Dega, Naresh KumarGanganboina, Akhilesh BabuTran, Hai LinhKuncoro, Eko Prasetyo...
10页
查看更多>>摘要：The development of an efficient protein-inorganic nanohybrid with superior nanozyme activity for highly sensitive detection of glutathione (GSH) is essential for early diagnosis of human diseases. Herein, a rapid and highly sensitive colorimetric assay using self-assembled bovine serum albumin-hydrated manganese phosphate nano-flowers (MnPNF) as a biomimic oxidase is developed for GSH detection in human serum. The BSA can complex with Mn2+ to serve the nucleation center to produce MnPNF in the presence of phosphate-buffered saline (PBS). The morphology and surface characterization results show that the MnPNF is assembled with hierarchical nanoplates to form 500 nm nanoflowers. The oxidase-like activity of MnPNF is based on the redox reaction with 3,3',5,5'-tetramethylbenzidine. However, the addition of GSH can reduce MnPNF to Mn2+, and subsequently supresses the oxidase-like activity and a yellow color at 450 nm is observed in the presence of H2SO4. The MnPNF-based nanozyme exhibits excellent sensing ability toward GSH detection, and a good linear relationship between the change in absorbance at 450 nm and the added amounts of GSH at 50 nM-10 mu M with low limits of detection of 20 and 26.6 nM in the PBS and diluted human serum, respectively, is observed. Moreover, the sensing probe shows a superior selectivity over the other 16 interferences, which drive the determination of GSH feasible in real human serum. Since the MnPNF can be simply prepared at room temperature and no functionalization is required, this assay can be used to design the highly efficient biomimic oxidase for effective sensing of GSH and other disease-related biomolecules in biological fluid samples.
查看更多>>摘要：The monitoring of the fungal genus Alternaria, which causes destructive brown spot disease in citruses worldwide and produces highly toxic mycotoxins, is extremely important to protect citrus and human health. In this work, we describe an ultrasensitive colorimetric method for the detection of genomic DNA of Alternaria from citrus fruit samples, using a system consisting of five groups of reporter probes. Each reporter probe is prepared by coupling recognition DNA and horseradish peroxidase (HRP) on the surface of gold nanoparticle (AuNP) through a convenient and low-cost freezing-assisted method. Meanwhile, the capture DNA is immobilized on magnetic bead (MB) via biotin-streptavidin reaction. Then, the capture DNA, target DNA, and five groups of AuNP-based reporter probes form a stable DNA-heptamer sandwich structure on the MB, and then HRP generates a blue signal for the subsequent colorimetric detection. It should be noted that AuNP with a large specific surface area drives abundant HRP anchoring, resulting in significant signal amplification. In addition, there are five groups of AuNPbased reporter probes, which further amplify the detection signal. As a result, the detection limit of the artificial target DNA is as low as 15.6 pM. Because the detection signal can be recorded visually without any special equipment, and its sensitivity is high, this method represents a suitable diagnostic tool for Alternaria genetic detection.
查看更多>>摘要：A rapid colorimetric method for detecting sodium benzoate in food products was established based on the D-amino acid oxidase (DAAO) and 2D metal organic framework (2D MOF) nanosheets mediated cascade enzyme reactions. Firstly, the synthesized 2D MOF nanosheets served as high efficient nanozyme with outstanding peroxidase-like catalytic activity and catalyzed the color reaction between H2O2 and 3, 3', 5, 5'-tetramethylbenzidine. Secondly, sodium benzoate as a competitive inhibitor of DAAO, could influence the production of H2O2 in DAAO mediated oxidation reaction. After a combination of those two reactions, this colorimetric quantitative method was constructed and validated for sodium benzoate determination with wide linear range (2.0-200.0 mu M), low limit of detection (2.0 mu M), high accuracy (recovery rate in 95.80-108.00%) and satisfied selectivity. Lastly, this method was utilized to analyze sodium benzoate concentration in juice, wine and vinegar by naked eyes.
