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Antimicrobial resistance has become a serious threat to the global public health. Accurate and rapid antimicrobial susceptibility testing (AST) allows evidence-based prescribing of antibiotics to improve patient care and clinical outcomes. Current culture-based AST assays are inherently limited by the doubling time of bacterial reproduction, which require at least 24 h to have a decisive result. Herein, a label-free electrical impedance-based microfluidic platform designed to expedite and streamline AST procedure for clinical practice is presented. Following a 30-min exposure of bacterial samples to antibiotics, the presented high-throughput, single-bacterium level impedance characterization platform enables a rapid 2-min AST assay. The platform facilitates accurate analysis of individual bacterial viability, as indicated by changes in electrical characteristics, thereby enabling the determination of antimicrobial resistance. Moreover, the potential clinical applicability of this platform is demonstrated by testing different E. coli strains against five antibiotics, yielding 100% categorical agreements compared to standard culture methods.
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Escherichia coli , Microfluídica , Humanos , Impedância Elétrica , Testes de Sensibilidade Microbiana , Antibacterianos/farmacologia , BactériasRESUMO
This study investigated the impact of wheat processing methods (wheat flour vs wheat pellets) on the growth performance, serum biochemical parameters, and rumen microbiome composition in sheep. Results indicated that feeding of wheat flour resulted in significantly higher terminal weight and average daily gain (P < 0.05) and lower cholesterol and ALP04 levels (P < 0.05) in sheep compared to those fed wheat pellets. Analysis of 16s rDNA high-throughput sequencing data revealed significantly higher microbial richness (Chao1 index) in the rumen of sheep fed wheat flour (P < 0.05), even though the phylum-level composition dominated by Firmicutes, Bacteroidetes, and Proteobacteria was similar in both groups of sheep. Notably, sheep fed wheat flour were found to have a significantly higher relative abundance of Bacteroidetes (P < 0.05). At the genus level, Succinivibrionaceae_UCG-001 and Prevotella_1 were significantly more abundant in the rumen of sheep fed wheat flour (P < 0.05). Correlation analysis identified that both terminal weight and average daily gain were positively correlated with ruminal abundance of Bacteroidetes and Prevotella_1, while ALP04 was negatively correlated with the abundance of these taxa. Functional prediction using PICRUSt2 indicated enrichment of pathways related to the ABC-type glycerol-3-phosphate transport system, and periplasmic components in both wheat flour and pellet fed sheep. Overall, these findings suggest that dietary wheat flour modulates rumen microbiota composition, and improves growth performance in sheep.
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For critically ill patients with non-small cell lung cancer (NSCLC) in need of life-saving treatment, there is currently no reported evidence regarding the use of medication specifically targeting epidermal growth factor receptor ( EGFR ) p.C797S mutation, which is known to cause resistance to third-generation tyrosine kinase inhibitors (TKIs). Our report aims to investigate and explore treatment strategies to overcome resistance associated with EGFR p.C797S mutation in order to provide potential therapeutic options for these patients. Here, we reported two cases with NSCLC who initially harbored an EGFR -sensitive mutation and were both treated with osimertinib, a third-generation TKI. Next-generation sequencing tests conducted prior to the initiation of fifth-line therapy in critically ill patients revealed the presence of EGFR p.C797S mutations in both patients, suggesting acquired resistance. In the course of fifth-line therapy, the administration of a combination of brigatinib and cetuximab proved vital in saving critically ill patients, moderately extending their overall survival period. Our findings suggested that a combined regimen of brigatinib and cetuximab could serve as a potentially life-saving therapeutic strategy for critically ill patients with NSCLC, particularly those demonstrating EGFR p.C797S-mediated resistance. Further studies, however, are required to validate and expand upon these promising findings.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica , Carcinoma Pulmonar de Células não Pequenas , Cetuximab , Receptores ErbB , Neoplasias Pulmonares , Mutação , Compostos Organofosforados , Pirimidinas , Humanos , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Receptores ErbB/genética , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Pirimidinas/uso terapêutico , Pirimidinas/administração & dosagem , Cetuximab/administração & dosagem , Cetuximab/uso terapêutico , Masculino , Compostos Organofosforados/uso terapêutico , Compostos Organofosforados/administração & dosagem , Pessoa de Meia-Idade , Feminino , Estado Terminal , Idoso , Resistencia a Medicamentos Antineoplásicos , Acrilamidas/uso terapêutico , Compostos de Anilina , IndóisRESUMO
