RESUMEN
Zearalenone (ZEN) is an extremely hazardous chemical widely existing in cereals, and its high-sensitivity detection possesses significant significance to human health. Here, the cathodic aggregation-induced electrochemiluminescence (AIECL) performance of tetraphenylethylene nanoaggregates (TPE NAs) was modulated by solvent regulation, based on which an electrochemiluminescence (ECL) aptasensor was constructed for sensitive detection of ZEN. The aggregation state and AIECL of TPE NAs were directly and simply controlled by adjusting the type of organic solvent and the fraction of water, which solved the current shortcomings of low strength and weak stability of the cathode ECL signal for TPE. Impressively, in a tetrahydrofuran-water mixed solution (volume ratio, 6:4), the relative ECL efficiency of TPE NAs reached 16.03%, which was 9.2 times that in pure water conditions, and the maximum ECL spectral wavelength was obviously red-shifted to 617 nm. In addition, "H"-shape DNA structure-mediated dual-catalyzed hairpin self-assembly (H-D-CHA) with higher efficiency by the synergistic effect between the two CHA reactions was utilized to construct a sensitive ECL aptasensor for ZEN analysis with a low detection limit of 0.362 fg/mL. In conclusion, solvent regulation was a simple and efficient method for improving the performance of AIECL materials, and the proposed ECL aptasensor had great potential for ZEN monitoring in food safety.
Asunto(s)
Técnicas Electroquímicas , Electrodos , Mediciones Luminiscentes , Solventes , Zearalenona , Zearalenona/análisis , Zearalenona/química , Solventes/química , Estilbenos/química , Límite de Detección , Técnicas Biosensibles , Aptámeros de Nucleótidos/químicaRESUMEN
Zearalenone (ZEN) and its derivatives are estrogenic mycotoxins known to pose significant health threats to humans and animals. Especially, the derivative α-zearalanol (α-ZAL) is over 10 times more toxic than ZEN. Simultaneous degradation of ZEN and its derivatives, especially α-ZAL, using ZEN lactone hydrolases (ZHDs) is a promising solution to eliminate their potential hazards to food safety. However, most available ZHDs exhibit limited activity toward the more toxic α-ZAL compared to ZEN. Here, we identified a broad-substrate spectrum ZHD, named ZHDAY3, from Exophiala aquamarina CBS 119918, which could not only efficiently degrade ZEN but also exhibited 73% relative activity toward α-ZAL. Through rational design, we obtained the ZHDAY3(N153H) mutant, which exhibited the highest specific activity (253.3 ± 4.3 U/mg) reported so far for degrading α-ZAL. Molecular docking, structural comparative analysis, and kinetic analysis collectively suggested that the shorter distance between the side chain of the catalytic residue His242 and the lactone bond of α-ZAL and the increased binding affinity to the substrate were mainly responsible for the improved catalytic activity of ZHDAY3(N153H) mutant. This mechanism was further validated through additional molecular docking of 18 mutants and experimental verification of six mutants.IMPORTANCEThe mycotoxins zearalenone (ZEN) and its derivatives pose a significant threat to food safety. Here, we present a highly promising ZEN lactone hydrolase (ZHD), ZHDAY3, which is capable of efficiently degrading both ZEN and the more toxic derivative α-ZAL. Next, the ZHDAY3(N153H) mutant obtained by single-point mutation exhibited the highest specific activity for degrading α-ZAL reported thus far. We further elucidated the molecular mechanisms underlying the enhanced hydrolytic activity of ZHDAY3(N153H) toward α-ZAL. These findings represent the first investigation on the molecular mechanism of ZHDs against α-ZAL and are expected to provide a significant reference for further rational engineering of ZHDs, which will ultimately contribute to addressing the health risks and food safety issues posed by ZEN-like mycotoxins.
