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The public health concern of antibiotic residues in animal-origin food has been a long-standing issue. In this work, we present a novel method for antibiotic detection, leveraging optical weak value amplification and harnessing an indirect competitive inhibition assay, which significantly boosts the system's sensitivity in identifying small molecule antibiotics. We chose chloramphenicol as a model compound and mixed it with chloramphenicol-bovine serum albumin conjugates to bind to the chloramphenicol antibody competitively. We achieved a broad linear detection range of up to 3.24 ng/mL and a high concentration resolution of 33.20 pg/mL. To further validate the universality of our proposed detection methodology, we successfully applied it to testing gibberellin and tetracycline. Moreover, we conducted regeneration experiments and real-sample correlation studies. This study introduces a novel strategy for the label-free optical sensing of small molecule antibiotics, greatly expanding the range of applications for sensors utilizing optical weak value amplification.
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Allergy is a prevalent disease, and the potential allergic population is expanding with industrialization and changes in people's living standards. Serum immunoglobulin E (IgE) level is one of the critical indicators for determining allergy. Here, we proposed a simple, real-time monitoring, low chip cost, label-free aptamer biosensing strategy based on weak value amplification (WVA) for the quantitative detection of IgE in serum samples, enabling early and accurate diagnosis of allergic or hypersensitive patients. The aptasensor combined an imaging weak measurement system with the high specificity of the aptamer for the marker IgE. By modifying the amino group at the 3-terminal end, the anti-IgE aptamers can attach to a dopamine-modified prism's surface and selectively recognize IgE in human serum. In the presence of IgE, a specific binding reaction occurred, resulting in a change in the refractive index of the reactive region's surface, manifested as a change in the light intensity of the camera acquired experimental images. As the concentration of IgE increased, the relative light intensity advanced sequentially. The WVA-aptasensing strategy achieved a wide detection range of 0.01 ng/mL to 2 µg/mL in phosphate buffered saline buffer, with the resolution as low as 4.3 pg/mL. IgE testing experiments in human serum have proved the feasibility of our methods in detecting complex samples. In addition, the method specifically recognized IgE without interference from other proteins. We believe that our proposed sensing strategy opens up new possibilities for ultrahigh sensitivity screening of IgE and can be expanded to detecting other biomolecules.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Imunoglobulina E , Imunoglobulina E/sangue , Humanos , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Limite de DetecçãoRESUMO
A label-free optical sandwich immunoassay sensor, utilizing weak value amplification and total internal reflection, was devised for real-time, high-sensitivity analysis and detection of low-concentration targets. 3D printed channels and sodium chloride solution were employed to ensure reproducibility, reliability, and stability of the measurements for calibration. The sandwich structure demonstrated enhanced responsiveness in the proposed optical biosensor through a comparative analysis of the direct assay and sandwich assay for detecting alpha-fetoprotein (AFP) at the same concentration. By optimizing the binding sequences of the coating antibody, target, and detection antibody in the sandwich method, a more suitable sandwich sensing approach based on weak value amplification was achieved. With this approach, the limit of detection (LOD) of 6.29 ng/mL (pM level) for AFP in PBS solution was achieved. AFP testing and regeneration experiments in human serum have proved the feasibility of our methods in detecting complex samples and the reusability of sensing chips. Additionally, the method demonstrated excellent selectivity for unpaired antigens. The efficacy of this methodology was evaluated by simultaneously detecting AFP, carcinoembryonic antigen (CEA), and CA15-3 on a singular sensor chip. In conclusion, the label-free sandwich immunoassay sensing scheme holds promise for advancing the proposed optical sensors based on weak value amplification in early diagnosis and prevention applications. Compared to other biomarker detection methods, it will be easier to promote in practical applications.
