RESUMO
Herein, we synthesized anemone-like copper-based metal-organic frameworks (MOFs) loaded with gold-palladium nanoparticles (AuPd@Cu-MOFs) and polyethylenimine-reduced graphene oxide/gold-silver nanosheet composites (PEI-rGO/AuAg NSs) for the first time to construct the sensor and to detect T-2 toxin (T-2) using triple helix molecular switch (THMS) and signal amplification by swing-arm robot. The aptasensor used PEI-rGO/hexagonal AuAg NSs as the electrode modification materials and anemone-like AuPd@Cu-MOFs as the signal materials. The prepared PEI-rGO/hexagonal AuAg NSs had a large specific surface area, excellent electrical conductivity, and good stability, which successfully improved the electrochemical performance of the sensors. The AuPd@Cu-MOFs with high porosity provided a great deal of attachment sites for the signaling molecule thionine (Thi), thereby increasing the signal response. The aptasensor developed in this study demonstrated a remarkable detection limit of 0.054 fg mL-1 under optimized conditions. Furthermore, the successful detection of T-2 in real samples was achieved using the fabricated sensor. The simplicity of the THMS-based method, which entails modifying the aptamer sequence, allows for easy adaptation to different target analytes. Thus, the sensor holds immense potential for applications in quality supervision and food safety.
Assuntos
Anemone , Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Estruturas Metalorgânicas , Robótica , Toxina T-2 , Estruturas Metalorgânicas/química , Cobre/química , Nanopartículas Metálicas/química , Aptâmeros de Nucleotídeos/química , Paládio , Grafite/química , Ouro/química , Técnicas Eletroquímicas/métodos , Limite de Detecção , Técnicas Biossensoriais/métodosRESUMO
Based on DNA strand replacement reaction and aptamer-specific recognition, a simple dual-labeled DNA nanostructure is designed for the simultaneous detection of Ochratoxin A (OTA) and aflatoxin B1 (AFB1). C1 is labeled with Cy3 and Cy5, while C2 and C3 are labeled with BHQ2. The fluorescence intensity of DNA nanostructure composed of C1, C2 and C3 is weak because of fluorescence resonance energy transfer. When OTA Aptamer (OTA-Apt) and AFB1 Aptamer (AFB1-Apt) are added to the homogeneous system at the same time, C1 can be replaced with the help of toehold strand displacement, resulting in fluorescence enhancement. In the presence of both OTA and AFB1, the toehold strand displacement reaction is inhibited due to preferential binding between the target and their corresponding aptamers. The limit of detection of OTA was 0.007 ng/mL and that of AFB1 was 0.03 ng/mL. The recoveries of OTA and AFB1 were 96%-101% and 97%-101% in the corn sample, and 99%-101% and 92%-106% in the wine sample. Compared with other sensors, the preparation of this aptasensor needs simpler experimental steps and a shorter total-preparing time, confirming the convenient, rapid, and time-saving operation process.