Asunto(s)
Micotoxinas , Zearalenona , Zeranol , Humanos , Animales , Zearalenona/química , Zearalenona/metabolismo , Zeranol/química , Zeranol/metabolismo , Lactonas , Mutación Puntual , Hidrolasas/metabolismo , Simulación del Acoplamiento Molecular , Cinética , Micotoxinas/metabolismoRESUMEN
BACKGROUND: The mycotoxin zearalenone (ZEA) produced by toxigenic fungi is widely present in cereals and its downstream products. The danger of ZEA linked to various human health issues has attracted increasing attention. Thus, powerful ZEA-degrading or detoxifying strategies are urgently needed. Biology-based detoxification methods are specific, efficient, and environmentally friendly and do not lead to negative effects during cereal decontamination. Among these, ZEA detoxification using degrading enzymes was documented to be a promising strategy in broad research. Here, two efficient ZEA-degrading lactonases from the genus Gliocladium, ZHDR52 and ZHDP83, were identified for the first time. This work studied the degradation capacity and properties of ZEA using purified recombinant ZHDR52 and ZHDP83. RESULTS: According to the ZEA degradation study, transformed Escherichia coli BL21(DE3) PLySs cells harboring the zhdr52 or zhdp83 gene could transform 20 µg/mL ZEA within 2 h and degrade > 90% of ZEA toxic derivatives, α/ß-zearalanol and α/ß-zearalenol, within 6 h. Biochemical analysis demonstrated that the optimal pH was 9.0 for ZHDR52 and ZHDP83, and the optimum temperature was 45 °C. The purified recombinant ZHDR52 and ZHDP83 retained > 90% activity over a wide range of pH values and temperatures (pH 7.0-10.0 and 35-50 °C). In addition, the specific activities of purified ZHDR52 and ZHDP83 against ZEA were 196.11 and 229.64 U/mg, respectively. The results of these two novel lactonases suggested that, compared with ZHD101, these two novel lactonases transformed ZEA into different products. The slight position variations in E126 and H242 in ZDHR52/ZEA and ZHDP83/ZEA obtained via structural modelling may explain the difference in degradation products. Moreover, the MCF-7 cell proliferation assay indicated that the products of ZEA degradation using ZHDR52 and ZHDP83 did not exhibit estrogenic activity. CONCLUSIONS: ZHDR52 and ZHDP83 are alkali ZEA-degrading enzymes that can efficiently and irreversibly degrade ZEA into non-estrogenic products, indicating that they are potential candidates for commercial application. This study identified two excellent lactonases for industrial ZEA detoxification.
Asunto(s)
Gliocladium , Zearalenona , Zeranol/análogos & derivados , Humanos , Zearalenona/química , Gliocladium/metabolismo , BiotransformaciónRESUMEN
In view of the animal feeds inevitably contaminated by multiple mycotoxins, eco-friendly and efficient palygorskite-montmorillonite (Pal-Mt) materials were prepared to remove polar aflatoxin B1 (AFB1) and weak polar zearalenone (ZEN) from mixed mycotoxins aqueous solution. The adsorption properties and bonding mechanisms between Pal-Mt materials and mycotoxins (AFB1 and ZEN) were investigated systematically. The as-prepared Pal-Mt showed excellent adsorption capacity for AFB1 and ZEN in single- and binary-mycotoxin systems, indicating the effectiveness of Pal-Mt acting as multiple mycotoxin adsorbents. The kinetics of adsorption for ZEN was fast due to the adsorption on the external surface (film and intraparticle diffusion), while AFB1 molecules permeated into mesopores after the external adsorption for the more planar structure. Adsorption isotherms demonstrated that heterogeneous surface adsorption appeared between Pal-Mt and AFB1, and monolayer adsorption occurred on Pal-Mt and ZEN for different polarities of mycotoxins. Thermodynamic parameters illustrated that the adsorption process of both AFB1 and ZEN onto Pal-Mt was spontaneous and endothermic. The adsorption mechanism studies suggested that hydrogen bonding, electrostatic attraction, calcium bridging linkage, and ion-dipole played fundamental roles in the interaction between Pal-Mt and these two mycotoxins.