Stimulant laxatives were recently found to be abused in slimming foods, resulting in harmful effects on consumers. To ensure the safety of relative products, sensitive yet multiplex immunoassays are crucial in rapid screening of stimulant laxatives. However, there are few immunoassays for these substances, and even less for broad-specific recognition. Thus, in this work, four theoretically promising haptens of emerging stimulant laxative bisacodyl were rationally designed using molecular modeling and synthesized to immune animals, whose feasibility was confirmed by the obtained broad-specific antibody. Based on this unique antibody, a highly sensitive multiplex competitive indirect enzyme-linked immunosorbent assay (ciELISA) was established with low limits of detection for bisacodyl, sodium picosulfate, and BHPM (0.23, 13.68, and 0.11 ng/mL). In spiked sample recovery test and real sample detection, this ciELISA exhibited acceptable consistency with the validation method, demonstrating high accuracy and applicability of our method. This reliable multiplex ciELISA proceeds the rapid screening of stimulant laxatives in slimming foods.
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Ensaio de Imunoadsorção Enzimática , Laxantes , Ensaio de Imunoadsorção Enzimática/métodos , Laxantes/análise , Limite de Detecção , Contaminação de Alimentos/análise , Animais , Anticorpos/imunologia , Análise de Alimentos/métodos , Haptenos/química , Haptenos/imunologiaRESUMO
A novel enantioselective Tsuji-Trost-type cross coupling reaction between gem-difluorinated cyclopropanes and N-unprotected amino acid esters enabled by synergistic Pd/Ni/chiral aldehyde catalysis is presented herein. This transformation streamlined the diversity-oriented synthesis (DOS) of optically active α-quaternary α-amino acid esters bearing a linear 2-fluoroallylic motif, which served as an appealing platform for the construction of other valuable enantioenriched compounds. The key intermediates were confirmed by HRMS detection, while DFT calculations revealed that the excellent enantioselectivity was attributed to the stabilizing non-covalent interactions between the Pd(II)-π-fluoroallyl species and the Ni(II)-Schiff base complex.
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The widespread use of sulfonamide (SA) antibiotics in animal husbandry has led to residues of SAs in the environment, causing adverse effects to the ecosystem and a risk of bacterial resistance, which is a potential threat to public health. Therefore, it is highly desirable to develop simple, high-throughput methods that can detect multiple SAs simultaneously. In this study, we isolated aptamers with different specificities based on a multi-SA systematic evolution of ligands by the exponential enrichment (SELEX) strategy using a mixture of sulfadimethoxine (SDM), sulfaquinoxaline (SQX), and sulfamethoxazole (SMZ). Three aptamers were obtained, and one of them showed a similar binding to all tested SAs, with dissociation constant (Kd) ranging from 0.22 to 0.63 µM. For the other two aptamers, one is specific for SQX, and the other is specific for SDM and sulfaclozine. A label-free detection method based on the broad-specificity aptamer was developed for the simultaneous detection of six SAs, with detection of limits ranging from 0.14 to 0.71 µM in a lake water sample. The aptasensor has no binding for other broad-spectrum antibiotics such as ß-lactam antibiotics, quinolones, tetracyclines, and chloramphenicol. This work provides a promising biosensor for rapid, multiresidue, and high-throughput detection of SAs, as well as a shortcut for the preparation of different specific recognition elements required for the detection of broad-spectrum antibiotics.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Animais , Antibacterianos , Aptâmeros de Nucleotídeos/química , Ecossistema , Sulfanilamida , Sulfadimetoxina , Sulfonamidas , Sulfaquinoxalina , Técnicas Biossensoriais/métodos , Técnica de Seleção de Aptâmeros/métodosRESUMO