Assuntos
Nanoestruturas , Aflatoxina B1/química , DNA/química , Corantes Fluorescentes/química , Limite de DetecçãoRESUMO
Developing rapid detection technology for adenosine triphosphate (ATP) is crucial in quality supervision and food safety. Herein, an electrochemical aptasensor based on an aptazyme-catalyzed signal amplification strategy is constructed for ATP detection using polyethyleneimine-functionalized molybdenum disulfide (PEI-MoS2)/Au@PtPd nanobipyramids (MoS2/Au@PtPd NBPs) as a modification material. Additionally, a novel kind of nitrogen-rich covalent organic framework (COF) is prepared using melamine and cyanuric acid (MCA). We synthesize MCA and the Co-based metal organic framework (Co-MOF) as the signal label. Due to the fact that π-π stacking interactions of Co-MOF@MCA can expand the load efficiency and surface concentration of the signal label, the signal response is an order of magnitude higher than that of Co-MOF or MCA as the signal label. Target ATP changes the conformation of the aptazyme, and it becomes activated. With the assistance of metal ions, the signal label is circularly cleaved, causing an amplification of the signal. Among them, MoS2/Au@PtPd NBPs have a large specific surface area and good electrical conductivity and can carry substantial DNA strands and amplify the redox signal of methylene blue (MB). Under optimal conditions, the aptasensor can detect ATP from 10 pM to 100 µM with a low limit of detection of 7.37 × 10-10 µM. Therefore, the novel aptasensor has extensive application prospects in quality supervision and food safety.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Estruturas Metalorgânicas , Nanoestruturas , Trifosfato de Adenosina , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas , Ouro/química , Limite de Detecção , Nanopartículas Metálicas/química , Estruturas Metalorgânicas/química , Azul de Metileno , Molibdênio/química , Nanoestruturas/química , Nitrogênio , Polietilenoimina , TriazinasRESUMO
As a common heavy metal ion with strong toxicity and wide distribution, lead ions (Pb2+) had great harm to the human body. In this work, a low-noise ratiometric fluorescence biosensor was developed based on Pb2+-dependent DNAzyme and exonuclease III (Exo III)-assisted cascade signal amplification. Firstly, the substrate chain of DNAzyme (S-DNA) was modified on the surface of magnetic beads (MBs) through the combination of biotin and streptavidin, and then the enzyme chain of DNAzyme (E-DNA) was connected to the MBs by forming a double-stranded DNA (dsDNA) with S-DNA. A hairpin DNA (HP) labelled with Cy3 and Cy5 respectively at both ends was used as a fluorescence probe. The emission peaks of Cy3 and Cy5 can appear at 562 nm and 665 nm respectively, and their fluorescence intensity ratio (F562/F665) was chosen as the acquisition signal. The ratiometric sensor can reduce the interference of detection environment and avoid false positive reactivity. Due to the cleavage of DNAzyme and the release of single-stranded DNA (ssDNA) in the presence of Pb2+, the hairpin structure of HP was opened and the FRET between two fluorophores disappeared, resulting in the strengthened signal of Cy3 and the weakened signal of Cy5. Furthermore, the ratio [Formula: see text] signal increased gradually with the increase of Pb2+ concentration. When the concentration of Pb2+ was in the range of 0.1-1000 nM, [Formula: see text] had a good linear relationship with [Formula: see text], the correlation coefficient (R2) was 0.997, and the limit of detection (LOD) was 77 pM. The presented ratiometric fluorescence biosensor had lower LOD and wider detection range via comparing with other methods. At the same time, the sensor also obtained the satisfactory results for detection of Pb2+ in tap water, tea, and rice flour samples. The provided ratiometric biosensor has great potential in the monitoring of various targets. A low-noise ratiometric fluorescence biosensor based on the FRET between two fluorophores was developed, and the DNAzyme and exonuclease III-assisted cascade signal amplification was used to improve the sensitivity of the method. The biosensor had a detection limit as low as 77 pM.
Assuntos
DNA Catalítico/metabolismo , Exodesoxirribonucleases/metabolismo , Chumbo/análise , Técnicas Biossensoriais/métodos , Fluorescência , HumanosRESUMO
The simultaneous detection of multiple mycotoxins is of great significance for food safety and human health. Herein, a simple, convenient and accurate fluorescent aptasensor was designed based on the dual cross DNA nanostructure for the simultaneous detection of ochratoxin A (OTA) and aflatoxin B1 (AFB1), in which the stable dual cross DNA nanostructure provided an assay platform using the fluorescent dye-labeled aptamers as a sensing element. Owing to the higher affinity of aptamers for their target, the aptamer probes were released from the assay platform in the presence of OTA and AFB1, resulting in an enhanced fluorescence at 570 nm and 670 nm. This "signal-on" fluorescent aptasensor assay system can effectively avoid background signals and minimize false positive. Furthermore, the designed method can realize the simultaneous detection of OTA and AFB1 during the whole experiment. The limits of detection (LOD) were as low as 0.0058 ng/mL for OTA, ranging from 0.01 to 50 ng/mL and 0.046 ng/mL for AFB1, ranging from 0.05 to 100 ng/mL. The proposed fluorescent aptasensor exhibits excellent performance in practical application and provides a novel approach for the simultaneous detection of multiple mycotoxins by simply changing the aptamers. A "signal-on" fluorescent aptasensor assay system based on the stable dual cross DNA nanostructure was successfully developed for simultaneous detection of OTA and AFB1 with lower detection limits in wider linear ranges.