Asunto(s)
Micotoxinas , Zearalenona , Animales , Zearalenona/química , Aflatoxina B1/química , Bentonita/química , Micotoxinas/química , AdsorciónRESUMEN
Zearalenone (ZEN) is one of the most prevalent estrogenic mycotoxins, is produced mainly by the Fusarium family of fungi, and poses a risk to the health of animals. Zearalenone hydrolase (ZHD) is an important enzyme capable of degrading ZEN into a non-toxic compound. Although previous research has investigated the catalytic mechanism of ZHD, information on its dynamic interaction with ZEN remains unknown. This study aimed to develop a pipeline for identifying the allosteric pathway of ZHD. Using an identity analysis, we identified hub genes whose sequences can generalize a set of sequences in a protein family. We then utilized a neural relational inference (NRI) model to identify the allosteric pathway of the protein throughout the entire molecular dynamics simulation. The production run lasted 1 microsecond, and we analyzed residues 139-222 for the allosteric pathway using the NRI model. We found that the cap domain of the protein opened up during catalysis, resembling a hemostatic tape. We used umbrella sampling to simulate the dynamic docking phase of the ligand-protein complex and found that the protein took on a square sandwich shape. Our energy analysis, using both molecular mechanics/Poisson-Boltzmann (Generalized-Born) surface area (MMPBSA) and Potential Mean Force (PMF) analysis, showed discrepancies, with scores of -8.45 kcal/mol and -1.95 kcal/mol, respectively. MMPBSA, however, obtained a similar score to that of a previous report.
Asunto(s)
Micotoxinas , Zearalenona , Zearalenona/química , Hidrolasas/química , Simulación de Dinámica Molecular , Micotoxinas/metabolismo , Movimiento (Física)RESUMEN
Raw Ca-based montmorillonite (MMT) was treated by H2SO4, calcination and organic compounds (hexadecyltrimethyl ammonium bromide (HTAB), cetylpyridinium chloride (CPC) and chitosan (CTS)), respectively. The modified montmorillonites were characterized by different methods and their adsorption performances for three mycotoxins (Aflatoxin B1 (AFB1), zearalenone (ZEA) and deoxynivalenol (DON)) were evaluated at pH = 2.8 and 8.0, respectively. The results indicate that surfactants (CPC and HTAB) intercalation is the most efficient modification, which obviously improves the adsorption performance of montmorillonite for mycotoxins, with adsorption efficiency of above 90% for AFB1 and ZEA whether under acid or alkaline conditions, due to the increase in basal spacing and the improvement of hydrophobicity. Moreover, the adsorption efficiencies of AFB1 and ZEA over CPC-modified montmorillonite (CPC-AMMT-3) coexisting with vitamin B6 or lysine are still at a high level (all above 94%). All modified montmorillonites, however, have low adsorption efficiency for DON, with somewhat spherical molecular geometry.
Asunto(s)
Aflatoxina B1/química , Bentonita/química , Inactivación Metabólica , Tricotecenos/química , Zearalenona/química , Ácidos/química , Adsorción , Calcio/química , Micotoxinas/química , TemperaturaRESUMEN
Zearalenone (ZEN), one of the most dangerous mycotoxins, causes enormous economic losses in the food and feed industries. To solve the problem of ZEN pollution, ZEN detoxifying enzymes are in emergent need. In this study, a zearalenone lactonohydrolase from Trichoderma aggressivum, denoted as ZHD-P, was heterologously expressed and characterized. The intracellular ZHD-P from E. coli BL21(DE3) exhibited high activity for ZEN degradation (191.94 U/mg), with the optimal temperature and pH of 45 °C and 7.5-9.0, respectively. With excellent temperature stability, the intracellular ZHD-P retained 100% activity when it was incubated at 25-40 °C for 1 h. Furthermore, we firstly constructed an E. coli cell surface display system for ZHD-P. The surface-displayed ZHD-P exhibited high activity against ZEN and showed optimal activity at 40 °C and pH 9.0. With superior pH stability, the surface-displayed ZHD-P retained 80% activity when it was incubated at pH 5.0-11.0 for 12 h. Interestingly, the metal ions tolerance of the surface-displayed ZHD-P was better than the intracellular form. Additionally, the surface-displayed ZHD-P could be reused four times with the residual enzyme activity of more than 50%. The biotoxicity assessment using P. phosphoreum T3 indicated that ZEN could be degraded into hypotoxic products by the intracellular or surface-displayed ZHD-P. ZHD-P could be feasible for ZEN detoxification.