BACKGROUND: Primary biliary cholangitis (PBC) is a chronic cholestatic liver disease characterized by inflammation of the interlobular bile ducts. Ursodeoxycholic acid (UDCA) is the only FDA approved first-line therapy for PBC, but up to 40% of patients with PBC have an incomplete response to UDCA. Neutrophil-to-lymphocyte (NLR) has been used to predict prognosis in various liver diseases. There is limited evidence on the treatment response to UDCA in PBC patients. Our study aimed to evaluate the relationship between NRL and the response to UDCA treatment in PBC patients. METHODS: A total of 257 primary biliary cholangitis (PBC) patients treated with UDCA (13-15 mg/kg/d) were enrolled in this retrospective study. The response to treatment was evaluated based on alkaline phosphatase levels ≤1.67 times the upper limit of the normal value after 12 months of UDCA treatment. Multivariable logistic regression analysis was performed to investigate the association between NLR at baseline and the response to 12 months of UDCA treatment after adjusting for important confounding variables. The stability of the results was evaluated by unadjusted and adjusted models. RESULTS: The results of multiple regression analysis showed that NLR at baseline was positively associated with the nonresponse to UDCA treatment after adjustments for potential confounders (age, sex, BMI, hypertension, arterial plaque, thyroid disease, jaundice, albumin, globulin, total bile acid, ALP, GGT, LDLC, total cholesterol, hemoglobin, and APTT) (OR = 1.370, 95% CI 1.066-1.761). These results reveal that NLR is an independent risk factor for UDCA treatment nonresponse. CONCLUSIONS: Our results suggest that PBC patients with a high NLR had a worse response to UDCA therapy.
Assuntos
Cirrose Hepática Biliar , Ácido Ursodesoxicólico , Humanos , Ácido Ursodesoxicólico/uso terapêutico , Ácido Ursodesoxicólico/efeitos adversos , Estudos Retrospectivos , Cirrose Hepática Biliar/tratamento farmacológico , Cirrose Hepática Biliar/complicações , Colagogos e Coleréticos/uso terapêutico , Neutrófilos , Resultado do TratamentoRESUMO
Propisochlor is a chloroacetamide herbicide causing liver toxicity and suppressing immunity in human and animal. Although the herbicide has been used for years, the effects of propisochlor on intestinal health remain poorly understood. Hence, the impacts of propisochlor in intestinal health and gut microbiota were analyzed by using molecular approach and bacterial 16S rRNA sequencing. The result showed that the intake of propisochlor in mice impaired gut morphology, reduced expression of tight junction proteins, decreased thickness of mucus layer and activated pyroptosis signaling. Moreover, the exposure of propisochlor in mice led to significant alterations in gut microbial diversity and composition, including an increase of Bacteroidetes and a decrease of Firmicutes. The gut microbiota, such as Parabacteroides, Parasutterella, and Bacteroides, demonstrated a strong negative correlation with the intestinal health. These findings suggested that gut microbiota could play a critical role in the propisochlor-induced pyroptosis.
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Mycotoxin contamination in foods and other goods has become a broad issue owing to serious toxicity, tremendous threat to public safety, and terrible loss of resources. Herein, it is necessary to develop simple, sensitive, inexpensive, and rapid platforms for the detection of mycotoxins. Currently, the limitation of instrumental and chemical methods cannot be massively applied in practice. Immunoassays are considered one of the best candidates for toxin detection due to their simplicity, rapidness, and cost-effectiveness. Especially, the field of dual-mode immunosensors and corresponding assays is rapidly developing as an advanced and intersected technology. So, this review summarized the types and detection principles of single-mode immunosensors including optical and electrical immunosensors in recent years, then focused on developing dual-mode immunosensors including integrated immunosensors and combined immunosensors to detect mycotoxins, as well as the combination of dual-mode immunosensors with a portable device for point-of-care test. The remaining challenges were discussed with the aim of stimulating future development of dual-mode immunosensors to accelerate the transformation of scientific laboratory technologies into easy-to-operate and rapid detection platforms.