Assuntos
Aflatoxina B1/análise , Aptâmeros de Nucleotídeos/química , DNA/química , Corantes Fluorescentes/química , Conformação de Ácido Nucleico , Ocratoxinas/análise , Eletroforese em Gel de Ágar , Limite de Detecção , Reprodutibilidade dos TestesRESUMO
The presented voltammetric mercury(II) sensor is based on the specific interaction between Hg(II) ion and thymine-thymine base pairs. Reduced graphene oxide is functionalized with gold nanorods and then loaded with thionine and streptavidin (RGO@AuNR-TH-SA). A T-rich thiolated DNA (S1) is firstly immobilized on a gold electrode. In the presence of Hg (II), the T-rich biotin-DNA (biotin-S2) binds to S1 via T-Hg(II)-T interaction. Then, the RGO@AuNR-TH-SA is linked to the gold electrode by specific binding between SA and biotin-S2. This produces an electrochemical signal (at -0.208 V vs. Ag/AgCl) of TH that depends on the concentration of Hg (II). The peak current increases linearly in the 1 to 200 nM Hg (II) concentration range, and the detection limit is 0.24 nM. The sensor is highly selective for Hg (II) over other environmentally relevant metal ions, even at concentration ratios of >1000. Graphical abstract Schematic representation of a voltammetric biosensor for mercury(II) using reduced graphene oxide@gold nanorods (RGO@AuNRs) and thymine-Hg(II)-thymine interaction. It is based on the fact that RGO@AuNR can strongly adsorb thionine (TH) and streptavidin to realize the signal amplification.
Assuntos
Técnicas Biossensoriais/métodos , DNA/química , Grafite/química , Mercúrio/análise , Nanotubos/química , Timina/química , Pareamento de Bases , DNA/genética , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Ouro/química , Ácidos Nucleicos Imobilizados/química , Limite de Detecção , Fenotiazinas/química , Reprodutibilidade dos Testes , Estreptavidina/química , Poluentes Químicos da Água/análiseRESUMO
Peanut meal is the byproduct of high-temperature peanut oil extraction; it is mainly composed of proteins, which have complex tastes after enzymatic hydrolysis to free amino acids and small peptides. The enzymatic hydrolysis method was adopted by using two compound proteases of trypsin and flavorzyme to hydrolyze peanut meal aiming to provide a flavor base. Hence, it is necessary to assess the taste attributes and assign definite taste scores of peanut meal double enzymatic hydrolysis hydrolysates (DEH). Conventionally, sensory analysis is used to assess taste intensity in DEH. However, it has disadvantages because it is expensive and laborious. Hence, in this study, both taste attributes and taste scores of peanut meal DEH were evaluated using an electronic tongue. In this regard, the response characteristics of the electronic tongue to the DEH samples and standard five taste samples were researched to qualitatively assess the taste attributes using PCA and DFA. PLS and RBF neural network (RBFNN) quantitative prediction models were employed to compare predictive abilities and to correlate results obtained from the electronic tongue and sensory analysis, respectively. The results showed that all prediction models had good correlations between the predicted scores from electronic tongue and those obtained from sensory analysis. The PLS and RBFNN prediction models constructed using the voltage response values from the sensors exhibited higher correlation and prediction ability than that of principal components. As compared with the taste performance by PLS model, that of RBFNN models was better. This study exhibits potential advantages and a concise objective taste assessment tool using the electronic tongue in the assessment of DEH taste attributes in the food industry.