Asunto(s)
Hidrolasas/genética , Proteínas Recombinantes/genética , Zearalenona/metabolismo , Secuencia de Aminoácidos , Escherichia coli , Regulación de la Expresión Génica , Concentración de Iones de Hidrógeno , Hidrolasas/metabolismo , Hypocreales , Unión Proteica , Proteínas Recombinantes/metabolismo , Propiedades de Superficie , Zearalenona/químicaRESUMEN
Zearalenone is a fungal contaminant that is widely present in grains. Here, a novel molecularly imprinted membrane based on SOM-ZIF-8 was developed for the rapid and highly selective identification of zearalenone in grain samples. The molecularly imprinted membrane was prepared using polyvinylidene fluoride, cyclododecyl 2,4-dihydroxybenzoate as a template and SOM-ZIF-8 as a carrier. The factors influencing the extraction of zearalenone using this membrane, including the solution pH, extraction time, elution solvent, elution time, and elution volume, were studied in detail. The optimized conditions were 5 mL of sample solution at pH 6, extraction time of 45 min, 4 mL of acetonitrile:methanol = 9:1 as elution solvent, and elution time of 20 min. This method displayed a good linear range of 12-120 ng/g (R2 = 0.998) with the limits of detection and quantification of this method are 1.7 and 5.5 ng/g, respectively. In addition, the membrane was used to selectively identify zearalenone in grain samples with percent recoveries ranging from 87.9 to 101.0% and relative standard deviation of less than 6.6%. Overall, this study presents a simple and effective chromatographic pretreatment method for detecting zearalenone in food samples.
Asunto(s)
Grano Comestible/química , Zearalenona/análisis , Cromatografía Líquida de Alta Presión/métodos , Industria Procesadora y de Extracción/métodos , Contaminación de Alimentos/análisis , Estructuras Metalorgánicas , Impresión Molecular/métodos , Polímeros Impresos Molecularmente , Micotoxinas/análisis , Micotoxinas/química , Extracción en Fase Sólida/métodos , Zearalenona/químicaRESUMEN
The design and fabrication of a surface-enhanced Raman scattering (SERS) aptasensor for simultaneous detection of zearalenone (ZEN) and ochratoxin A (OTA) in wheat and corn samples is described. The capture and reporter probes were SH-cDNA-modified gold nanorods and SH-Apt-modified Au@Ag core-shell nanoparticles, respectively. After recognizing OTA and ZEN aptamers and complementary strands (SH-cDNA), the reporter probe generated a strong SERS signal. The preferred binding of OTA and ZEN aptamers to OTA and ZEN, respectively, caused reporter probes to release the capture probes, resulting in a linear decrease in SERS intensity. The detection of OTA showed good linearity with an R2 value of 0.986, which could be maintained across a wide concentration range (0.01 to 100 ng/mL), with the limit of detection of 0.018 ng/mL. For detection of ZEN, good linearity with an R2 value of 0.987 could be maintained across a wide concentration range (0.05 to 500 ng/mL), with 0.054 ng/mL as the limit of detection. Good accuracy (relative standard deviation < 4.2%) during mycotoxin determination as well as excellent quantitative recoveries (96.0-110.7%) during the analysis of spiked real samples was achieved. The proposed SERS aptasensor exhibited excellent performance in the detection of OTA and ZEN in real food samples. Hence, by simply changing the aptamer, this new model can be applied to the detection of multiple mycotoxins in the food industry.