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Técnicas Biossensoriais , Micotoxinas , Micotoxinas/análise , Técnicas Biossensoriais/métodos , Imunoensaio/métodos , AlimentosRESUMO
The detection of rosiglitazone (RSG) in food is of great importance since the excessive intake of RSG could cause adverse effects on the human body. Although liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry are the preliminary methods for the detection of hazardous materials in food, they are not suitable for point-of-care or on-site detection. Herein, a time-based readout (TBR) device with an application software (APP) controlled by a smart phone was developed for the sensitive and selective immunoassay of RSG. The homemade TBR device was based on a two-electrode system, where the immune molecule-modified glassy carbon electrode was used as the bioanode, and Prussian blue-modified FTO was used as the cathode. By using Au-modified octahedral Cu2O with high catalytic activity as mimetic peroxidase, an insulating layer was generated on the cathode by catalyzing 4-chloro-1-naphthol (4-CN) into benzo-4-chlorohexadienone (B4Q). The time to reach a fixed potential varied indirectly with the concentrations of RSG and was recognized by the APP, while the electrochromic property on the cathode was also correspondingly changed. Under optimum conditions, both the square root of the time and the chroma value of the electrochromism exhibited linear responses for the detection of RSG ranging from 5 × 10-10 to 5 × 10-7 g/L, while the limits of detection were 8.2 × 10-11 and 1.3 × 10-10 g/L, respectively. With easy operation and portability, this TBR device showed a promising application for point-of-care monitoring of hazardous materials in food or the environment.
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Técnicas Biossensoriais , Dioxigenase FTO Dependente de alfa-Cetoglutarato , Eletrodos , Substâncias Perigosas , Humanos , Imunoensaio , RosiglitazonaRESUMO
Developing an easily-prepared, sensitive, and accurate point-of-need immunochromatographic assay (ICA) is significant in food safety screening, clinical diagnosis, and environmental monitoring. However, the current single-modal ICAs are limited in certain instinct drawbacks that restrict analytical performances. Herein, we introduce an ultrasensitive dual-modal colorimetric/reversed ratiometric fluorescence ICA based on facilely prepared immunoprobes with a high loading capacity of red quantum dots and AuNPs. By smartly integrating these red-colored/fluorescent signal probes with an immobilized green quantum dot antigen on the test lines, discrete "turn-on" visual inspection and reversed ratiometric quantification via a portable smartphone-based analyzer were accomplished. As an application, this method was employed to detect 11 phosphodiesterase-5 inhibitors in health foods with ultralow detection limits (0.0028-0.045 ng/mL), high repeatability (coefficient of variations of 0.3-1.91%), and reasonable accuracy (recoveries of 86.6-107%). The proposed method was further validated by the authorized liquid chromatography with tandem mass spectrometry method in actual sample detection. This new assay format can be extended to ultrasensitive flexible detection of other food contaminants, environmental pollutants, or tumor biomarkers within minutes, and it just requires simply prepared signal reporters, easy-to-operate procedures, and a low-cost miniaturized analyzer.
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Poluentes Ambientais , Nanopartículas Metálicas , Pontos Quânticos , Biomarcadores Tumorais , Colorimetria , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5 , Corantes Fluorescentes , Ouro/química , Imunoensaio/métodos , Limite de Detecção , Nanopartículas Metálicas/química , Pontos Quânticos/químicaRESUMO
Recently, necroptosis has emerged as one of the important mechanisms of ischemia stroke. Necroptosis can be rapidly activated in endothelial cells to cause vascular damage and neuroinflammation. Panax notoginseng saponins (PNS), an ingredient extracted from the root of Panax notoginseng (Burk.) F.H. Chen, was commonly used for ischemic stroke, while its molecular mechanism and targets have not been fully clarified. Our study aimed to clarify the anti-necroptosis effect of PNS by regulating RIP1-RIP3-MLKL signaling pathway in brain microvascular endothelial cells (BMECs) subjected to transient oxygen-glucose deprivation (OGD/resupply [R]). In vitro, the necroptosis model of rat BMECs was established by testing the effect of OGD/R in the presence of the pan-caspase inhibitor z-VAD-FMK. After administration of PNS and Nec-1, cell viability, cell death modality, the expression of RIP1-RIP3-MLKL pathway and mitochondrial membrane potential (Δψm) level were investigated in BMECs upon OGD/R injury. The results showed that PNS significantly enhanced cell viability of BMECs determined by CCK-8 analysis, and protected BMECs from necroptosis by Flow cytometry and TEM. In addition, PNS inhibited the phosphorylation of RIP1, RIP3, MLKL and the downstream expression of PGAM5 and Drp1, while similar results were observed in Nec-1 intervention. We further investigated whether PNS prevented the Δψm depolarization. Our current findings showed that PNS effectively reduced the occurrence of necroptosis in BMECs exposed to OGD/R by inhibition of the RIP1-RIP3-MLK signaling pathway and mitigation of mitochondrial damage. This study provided a novel insight of PNS application in clinics.