Assuntos
Arachis/química , Arachis/classificação , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Nariz Eletrônico , Nozes/química , Algoritmos , Análise por Conglomerados , Hidrólise , Modelos LinearesRESUMO
In this study, an application of a voltammetric electronic tongue for discrimination and prediction of different varieties of rice was investigated. Different pretreatment methods were selected, which were subsequently used for the discrimination of different varieties of rice and prediction of unknown rice samples. To this aim, a voltammetric array of sensors based on metallic electrodes was used as the sensing part. The different samples were analyzed by cyclic voltammetry with two sample-pretreatment methods. Discriminant Factorial Analysis was used to visualize the different categories of rice samples; however, radial basis function (RBF) artificial neural network with leave-one-out cross-validation method was employed for prediction modeling. The collected signal data were first compressed employing fast Fourier transform (FFT) and then significant features were extracted from the voltammetric signals. The experimental results indicated that the sample solutions obtained by the non-crushed pretreatment method could efficiently meet the effect of discrimination and recognition. The satisfactory prediction results of voltammetric electronic tongue based on RBF artificial neural network were obtained with less than five-fold dilution of the sample solution. The main objective of this study was to develop primary research on the application of an electronic tongue system for the discrimination and prediction of solid foods and provide an objective assessment tool for the food industry.
Assuntos
Técnicas Eletroquímicas/instrumentação , Eletrônica/instrumentação , Oryza/química , Análise Discriminante , Eletricidade , Eletrodos , Análise Fatorial , Análise de Fourier , Redes Neurais de Computação , Soluções , Fatores de Tempo , LínguaRESUMO
IFN-ß is a critical antiviral cytokine that is capable of modulating the systemic immune response. The transcriptional induction of IFN-ß is a highly regulated process, involving the activation of pattern recognition receptors and their downstream signaling pathways. The Akt family of serine/threonine kinases includes three isoforms. The specific role for the individual Akt isoforms in pattern recognition and signaling remains unclear. In this article, we report that the TLR3-mediated expression of IFN-ß is blunted in cells that lack Akt1. The expression of IFN-ß-inducible genes such as CCL5 and CXCL10 was also reduced in Akt1-deficient cells; the induction of TNF-α and CXCL2, whose expression does not rely on IFN-ß, was not reduced in the absence of Akt1. Macrophages from Akt1(-/-) mice displayed deficient clearance of HSV-1 along with reduced IFN-ß expression. Our results demonstrate that Akt1 signals through ß-catenin by phosphorylation on Ser(552), a site that differs from the glycogen synthase kinase 3 ß phosphorylation site. Stimulation of a chemically activated version of Akt1, in the absence of other TLR3-dependent signaling, was sufficient for accumulation and phosphorylation of ß-catenin at Ser(552). Taken together, these results demonstrate that the Akt1 isoform is required for ß-catenin-mediated promotion of IFN-ß transcription downstream of TLR3 activation.
Assuntos
Interferon beta/fisiologia , Proteínas Proto-Oncogênicas c-akt/fisiologia , Transcrição Gênica/imunologia , beta Catenina/metabolismo , Animais , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fosforilação/imunologia , Isoformas de Proteínas/deficiência , Isoformas de Proteínas/genética , Isoformas de Proteínas/fisiologia , Proteínas Proto-Oncogênicas c-akt/deficiência , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Receptor 3 Toll-Like/metabolismo , beta Catenina/fisiologiaRESUMO
Lateral flow immunoassay (LFIA) has been employed extensively for the rapid, accurate, and portable detection of foodborne toxins. Here, the platinum gold nanoflower core-shell (Pt@AuNF) nanozyme with excellent optical properties, good catalytic ability and controllable reaction conditions were prepared to effectively improve the performance of lateral flow immunoassay (LFIA) strips. The Pt@AuNF nanozyme and horseradish peroxidase (HRP) combined with monoclonal antibody were used as signal probes based on the dual enzymes catalytic signal amplification strategy to detect Zearalenone sensitively. Dual enzymes catalyze the decomposition of hydrogen peroxide into hydroxyl radicals, and under the influence of hydroxyl radicals, colorless 3,3',5,5' -tetramethylbenzidine (TMB) is oxidized to blue ox-TMB, which is superimposed on the strips for signal amplification to broaden the detection range. The limit of detection (LOD) of the Pt@AuNF-HRP labeled LFIA strips after signal amplification was 0.052 ng/mL, and the detection range was 0.052-7.21 ng/mL. Compared with the Pt@AuNF labeled strips, while reducing the probes amount by half to achieve antibody conservation, the detection range was expanded by 5-fold based on achieving improved sensitivity. The study provided a meaningful reference for expanding the detection range based on immunoassay.