Asunto(s)
Técnicas Biosensibles/métodos , Nanopartículas del Metal/química , Micotoxinas/análisis , Nanotubos/química , Ocratoxinas/análisis , Zearalenona/análisis , Aptámeros de Nucleótidos/química , Grano Comestible/química , Contaminación de Alimentos/análisis , Oro/química , Ácidos Nucleicos Inmovilizados/química , Límite de Detección , Micotoxinas/química , Ocratoxinas/química , Reproducibilidad de los Resultados , Plata/química , Espectrometría Raman , Triticum/química , Zea mays/química , Zearalenona/químicaRESUMEN
Mycotoxins are toxic metabolites of filamentous fungi. Previous studies demonstrated the co-occurrence of Fusarium and Alternaria toxins, including zearalenone (ZEN), ZEN metabolites, and alternariol (AOH). These xenoestrogenic mycotoxins appear in soy-based meals and dietary supplements, resulting in the co-exposure to ZEN and AOH with the phytoestrogen genistein (GEN). In this study, the cytotoxic and estrogenic effects of ZEN, reduced ZEN metabolites, AOH, and GEN are examined to evaluate their individual and combined impacts. Our results demonstrate that reduced ZEN metabolites, AOH, and GEN can aggravate ZEN-induced toxicity; in addition, the compounds tested exerted mostly synergism or additive combined effects regarding cytotoxicity and/or estrogenicity. Therefore, these observations underline the importance and the considerable risk of mycotoxin co-exposure and the combined effects of mycoestrogens with phytoestrogens.
Asunto(s)
Estrógenos/toxicidad , Genisteína/toxicidad , Lactonas/toxicidad , Zearalenona/metabolismo , Zearalenona/toxicidad , Muerte Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Genisteína/química , Células HeLa , Humanos , Lactonas/química , Micotoxinas/toxicidad , Oxidación-Reducción , Zearalenona/químicaRESUMEN
Lactonohydrolase ZHD can detoxify oestrogenic mycotoxin zearalenone and zearalenols through hydrolysis and decarboxylation. The detail mechanism, especially the role of Trp183, which interacts with substrate through p-π interaction and one hydrogen bond, is still unknown. The Trp183 mutants abolished activity to ZEN, α-ZOL and ß-ZOL, except that W183F mutant retained about 40% activity against α-ZOL. In two W183F-reactant complex structures the reactants still bind at the active position and it suggested that this p-π interaction takes responsible for the reactants recognization and allocation. Further, the ZHD-productant complex structures showed that the resorcinol ring of hydrolysed α-ZOL and hydrolysed ß-ZOL move a distance of one ring as compare to the resorcinol ring of reactant α-ZOL and ß-ZOL. The same movement also found in comparison of hydrolysed ZEN and ZEN. In the structure of W183F complex with hydrolysed α-ZOL the resorcinol ring of hydrolysed α-ZOL doesn't move as compare to the resorcinol ring of reactant α-ZOL. It suggested the Trp183 coordinated hydrogen bond takes responsible for the movement of the hydrolysed product. These functional and structural results suggested that Trp183 is essential for ZHD detoxifying zearalenone and zearalenols.
Asunto(s)
Hidrolasas/metabolismo , Triptófano/química , Triptófano/metabolismo , Zearalenona/metabolismo , Zeranol/análogos & derivados , Biocatálisis , Hidrolasas/genética , Inactivación Metabólica , Modelos Moleculares , Proteínas Mutantes/química , Proteínas Mutantes/metabolismo , Mutación/genética , Relación Estructura-Actividad , Zearalenona/química , Zeranol/química , Zeranol/metabolismoRESUMEN
BACKGROUND: The root-knot nematode Meloidogyne graminicola has become a serious threat to rice production as a result of the cultivation changes from transplanting to direct seeding. The nematicidal activity of Aspergillus welwitschiae have been investigated in vitro, and the disease control efficacy of the active compound has been evaluated under greenhouse and field conditions. RESULTS: The active compound αß-dehydrocurvularin (αß-DC), isolated by nematicidal assay-directed fractionation, showed significant nematicidal activity against M. graminicola, with a median lethal concentration (LC50) value of 122.2 µg mL- 1. αß-DC effectively decreased the attraction of rice roots to nematodes and the infection of nematodes and also suppressed the development of nematodes under greenhouse conditions. Moreover, αß-DC efficiently reduced the root gall index under field conditions. CONCLUSIONS: To our knowledge, this is the first report to describe the nematicidal activity of αß-DC against M. graminicola. The results obtained under greenhouse and field conditions provide a basis for developing commercial formulations from αß-DC to control M. graminicola in the future.