查看更多>>摘要：Raman spectroscopy combined with artificial intelligence algorithms have been widely explored and focused on in recent years for food safety testing. It is still a challenge to overcome the cumbersome culture process of bacteria and the need for a large number of samples, which hinder qualitative analysis, to obtain a high classification accuracy. In this paper, we propose a method based on Raman spectroscopy combined with generative adversarial network and multiclass support vector machine to classify foodborne pathogenic bacteria. 30,000 iterations of generative adversarial network are trained for three strains of bacteria, generative model G generates data similar to the actual samples, discriminant model D verifies the accuracy of the generated data, and 19 feature variables are obtained by selecting the feature bands according to the Raman spectroscopy pattern. Better classification results are obtained by optimising the parameters of the multi-class support vector machine, etc. Our detection and classification method not only solves the problem of needing a large number of samples as training set, but also improves the accuracy of the classification model. Therefore, this GAN-SVM classification model provides a new idea for the detection of bacteria based on Raman spectroscopy technology combined with artificial intelligence algorithms.
查看更多>>摘要：Sensitivity and credibility detecting histamine (HA) as an important neurotransmitter in biofluids is of importance in analytical science and physiology. Surface-enhanced Raman spectroscopy (SERS) is able to realize the high sensitivity with single molecules level, but providing the high sensitivity for HA with a small cross section remains a challenge. Here we develop the metal complex-based SERS nanoprobe nitrilotriacetic acid-Ni2+ (NTA-Ni2+) combined with self-assemble Au NPs active substrates for sensitive detection of HA. The NTA-Ni2+ can capture the HA molecules close to Au NPs substrates and then amplify the Raman signals of HA owing to the formation of a complex of NTA-Ni2+-HA. The self-assemble Au film through the evaporation-driven method can provide the high-density hot spots substrate with high stability and reproducibility. The NTA-Ni2+ decorated Au NPs as nanoprobe responds to HA with 1 mu M level of sensitivity. More importantly, the developed SERS nanoprobe composing of NTA-Ni2+ and self-assemble Au NPs can be utilized to detect and monitor the HA spiked into serum, indicating the potential prospect in analysis of HA in complex specimen.
查看更多>>摘要：Heterogeneous analysis has great application prospects in the detection of post-translational modification (PTM) enzymes with the advantages of signal enhancement, less sample demand, and high sensitivity and selectivity. Nevertheless, once the substrate was fixed on a solid interface, the steric hindrance might limit the approaching of catalytic center to the substrate, thus reducing the efficiency of PTM. Herein, we suggested that the avidinmodified interface could be used to develop heterogeneous sensing platforms with biotin-labeled substrates as the probes, in which the enzymatic PTM was performed in solution and the heterogeneous assay was conducted on a solid surface. The sensing strategy integrates the advantages but overcomes the defects of both homogeneous and heterogeneous assays. Protein kinase A (PKA) and histone acetyltransferase (HAT) were determined as the examples by using sequence-specific peptide substrates. The signal changes were monitored by HRP-based colorimetric assay and antibody-amplified surface plasmon resonance (SPR). The methods were used for analysis of cell lysates and evaluation of inhibition efficiency with satisfactory results. The strategy can be used for the detection of a variety of biological enzymes and provide a new idea for the design of various heterogeneous biosensors. Thus, this work should be of great significance to the popularization and practical application of biosensors.
查看更多>>摘要：A nucleolus as a prominent sub-nuclear, membraneless organelle plays a crucial role in ribosome biogenesis, which is in the major metabolic demand in a proliferating cell, especially in aggressive malignancies. We develop a gamma-glutamyltranspeptidase (GGT)-activatable indole-quinolinium (QI) based cyanine consisting of a novel tripeptide fragment (Pro-Gly-Glu), namely QI-PG-Glu as a turn-on red fluorescent probe for the rapid detection of GGT-overexpressed A549 cancer cells in vivo. QI-PG-Glu can be triggered by GGT to rapidly release an activated fluorophore, namely HQI, in two steps including the cleavage of the gamma-glutamyl group recognized by GGT and the rapid self-driven cyclization of the Pro-Gly linker. HQI exhibits dramatically red fluorescence upon binding to rRNA for imaging of nucleolus in live A549 cells. HQI also intervenes in rRNA biogenesis by declining the RNA Polymerase I transcription, thus resulting in cell apoptosis via a p53 dependent signaling pathway. Our findings may provide an alternative avenue to develop multifunctional cancer cell-specific nucleolus-targeting fluorescent probes with potential anti-cancer effects.