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Panax notoginseng , Saponinas , Animais , Encéfalo/metabolismo , Caspases/metabolismo , Caspases/farmacologia , Células Endoteliais/metabolismo , Glucose/metabolismo , Necroptose , Oxigênio/metabolismo , Panax notoginseng/química , Proteínas Quinases/metabolismo , Ratos , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Saponinas/farmacologia , Transdução de SinaisRESUMO
With the aggravation and evolution of global warming, natural disasters such as hurricanes occur more frequently, posing a great challenge to large-scale power systems. Therefore, the pre-position and reconfiguration of the microgrid defense resources by means of Mobile Energy Storage Vehicles (MEVs) and tie lines in damaged scenarios have attracted more and more attention. This paper proposes a novel two-stage optimization model with the consideration of MEVs and tie lines to minimize the shed loads and the outage duration of loads according to their proportional priorities. In the first stage, tie lines addition and MEVs pre-position are decided prior to a natural disaster; in the second stage, the switches of tie lines and original lines are operated and MEVs are allocated from staging locations to allocation nodes according to the specific damaged scenarios after the natural disaster strikes. The proposed load restoration method exploits the benefits of MEVs and ties lines by microgrid formation to pick up more critical loads. The progressive hedging algorithm is employed to solve the proposed scenario-based two-stage stochastic optimization problem. Finally, the effectiveness and superiority of the proposed model and applied algorithm are validated on an IEEE 33-bus test case.
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Desastres Naturais , AlgoritmosRESUMO
Food safety analysis plays a significant role in controlling food contamination and supervision. In recent years, multiplex optical bioassays (MOBAs) have been widely applied to analyze multiple hazards due to their efficiency and low cost. However, due to the challenges such as multiplexing capacity, poor sensitivity, and bulky instrumentation, the further application of traditional MOBAs in food screening has been limited. In this review, effective strategies regarding food safety MOBAs are summarized, such as spatial-resolution modes performed in multi-T lines/dots strips or arrays of strip/microplate/microfluidic chip/SPR chip and signal-resolution modes employing distinguishable colorimetric/luminescence/fluorescence/surface plasma resonance/surface-enhanced Raman spectrum as signal tags. Following this, new trends on how to design engineered sensor architecture and exploit distinguishable signal reporters, how to improve both multiplexing capacity and sensitivity, and how to integrate these formats into smartphones so as to be mobile are summarized systematically. Typically, in the case of enhancing multiplexing capacity and detection throughput, microfluidic array chips with multichannel architecture would be a favorable approach to overcome the spatial and physical limitations of immunochromatographic assay (ICA) test strips. Moreover, noble metal nanoparticles and single-excitation, multiple-emission luminescence nanomaterials hold great potential in developing ultrasensitive MOBAs. Finally, the exploitation of innovative multiplexing strategy hybridized with powerful and widely available smartphones opens new perspectives to MOBAs. In future, the MOBAs should be more sensitive, have higher multiplexing capacity, and easier instrumentation.