Asunto(s)
Antiparasitarios/farmacología , Aspergillus/química , Oryza/crecimiento & desarrollo , Tylenchoidea/efectos de los fármacos , Zearalenona/análogos & derivados , Animales , Antiparasitarios/aislamiento & purificación , Línea Celular , Supervivencia Celular/efectos de los fármacos , Cromatografía , Femenino , Efecto Invernadero , Estructura Molecular , Oryza/parasitología , Enfermedades de las Plantas/prevención & control , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/parasitología , Tylenchoidea/crecimiento & desarrollo , Zearalenona/química , Zearalenona/aislamiento & purificación , Zearalenona/farmacologíaRESUMEN
The importance of transforming growth factor beta-activated kinase 1 (TAK1) to cell survival has been demonstrated in many studies. TAK1 regulates signalling cascades, the NF-κB pathway and the mitogen-activated protein kinase (MAPK) pathway. TAK1 inhibitors can induce the apoptosis of cancerous cells, and irreversible inhibitors such as (5Z)-7-oxozeaenol are highly potent. However, they can react non-specifically with cysteine residues in proteins, which may have serious adverse effects. Reversible covalent inhibitors have been suggested as alternatives. We synthesised imidazopyridine derivatives as novel TAK1 inhibitors, which have 2-cyanoacrylamide moiety that can form reversible covalent bonding. A derivative with 2-cyano-3-(6-methylpyridin-2-yl)acrylamide (13h) exhibited potent TAK1 inhibitory activity with an IC50 of 27 nM. It showed a reversible reaction with ß-mercaptoethanol, which supports its potential as a reversible covalent inhibitor.
Asunto(s)
Acrilamida/química , Imidazoles/síntesis química , Quinasas Quinasa Quinasa PAM/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/síntesis química , Piridinas/síntesis química , Sitios de Unión , Humanos , Imidazoles/metabolismo , Mercaptoetanol/química , Modelos Moleculares , FN-kappa B/metabolismo , Unión Proteica , Inhibidores de Proteínas Quinasas/metabolismo , Piridinas/metabolismo , Transducción de Señal , Relación Estructura-Actividad , Factor de Transcripción ReIA , Zearalenona/análogos & derivados , Zearalenona/químicaRESUMEN
An electrochemical aptasensor is described for determination of the phytohormone of zearalenone (ZEA). The gold electrode was modified with ZEA via covalent attachment using cysteamine-hydrochloride and 1,4-phenylene diisocyanate linker. A truncated ZEA aptamer with a dissociation constant of 13.4 ± 2.1 nM was used in an aptasensor. The electrochemical property was investigated using square wave voltammetry for monitoring the change in the electron transfer using the ferro/ferricyanide system as redox probe. Under optimal experimental conditions, the response was best measured at a potential of 0.20 V (vs. Ag/AgCl). The signals depended on the competitive mechanism between the immobilised ZEA and free ZEA for the aptamer binding site. The aptasensor works in the range 0.01 to 1000 ng·mL-1 ZEA concentration, with a detection limit of 0.017 ng·mL-1. High degree of cross-reactivity with the other analogues of ZEA was observed, whereas none towards other mycotoxins. The aptasensor was further applied for the determination of ZEA in the extract of maize grain and showed good recovery percentages between 87 and 110%. Graphical abstract Schematic representation of the electrochemical determination of zearalenone based on indirect competitive assay. Step a Immobilisation of ZEA on the surface of gold electrode via covalent attachment, b competition for the ZEA aptamer binding site between immobilised and free ZEA, and c current signal of the binding event based on SWV technique.