Assuntos
Nanopartículas Metálicas , Zearalenona , Peroxidase do Rábano Silvestre , Limite de Detecção , Imunoensaio , OuroRESUMO
Herein, a study for the first application of a hybridization chain reaction, a 1,8-naphthalimides-DNA (NDs) intercalator, and DNA-dependent Prussian blue nanoflowers@PtPd materials (PBNFs@PtPd) in the development of a fluorescence-electrochemical (FL-EC) aptasensor. This construction establishes an efficient sensing platform for the detection of procymidone (PCM). In the context of the described experiment, dual-mode detection is achieved through the generation of FL signals by an aptamer labeled with a Cy5 moiety and the formation of DPV signals by the modification of a thionine-appended 1,8-naphthalimide (Thi-NDs). In the presence of PCM, specific recognition occurs, followed by the utilization of magnetic separation technology to release DNA1 (S1) and aptamer-Cy5 (Apt-Cy5), subsequently introducing them onto both fluorescence and EC platforms. The presence of S1 effectively activates hybridization chain reaction (HCR) for the electrode surface, thereby significantly increasing the binding sites for Thi-NDs and consequently greatly amplifying the response signal of differential pulse voltammetry (DPV). The developed FL-EC dual-mode sensing platform demonstrates high sensitivity in the detection of PCM, with the detection limits of 0.173 µg·ml-1 (within the detection range of 500 pg·ml-1 to 500 ng·ml-1) and 0.074 ng·ml-1 (within the detection range of 100 pg·ml-1 to 100 ng·ml-1), respectively. The designed dual-mode sensor exhibits notable characteristics, including high selectivity, reproducibility, synergy, and reliable monitoring/capability for PCM in real samples.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Eletroquímicas , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , DNA/análise , Fluorescência , Hibridização de Ácido Nucleico , Poluentes Químicos da Água/análise , Limite de DetecçãoRESUMO
Balancing the accuracy and simplicity of aptasensors is a challenge in their construction. This study addresses this issue by leveraging the remarkable loading capacity and peroxidase-like catalytic activity of PtPdCu trimetallic nanoparticles, which reduces the reliance on precious metals. A dual-signal readout aptasensor for enrofloxacin (ENR) detection is designed, incorporating DNA dynamic network cascade reactions to further amplify the output signal. Exploiting the strong loading capacity of PtPdCu nanoparticles, they are self-assembled with thionine (Thi) to form a signal label capable of generating signals in two independent modes. The label exhibits excellent enzyme-like catalytic activity and enhances electron transfer capabilities. Differential pulse voltammetry (DPV) and square-wave voltammetry (SWV) are employed to independently read signals from the oxidation-reduction reaction of Thi and the catalytic oxidation of hydroquinone (HQ) to benzoquinone (BQ) by H2O2. The introduced DNA dynamic network cascade reaction modularizes sample processing and electrode surface signal generation, avoiding electrode contamination and efficiently increasing the output of the catalyzed hairpin assembly (CHA) cycle. Under optimized conditions, the developed aptasensor demonstrates detection limits of 0.112 (DPV mode) and 0.0203 pg/mL (SWV mode). Additionally, the sensor successfully detected enrofloxacin in real samples, expanding avenues for designing dual-mode signal amplification strategies.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Cobre , Enrofloxacina , Nanopartículas Metálicas , Platina , Enrofloxacina/análise , Aptâmeros de Nucleotídeos/química , Nanopartículas Metálicas/química , Técnicas Biossensoriais/métodos , Cobre/química , Platina/química , Rutênio/química , Técnicas Eletroquímicas/métodos , Limite de Detecção , Oxirredução , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/análise , Catálise , Antibacterianos/análise , Antibacterianos/químicaRESUMO