Assuntos
Técnicas Biossensoriais , Antígeno Carcinoembrionário , Limite de Detecção , alfa-Fetoproteínas , Técnicas Biossensoriais/métodos , alfa-Fetoproteínas/análise , Humanos , Antígeno Carcinoembrionário/sangue , Antígeno Carcinoembrionário/análise , Imunoensaio/métodos , Mucina-1/sangue , Mucina-1/análise , Anticorpos Imobilizados/imunologia , Anticorpos Imobilizados/químicaRESUMO
Five new diisoprenyl cyclohexene-type meroterpenoids, aspergienynes J-N (1-5), along with three known analogues (6-8), were obtained from the mangrove endophytic fungal strain Aspergillus sp. GXNU-Y85. The chemical structures, including their absolute configurations, were established via spectroscopic data and comparison of experimental and calculated ECD spectra. Cytotoxicity assay results indicated that compound 8 had strong cytotoxicity against HeLa cancer cells, and its IC50 value was 11.8 µM. In addition, flow cytometry analysis revealed that the cytotoxicity of 8 was due to the induction of G1 cell cycle arrest and apoptosis in HeLa cells.
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Antineoplásicos , Aspergillus , Humanos , Estrutura Molecular , Células HeLa , Aspergillus/química , Análise Espectral , Antineoplásicos/farmacologia , Antineoplásicos/metabolismoRESUMO
A new benzoquinone, guxiumasperone A (1), and a new diisoprenyl-cyclohexene-type meroterpenoids, biscognienyne M (2), along with four known diisoprenyl-cyclohexene analogues was isolated from the mangrove endophytic fungus Aspergillus QG1a. Their structures were determined by extensive spectral analysis of 1D and 2D NMR, HR-ESI- MS, and X-ray crystallography. Compound 1 was deduced by a single-crystal X-ray diffraction analysis, and the absolute configuration of 2 was further unequivocally elucidated by comparing the experimental electronic circular dichroism (ECD) data with calculated ECD spectra. Compounds 1 and 2 showed significant cytotoxic activity against selected tumour cells. Particularly, compound 2 exhibited strong activity against A2780 cancer cells with an IC50 value of 6.8 µM.
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A new sulphur-containing metabolite, asperiguxidione A (1), was isolated from a mangrove endophytic fungus Aspergillus sp. GXNU-MA, and four known alkaloids 2-5, were isolated together from this strain. Their structures were determined by the com-bination of 1D and 2D NMR spectroscopy, HR-ESI-MS, and ECD analysis. Compounds 1 and 2 exhibited mediate activity against Staphylococcus aureus and Enterobacter aerogenes with equal MIC values of 12.5 µg/mL. Compound 3 reduced NO production in LPS-stimulated cells with an IC50 value of 13.329 ± 0.53 µg/mL in the anti-inflammatory assay.
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An investigation on bioactive metabolites from the mangrove endophytic fungus Aspergillus sp. GXNU-4QQY1a led to the isolation of two undescribed cyclic peptides, guaspertide A (1) and guaspertide B (2), together with six known compounds, 3-8. These structures and the new compounds' absolute configuration were determined by mass spectrometry analysis, nuclear magnetic resonance spectrum, electronic circular dichroism, and single-crystal X-ray diffraction. Insecticidal assays were carried out with compounds 1-8, and the results showed that compounds 1-3 and 8 exhibited good insecticidal activity against citrus psyllids.
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Inseticidas , Inseticidas/farmacologia , Estrutura Molecular , Aspergillus/química , Fungos , Cristalografia por Raios XRESUMO
Three new compounds (1-2, 14), as well as 22 known compounds (3-13, 15-25), were extracted for the first time from the Selaginella effusa Alston (S. effusa). For the unknown compounds, the planar configurations were determined via NMR and by high-resolution mass spectrometry, while their absolute configurations were determined by calculated electronic circular dichroism (ECD), and the configuration of the stereogenic center of biflavones 4-5 were established for the first time. The pure compounds (1-25) were tested inâ vitro to determine the inhibitory activity of the enzyme-catalyzed reactions. Compounds 1-9 inhibited α-glucosidase with IC50 values ranging from 0.30±0.02 to 4.65±0.04â µM and kinetic analysis of enzyme inhibition indicated that biflavones 1-3 were mixed-type α-glucosidase inhibitors. Compounds 12-13 showed excellent inhibitory activity against urease, with compound 12 (IC50 =4.38±0.31â µM) showing better inhibitory activity than the positive control drug AHA (IC50 13.52±0.61â µM). In addition, molecular docking techniques were used to simulate inhibitor-enzyme binding and to estimate the binding posture of the α-glucosidase and urease catalytic sites.