Asunto(s)
Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Micotoxinas/análisis , Zearalenona/análisis , Secuencia de Bases , Técnicas Electroquímicas/instrumentación , Electrodos , Contaminación de Alimentos/análisis , Oro/química , Ácidos Nucleicos Inmovilizados/química , Límite de Detección , Micotoxinas/química , Zea mays/química , Zearalenona/químicaRESUMEN
An aptamer-based assay for the determination of two different kinds of fusarium mycotoxins, i.e., zearalenone (ZEN) and fumonisin B1 (FB1), is presented. Based on the inner filter effect (IFE) strategy, the contents of ZEN and FB1 can be simultaneously quantified. It is making use of 65-nm gold nanorods (AuNRs), 20-nm upconversion nanoparticles (UCNPs), fluorescence dyes, and DNA sequences. In the absence of ZEN and FB1, the UCNPs and AuNRs associate through DNA sequences. Due to IFE effect, weak fluorescence signals are collected. In the presence of ZEN or FB1, UCNPs and AuNRs become unstable and partially separate from each other. This results in the recovery of fluorescence signals. Under 980-nm laser excitation, the logarithmic values of fluorescence signal intensities at 606 nm and 753 nm gradually increase with the concentration of ZEN and FB1 in the ranges 0.05-100 µg L-1 (the coefficient of determination is 0.997) and 0.01-100 ng L-1 (the coefficient of determination is 0.986), respectively. The limits of detection (LOD) of the fabricated assay for ZEN and FB1 are 0.01 µg L-1 and 0.003 ng L-1, respectively. The proposed method has a high selectivity over other competitive mycotoxins, including aflatoxin B1, ochratoxin A, patulin and ochratoxin B. The applicability of the assay was evaluated in the determination of ZEN and FB1 contents in spiked corn samples. The average recoveries ranged from 89.9 to 106.6%. This result confirms the practicality of this method. Graphical abstract Schematic representation of an aptamer-based fluorometric method for simultaneous determination of two kinds of the fusarium mycotoxins zearalenone and fumonisin B1.
Asunto(s)
Aptámeros de Nucleótidos/química , Fumonisinas/análisis , Nanopartículas del Metal/química , Micotoxinas/análisis , Nanotubos/química , Zearalenona/análisis , Colorantes Fluorescentes/química , Contaminación de Alimentos/análisis , Fumonisinas/química , Oro/química , Límite de Detección , Micotoxinas/química , Espectrometría de Fluorescencia , Zea mays/química , Zearalenona/químicaRESUMEN
An aptasensor is reported for the detection of three different kinds of mycotoxins, i.e., zearalenone (ZEN), ochratoxin A (OTA), and fumonisin B1 (FB1). Based on fluorescence resonance energy transfer effect (FRET) and surface-enhanced Raman scattering (SERS), the levels of ZEN, FB1, and OTA can be simultaneously determined. Under 980-nm and 650-nm laser excitation, the logarithmic values of fluorescence signal intensities at 543 nm and 670 nm are slowly increased as the concentrations of ZEN and OTA vary from 0.1 ng mL-1 and 0.05 ng mL-1 to 100 ng mL-1 and 25 ng mL-1, respectively. For FB1, under 980-nm laser excitation, the logarithmic value of SERS signal intensity at 1567 cm-1 gradually increases with the concentration of FB1 in the range 0.05-200 pg mL-1 (R2 = 0.996). The detection limits of the proposed assay for ZEN, OTA, and FB1 are 0.03 ng mL-1, 0.01 ng mL-1, and 0.02 pg mL-1, respectively. The selectivity experiment results indicate this assay possesses a high selectivity over other commonly encountered mycotoxins. The average recoveries range from 90 to 107%, revealing satisfactory application potential of the proposed assay. The developed aptasensor will bring bright prospects for research in the field of multiplexed mycotoxine detection. Graphical Abstract Schematic representation of an aptamer-based assay for multiple mycotoxins determination.
Asunto(s)
Aptámeros de Nucleótidos/química , Transferencia Resonante de Energía de Fluorescencia/métodos , Micotoxinas/análisis , Espectrometría Raman/métodos , Carbocianinas/química , ADN/química , Colorantes Fluorescentes/química , Contaminación de Alimentos/análisis , Fumonisinas/análisis , Fumonisinas/química , Oro/química , Límite de Detección , Nanopartículas del Metal/química , Micotoxinas/química , Conformación de Ácido Nucleico , Ocratoxinas/análisis , Ocratoxinas/química , Zea mays/química , Zearalenona/análisis , Zearalenona/químicaRESUMEN
The development of a bioluminescent immunosensor is reported for the determination of zearalenone (ZEA) based on a peptide mimetic identified by phage display. The peptide mimetic GW, with a peptide sequence GWWGPYGEIELL, was used to create recombinant fusion proteins with the bioluminescent Gaussia luciferase (GLuc) that were directly used as tracers for toxin detection in a competitive immunoassay without the need for secondary antibodies or further labeling. The bioluminescent sensor, based on protein G-coupled magnetic beads for antibody immobilization, enabled determination of ZEA with a detection limit of 4.2 ng mL-1 (corresponding to 420 µg kg-1 in food samples) and an IC50 value of 11.0 ng mL-1. The sensor performance was evaluated in spiked maize and wheat samples, with recoveries ranging from 87 to 106% (RSD < 20%, n = 3). Finally, the developed method was applied to the analysis of a naturally contaminated reference matrix material and good agreement with the reported concentrations was obtained.Graphical abstract.
