazol-5-ylmethyl)-5-methyl-4-nitroimino-1,3,5-oxadiazinane. Three antisera were raised from rabbits immunized with the hapten-KLH conjugate. On the basis of the computational analysis of hapten candidates, the hapten with a spacer arm on the thiazolyl ring of thiamethoxam was synthesized to elicit thiamethoxam-specific antisera. The hapten was 3-[2-(2-carboxyethylthio)-5-ylmethyl]-5-methyl-4-nitroimino-1,3,5-oxadiazinane. Antisera were characterized with indirect competitive ELISA. Cross-reactivity and effects of organic solvents, pH, and ionic strengths were evaluated. The antiserum was specific for thiamethoxam and tolerant of up to 5% acetonitrile and 5% acetone. Various ionic strengths and pH values in the tested ranges had negligible effect on the assay performance. Under the optimized conditions, the half-maximal inhibition concentration (IC(50)) and the limit of detection were approximately 9.0 and 0.1 microg/L of thiamethoxam, respectively. ELISA analysis of stream and tap water samples showed an excellent correlation with the fortification levels.


Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Insecticidas/análisis , Nitrocompuestos/análisis , Oxazinas/análisis , Animales , Especificidad de Anticuerpos , Haptenos/química , Haptenos/inmunología , Hemocianinas/inmunología , Sueros Inmunes/inmunología , Modelos Moleculares , Estructura Molecular , Neonicotinoides , Nitrocompuestos/química , Nitrocompuestos/inmunología , Oxazinas/química , Oxazinas/inmunología , Conejos , Tiametoxam , Tiazoles , Contaminantes del Agua/análisis
20.
Zh Mikrobiol Epidemiol Immunobiol ; (5): 15-20, 1984 May.
Artículo en Ruso | MEDLINE | ID: mdl-6540021

RESUMEN

The immunogenic properties of the dihapten and monohapten derivatives of polyethylene glycol with different nitroaromatic groupings were studied. 2,4-dinitrophenyl, 2, 4, 6-trinitrophenyl and trinitrophenyl-ethyl groupings were used as hapten groups. The injection of monohapten compounds was found to induce the accumulation of antibody-forming cells secreting antibodies to trinitrophenyl in the spleen of normal and athymic nude mice. As early as on day 3 the number of antibody-forming cells considerably exceeded their background level, the process of B-cell activation being, to a certain extent, thymus-independent. Dihapten compounds were not immunologically active. The effect rendered by the nitroaromatic derivatives of polyethylene glycol, revealed in this study, is linked with the known capacity of polyethylene glycol to adsorb on the surface of cell membranes.


Asunto(s)
Antígenos/inmunología , Haptenos/inmunología , Nitrocompuestos/inmunología , Nitrobencenos/inmunología , Polietilenglicoles/inmunología , Animales , Células Productoras de Anticuerpos/efectos de los fármacos , Antígenos T-Independientes/inmunología , Dinitrobencenos/inmunología , Relación Dosis-Respuesta Inmunológica , Eritrocitos/efectos de los fármacos , Eritrocitos/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Desnudos , Peso Molecular , Trinitrobencenos/inmunología
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