Heavy metal pollution in the environment has become a significant global concern due to its detrimental effects on human health and the environment. In this study, we report an electrochemical aptasensor for the simultaneous detection of Hg2+ and Pb2+. Gold nanoflower/polyethyleneimine-reduced graphene oxide (AuNFs/PEI-rGO) was introduced on the surface of a gold electrode to improve sensing performance. The aptasensor is based on the formation of a T-Hg2+-T mismatch structure and specific cleavage of the Pb2+-dependent DNAzyme, resulting in a dual signal generated by the Exo III specific digestion of methylene blue (MB) labeled at the 3' end of probe DNA-1 and the reduction of the substrate ascorbic acid (AA) catalyzed by the signal label. The decrease of MB signal and the increase of AA oxidation peak was used to indicate the content of Hg2+ and Pb2+, respectively, with detection limits of 0.11 pM (Hg2+) and 0.093 pM (Pb2+). The aptasensor was also used for detecting Hg2+ and Pb2+ in water samples with good recoveries. Overall, this electrochemical aptasensor shows promising potential for sensitive and selective detection of heavy metals in environmental samples.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Exodesoxirribonucleases , Chumbo , Mercúrio , Estruturas Metalorgânicas , Poluentes Químicos da Água , Mercúrio/análise , Chumbo/análise , Chumbo/química , Estruturas Metalorgânicas/química , Aptâmeros de Nucleotídeos/química , Exodesoxirribonucleases/química , Exodesoxirribonucleases/metabolismo , Poluentes Químicos da Água/análise , Técnicas Biossensoriais/métodos , Grafite/química , Ouro/química , Limite de Detecção , Eletrodos , DNA Catalítico/químicaRESUMO
Herein, an aptasensor based on a signal amplification strategy was developed for the sensitive detection of procymidone (PCM). AgPd nanoparticles/Polenimine Graphite oxide (AgPdNPs/PEI-GO) was weaned as electrode modification material to facilitate electron transport and increase the active sites on the electrode surface. Besides, Pt@Ni-Co nanoboxes (Pt@Ni-CoHNBs) were utilized to be carriers for signaling tags, after hollowing ZIF-67 and growing Pt, the resulting Pt@Ni-CoHNBs has a tremendous amounts of folds occurred on the surface, enables it to carry a larger quantity of thionine, thus amplify the detectable electrochemical signal. In the presence of PCM, the binding of PCM to the signal probe would trigger a change in electrical signal. The aptasensor was demonstrated with excellent sensitivity and a low detection limit of 0.98 pg·mL-1, along with a wide linear range of 1 µg·mL-1 to 1 pg·mL-1. Meanwhile, the specificity, stability and reproducibility of the constructed aptasensor were proved to be satisfactory.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Grafite , Limite de Detecção , Nanopartículas Metálicas , Paládio , Platina , Prata , Grafite/química , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Platina/química , Técnicas Biossensoriais/métodos , Nanopartículas Metálicas/química , Paládio/química , Prata/química , Níquel/química , Polietilenoimina/química , Cobalto/química , Reprodutibilidade dos TestesRESUMO
Lead ion pollution has become a serious public health concern worldwide. Therefore, sensitive detection of Pb2+ is critical to control lead pollution, assess risks, and safeguard the health of vulnerable populations. This study reports a highly sensitive labelling-free electrochemical aptasensor for Pb2+ detection. The aptasensor employs silver-platinum nanoparticles/graphene oxide (AgPt/GO) and Exonuclease III (Exo III) for signal amplification. GO provides high surface area and conductivity for immobilizing AgPt NPs, facilitating the immobilization of aptamer (Apt) probes on the