Asunto(s)
Peptidomiméticos/química , Proteínas Recombinantes de Fusión/química , Zearalenona/químicaRESUMEN
The co-occurrence of moniliformin (MON), fumonisins (FBs), and deoxynivalenol (DON) was evaluated in maize, durum, and common wheat grown in different experimental fields located in several Italian regions. MON was quantified using a LC-MS/MS method adding lanthanum ions in the mobile phase. In maize, MON contamination was widespread and considerable; the toxin was detected in almost all the samples (95.1%) and exceeded 500 and 1000 µg kg-1 in 42.0% and in 18.5% of samples, respectively. Significant positive correlation was found between MON and FB contamination levels. When there were not droughty climate conditions, a positive significant correlation was found between growing degree days (GDD) and MON values. In wheat, MON contamination was not widespread like in maize and it was lower in common wheat than in durum wheat. In durum wheat, MON was detected in 45.0% of the samples with only 6 samples (7.5%) exceeding 500 µg kg-1, while in common wheat the toxin was detected above the LOD in 18.7% of samples exceeding 100 µg kg-1 in only two samples (2.5%). No correlation was found with DON contamination. Climate conditions influenced both MON and DON occurrence.
Asunto(s)
Ciclobutanos/química , Contaminación de Alimentos , Micotoxinas/química , Toxina T-2/química , Ciclobutanos/aislamiento & purificación , Grano Comestible/química , Fusarium/química , Fusarium/patogenicidad , Humanos , Italia , Micotoxinas/aislamiento & purificación , Toxina T-2/aislamiento & purificación , Espectrometría de Masas en Tándem , Triticum/química , Triticum/crecimiento & desarrollo , Triticum/microbiología , Zea mays/química , Zea mays/crecimiento & desarrollo , Zea mays/microbiología , Zearalenona/química , Zearalenona/aislamiento & purificaciónRESUMEN
Activation of synovial fibroblasts (SFs) contributes to rheumatoid arthritis (RA) by damaging synovial membranes and generating inflammatory cytokines that recruit immune cells to the joint. In this paper we profile cytokine secretion by primary human SFs from healthy tissues and from donors with RA and show that SF activation by TNF, IL-1α, and polyinosinic-polycytidylic acid (Poly(I:C)) cause secretion of multiple cytokines found at high levels in RA synovial fluids. We used interaction multiple linear regression to quantify therapeutic and countertherapeutic drug effects across activators and donors and found that the ability of drugs to block SF activation was strongly dependent on the identity of the activating cytokine. (5z)-7-oxozeaenol (5ZO), a preclinical drug that targets transforming growth factor-ß-activated kinase 1 (TAK1), was more effective at blocking SF activation across all contexts than the approved drug tofacitinib, which supports the development of molecules similar to 5ZO for use as RA therapeutics.
Asunto(s)
Antirreumáticos/farmacología , Artritis Reumatoide/tratamiento farmacológico , Fibroblastos/efectos de los fármacos , Líquido Sinovial/citología , Membrana Sinovial/efectos de los fármacos , Zearalenona/análogos & derivados , Antirreumáticos/química , Artritis Reumatoide/patología , Células Cultivadas , Citocinas/biosíntesis , Humanos , Modelos Lineales , Membrana Sinovial/metabolismo , Zearalenona/química , Zearalenona/farmacologíaRESUMEN
A unified and concise approach to the synthesis of nine curvularin-type metabolites and two analogues has been developed with few steps and high yields. Among them, sumalactones A-D were synthesized for the first time. The key steps in this approach included esterification, Friedel-Crafts acylation, and ring-closing metathesis (or cross metathesis).