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Bioensaio , Contaminação de Alimentos/análise , Inocuidade dos Alimentos , Imunoensaio , Análise Espectral RamanRESUMO
Food fraud is currently a growing global concern with far-reaching consequences. Food authenticity attributes, including biological identity, geographical origin, agricultural production, and processing technology, are susceptible to food fraud. Metabolic markers and their corresponding authentication methods are considered as a promising choice for food authentication. However, few metabolic markers were available to develop robust analytical methods for food authentication in routine control. Untargeted metabolomics by liquid chromatography-mass spectrometry (LC-MS) is increasingly used to discover metabolic markers. This review summarizes the general workflow, recent applications, advantages, advances, limitations, and future needs of untargeted metabolomics by LC-MS for identifying metabolic markers in food authentication. In conclusion, untargeted metabolomics by LC-MS shows great efficiency to discover the metabolic markers for the authenticity assessment of biological identity, geographical origin, agricultural production, processing technology, freshness, cause of animals' death, and so on, through three main steps, namely, data acquisition, biomarker discovery, and biomarker validation. The application prospects of the selected markers by untargeted metabolomics require to be valued, and the selected markers need to be eventually applicable at targeted analysis assessing the authenticity of unknown food samples.
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Metabolômica , Espectrometria de Massas em Tandem , Animais , Biomarcadores/análise , Cromatografia Líquida , Alimentos , Metabolômica/métodos , Espectrometria de Massas em Tandem/métodosRESUMO
The abnormal expression of microRNA (miRNA) can affect the RNA transcription and protein translation, leading to tumor progression and metastasis. Currently, the accurate detection of aberrant expression of miRNA, particularly using a portable detection system, remains a great challenge. Herein, a novel dual-mode biosensor with high sensitivity and robustness for miR-21 detection was developed based on the cis-cleavage and trans-cleavage activities of Cas12a. miRNA can be combined with hairpin DNA-horseradish peroxidase anchored on a CdS/g-C3N4/B-TiO2 photoelectrode, thus the nonenzymatic amplification was triggered to form numerous HRP-modified double-stranded DNA (HRP-dsDNA). Then, HRP-dsDNA can be specifically recognized and efficiently cis-cleaved by Cas12a nucleases to detach HRP from the substrate, while the remaining HRP on HRP-dsDNA can catalyze 4-chloro-1-naphthol (4-CN) to form biocatalytic precipitation (BCP) on the surface of the photoelectrode, and thus the photocurrent can be changed. Meanwhile, the trans-cleavage ability of Cas12a was activated, and nonspecifically degrade the FQ-reporter and a significant fluorescence signal can be generated. Such two different kinds of signals with independent transmission paths can mutually support to improve the performance of the detection platform. Besides, a portable device was constructed for the point-of-care (POC) detection of miR-21. Moreover, the dual-mode detection platform can be easily expanded for the specific detection of other types of biomarkers by changing the sequence of hairpin DNA, thereby promoting the establishment of POC detection for early cancer diagnosis.
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Técnicas Biossensoriais , MicroRNAs , Sistemas CRISPR-Cas , DNA , Peroxidase do Rábano Silvestre , MicroRNAs/genéticaRESUMO
Three kinds of immunochromatographic assays (ICAs) are proposed for the highly sensitive and rapid determination of tylosin (TYL) and tilmicosin (TIM) in eggs based on colloidal gold (CG), latex microsphere (LM), and time-resolved fluorescent microsphere (TRFM). Three types of ICAs could tolerate the egg matrix via simple sample pretreatment and demonstrated high sensitivity for TYL and TIM with cut-off values of 6/6/3 µg/kg and 14/14/6 µg/kg, respectively. Furthermore, in a single-blind parallel study 20 egg samples were analyzed by the three developed ICAs and confirmed by LC-MS/MS. The results showed good consistency, and there were no false positive and false negative results in our three ICAs. Consequently, the proposed three ICAs offered rapid, highly sensitive, reliable, and selectable testing platforms for screening veterinary medicine or other small molecule contaminants.