electrode surface. Exo III enzymatically cleaves DNA strands on the electrode surface, releasing DNA segments to amplify the signal further. The synergistic amplification by AgPt/GO and ExoIII enables an extremely wide linear detection range of 0.05 pM-5 nM for Pb2+, with a low detection limit of 0.019 pM. Additionally, the G-quadruplex structure ensures excellent selectivity for Pb2+ detection, resulting in high reproducibility and stability of the aptasensor. The aptasensor was successfully applied to detect spiked Pb2+ in tap water samples, achieving recovery rates ranging from 96 to 108.4 %. By integrating nanomaterials, aptamers and enzymatic amplification, the aptasensor facilitates highly sensitive and selective detection of Pb2+, demonstrating potential for practical applications in environmental monitoring.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Exodesoxirribonucleases , Grafite , Chumbo , Nanocompostos , Platina , Prata , Grafite/química , Chumbo/análise , Chumbo/química , Aptâmeros de Nucleotídeos/química , Exodesoxirribonucleases/química , Técnicas Eletroquímicas/métodos , Platina/química , Nanocompostos/química , Prata/química , Técnicas Biossensoriais/métodos , Nanopartículas Metálicas/química , Limite de Detecção , Poluentes Químicos da Água/análise , Água Potável/análise , Eletrodos , Quadruplex GRESUMO
Citrinin (CIT) is a mycotoxin with nephrotoxicity and hepatotoxicity, presenting a significant threat to human health that is often overlooked. Therefore, a dual-signal mode (DPV and SWV) aptasensor for citrinin (CIT) detection was constructed based on tetrahedral DNA nanostructures (TDN) in this study. Furthermore, PtPdCo mesoporous nanozymes exhibit catalase-like catalytic functions, generating significant electrochemical signals through a Fenton-like reaction. Meanwhile their excellent Methylene Blue (MB) loading capability ensures independent dual signal outputs. The RecJf exonuclease-assisted (RecJf Exo-assisted) process can expand the linear detection range, enabling further amplification of the signal. Under optimized conditions, the constructed aptaensor exhibited excellent detection performance with limits of detection (LODs) of 7.67 × 10-3 ng·mL-1 (DPV mode) and 1.57 × 10-3 ng·mL-1 (SWV mode). Due to its multiple signal amplification and highly accurate dual-signal mode detection capability, this aptasensor shows promising potential for the in situ detection.
RESUMO
The γ(1)34.5 protein of herpes simplex viruses (HSV) is essential for virulence. Accordingly, an HSV mutant lacking γ(1)34.5 is attenuated in vivo. Despite its vaccine potential, the mechanism by which the γ(1)34.5 null mutant triggers protective immunity is unknown. In this report we show that vaccination with the γ(1)34.5 null mutant protects against lethal challenge from wild-type virus via IκB kinase in dendritic cells (DCs), which sense virus-associated molecular patterns. Unlike mock-treated DCs, DCs primed with the γ(1)34.5 null mutant ex vivo mediate resistance to wild-type HSV after adoptive transfer into naïve mice. Furthermore, the γ(1)34.5 null mutant activates IκB kinase, which facilitates p65/RelA phosphorylation and nuclear translocation, resulting in DC maturation. While unable to produce infectious virus in DCs, this mutant virus expresses early and late genes. In its abortive infection, the γ(1)34.5 null mutant induces protective immunity more effectively in CD8(+) DCs than in CD8(-) DCs. This is mirrored by a higher level of interleukin-6 (IL-6) and IL-12 secretion by CD8(+) DCs than CD8(-) DCs. Remarkably, inhibition of p65/RelA phosphorylation or nuclear translocation in CD8(+) DCs disrupts protective immunity. These results suggest that engagement of the γ(1)34.5 null mutant with CD8(+) DCs elicits innate immunity to activate NF-κB, which translates into protective immunity.
Assuntos
Células Dendríticas/imunologia , Herpes Simples/imunologia , Herpesvirus Humano 1/imunologia , NF-kappa B/imunologia , Deleção de Sequência , Proteínas Virais/genética , Animais , Antígenos CD8/genética , Antígenos CD8/imunologia , Células Cultivadas , Células Dendríticas/virologia , Herpes Simples/genética , Herpes Simples/virologia , Humanos , Imunidade , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/genética , Proteínas Virais/imunologiaRESUMO
The γ(1)34.5 protein of herpes simplex viruses (HSV) is essential for viral pathogenesis, where it precludes translational arrest mediated by double-stranded-RNA-dependent protein kinase (PKR). Paradoxically, inhibition of PKR alone is not sufficient for HSV to exhibit viral virulence. Here we report that γ(1)34.5 inhibits TANK binding kinase 1 (TBK1) through its amino-terminal sequences, which facilitates viral replication and neuroinvasion. Compared to wild-type virus, the γ(1)34.5 mutant lacking the amino terminus induces stronger antiviral immunity. This parallels a defect of γ(1)34.5 for interacting with TBK1 and reducing phosphorylation of interferon (IFN) regulatory factor 3. This activity is independent of PKR. Although resistant to IFN treatment, the γ(1)34.5 amino-terminal deletion mutant replicates at an intermediate level between replication of wild-type virus and that of the γ(1)34.5 null mutant in TBK1(+/+) cells. However, such impaired viral growth is not observed in TBK1(-/-) cells, indicating that the interaction of γ(1)34.5 with TBK1 dictates HSV infection. Upon corneal infection, this mutant replicates transiently but barely invades the trigeminal ganglia or brain, which is a difference from wild-type virus and the γ(1)34.5 null mutant. Therefore, in addition to PKR, γ(1)34.5 negatively regulates TBK1, which contributes viral replication and spread in vivo.
Assuntos
Regulação para Baixo , Herpes Simples/virologia , Herpesvirus Humano 1/fisiologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Virais/metabolismo , Replicação Viral , Animais , Linhagem Celular , Herpesvirus Humano 1/genética , Humanos , Fator Regulador 3 de Interferon/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Fosforilação , Ligação Proteica , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Virais/genéticaRESUMO
Improving urban resilience (UR) and enhancing urban anti-risk ability are important foundations for promoting the high-quality development of new urbanization. This research employs the time-varying entropy method to evaluate the resilience level of 138 cities within China's eight urban agglomerations (UAs) between 2005 and 2019. Additionally, the Dagum Gini coefficient and the kernel density estimation method are utilized to examine the spatial disparities and distribution dynamics of UR across the eight UAs. The results of this investigation indicate that (1) the collective UR performance of the eight UAs has experienced an upward trend. However, a notable spatial disparity exits, which is primarily attributed to the differences among the UAs. (2) The overall UR development of the eight UAs has a certain gradient effect, and the UR within each UA has different degrees of polarization characteristics. (3) For the eight UAs as a whole, per capita savings deposits, capitalization of foreign capital, and per capita fiscal expenditure are the three most important driving factors. Within each UA, there was heterogeneity in the main influencing factors. The interplay between any two factors amplifies their individual driving effects on the spatial differentiation of UR.
RESUMO
Mycotoxins contaminated in agricultural products are often highly carcinogenic and genotoxic to humans. With the streamlining of the food industry chain and the improvement of food safety requirements, the traditional laboratory testing mode is constantly challenged due to the expensive equipment, complex operation steps, and lag in testing results. Therefore, rapid detection methods are urgently needed in the food safety system. This review focuses on the latest strategies that can achieve rapid and on-site testing, with particular attention to the nanomaterials integrated biosensors. To provide researchers with the latest trends and inspiration in the field of rapid detection, we summarize several strategies suitable for point of care testing (POCT) of mycotoxins, including enzyme-linked immunoassay (ELISA), lateral flow assay (LFA), fluorescence, electrochemistry, and colorimetry assay. POCT-based strategies are all developing towards intelligence and portability, especially when combined with smartphones, making it easier to read signals for intuitive access and analysis of test data. Detection performance of the devices has also improved considerably with the integration of biosensors and nanomaterials.