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Antibacterianos/análise , Cromatografia de Afinidade/métodos , Ovos/análise , Contaminação de Alimentos/análise , Tilosina/análogos & derivados , Tilosina/análise , Animais , Coloide de Ouro/química , Látex/química , Microesferas , Nanopartículas/química , Método Simples-CegoRESUMO
Establishing rapid, simple, and in situ detection of microcystin-LR (MC-LR) in drinking water sources is of significant importance for human health. To ease the situation that current methods cannot address, an open surface droplet microfluidic magnetosensor was designed and validated to quantify MC-LR in reservoir water, which is capable of (1) MC-LR isolation via MC-LR antibody-conjugated magnetic beads, (2) parallel and multistep analytical procedures in 15-array power-free and reusable active droplet microfluidic chips, (3) immunoassay incubation and fluorescence excitation within a miniaturized multifunctional 3D-printing optosensing accessory, and (4) signal read-out and data analysis by a user-friendly Android app. The proposed smartphone-based fluorimetric magnetosensor exhibited a low limit of detection of 1.2 × 10-5 µg/L in the range of 10-4 µg/L to 100 µg/L. This integrated and high throughput platform was utilized to draw an MC-LR contamination map for six reservoirs distributed in the Pearl River delta, Guangdong Province. It promises to be a simple and successful quantification method for MC-LR field detection, bringing many benefits to rapid on-site screening.
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Água Potável/química , Toxinas Marinhas/análise , Microcistinas/análise , Técnicas Analíticas Microfluídicas , Fluorometria , Campos Magnéticos , Tamanho da Partícula , Smartphone , Software , Propriedades de SuperfícieRESUMO
Tetrabromobisphenol A (TBBPA) is the largest brominated flame retardant which can be released to environment and cause long-term hazard. In this work, we developed a rapid and highly sensitive fluorescence enzyme-linked immunosorbent assay (FELISA) for monitoring of TBBPA in soil samples. TBBPA specific nanobody derived from camelid was fused with alkaline phosphatase to obtain the bi-functional fusion protein, which enable the specific binding of TBBPA and the generation of detection signal simultaneously. The assay showed an IC50 of 0.23â¯ngâ¯g-1, limit detection of 0.05â¯ngâ¯g-1 and linear range from 0.1 to 0.55â¯ngâ¯g-1 for TBBPA in soil samples. Due to the high resistance to organic solvents of the fusion protein, a simple pre-treatment by using 40% dimethyl sulfoxide (DMSO) as extract solvent can eliminate matrix effect and obtain good recoveries (ranging from 93.4% to 112.4%) for spiked soil samples. Good relationship between the results of the proposed FELISA and that of liquid chromatography tandem mass spectrometry (LC-MS/MS) was obtained, which indicated it could be a powerful analytical tool for determination of TBBPA to monitor human and environmental exposure.
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Monitoramento Ambiental/métodos , Ensaio de Imunoadsorção Enzimática , Retardadores de Chama/análise , Bifenil Polibromatos/análise , Poluentes do Solo/análise , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Camelídeos Americanos , Limite de Detecção , Bifenil Polibromatos/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Anticorpos de Domínio Único/genética , Anticorpos de Domínio Único/metabolismoRESUMO
A series of haptens were rationally designed for producing monoclonal antibodies specific for EC and a simple fluorescence immunoassay platform was developed for the sensitive determination of EC based on alkaline phosphatase (ALP)-triggered Cu+ quenching of CdSe quantum dots (QDs). It was noted that Cd as a fluorescence substrate in CdSe QDs can be selectively substituted by Cu+ that resulted in a more significant fluorescence quenching in comparison with Cu2+. Meanwhile, because ALP catalyzed ascorbic acid phosphate and then assisted the transformation of Cu2+ to Cu+, the change in fluorescence intensity was found to be proportional to ALP concentration. After simple magnetic separation, the sensitivity and linear range of the established assay were improved approximately 53-fold and an order of magnitude, respectively, when compared with the conventional ELISA. The proposed platform was able to both amplify the signal and eliminate matrix interferences, making it a promising to determine EC as well as other contaminants in complex food matrix in a highly sensitive and simple manner. Graphical abstract.