Aparicio-Muriana, M. MarCarmona-Molero, RocioLara, Francisco J.Garcia-Campana, Ana M....
11页
查看更多>>摘要：The presence of cyanobacteria and cyanotoxins in all water bodies, including ocean water and fresh water sources, represents a risk for human health as eutrophication and climate change are enhancing their level of proliferation. For risk assessment and studies on occurrence, the development of reliable and sensitive analytical approaches able to cover a wide range of cyanotoxins is essential. This work describes the development of an HILIC-MS/MS multiclass method for the simultaneous analysis of eight cyanotoxins in reservoir water samples belonging to three different classes according to their chemical structure: cyclic peptides (microcystin-LR, microcystin-RR and nodularin), alkaloids (cylindrospermopsin, anatoxin-a) and three non-protein amino acids isomers such as beta-methylamino-L-alanine, 2,4-diaminobutyric acid and N-(2-aminoethyl)glycine). A SeQuant ZIC-HILIC column was employed to achieve the chromatographic separation in less than 12 min. Previously, a novel sample treatment based on a tandem solid-phase extraction (SPE) system using mixed cation exchange (MCX) and Strata-X cartridges was investigated with the aim of extracting and preconcentrating this chemically diverse group of cyanotoxins. The Strata-X cartridge, which was configured first in the line of sample flow, retained the low polar compounds and the MCX cartridge, which was at the bottom of the dual system, retained mainly the non-protein amino acids. The optimization procedure highlighted the importance of sample ion content for the recoveries of some analytes such as the isomers beta-N-methylamino-L-alanine and 2-4-diaminobutyric acid. Method validation was carried out in terms of linearity, limit of detection (LOD) and quantification (LOQ), recoveries, matrix effect and precision in terms of repeatability and intermediate precision. This work represents the first analytical method for the simultaneous analysis of these multiclass cyanotoxins in reservoir water samples, achieving LOQs in the very low range of 7.10(-3) - 0.1 mu g L-1. Despite high recoveries obtained at the LOQ concentration levels (101.0-70.9%), relative standard deviations lower than 17.5% were achieved.
查看更多>>摘要：Pioglitazone is a Peroxisome Proliferator-Activated Receptor (PPAR) agonist of the thiazolidinedione class of compounds with promising anticancer activity. An innovative quantitative mass spectrometry imaging (MSI) method and a HPLC-UV method were developed and validated to investigate its distribution in tumor and liver tissues. The MSI method is based on stable isotope normalization and resulted highly specific and sensitive (0.2 pmol/spot). The correct identification of the drug ion signal is confirmed by MS/MS analysis on tissue. The method shows an optimal lateral resolution (25 mu m) relying on the ionization efficiency and fine laser diameter of the atmospheric pressure MALDI source. The HPLC-UV method is simple and straightforward involving quick protein precipitation and shows good sensitivity (50ng/sample) using a small starting volume of biological sample. Thus, it is applicable to samples obtained from both preclinical models and clinical surgical procedures. MSI and HPLC-UV assays were validated assessing linearity, intra- and inter-day precision and accuracy, limit of quantification, selectivity and recovery. These are the first methods developed and validated for the analysis of pioglitazone in tissues, and they were applied successfully to myxoid liposarcoma xenograft-bearing mice, which received clinically relevant drug doses. Pioglitazone was measured by either method in sections of tumor and liver 2, 6 and 24 h post-treatment. Drug distribution was relatively homogeneous.
查看更多>>摘要：Endoplasmic reticulum (ER) is an indispensable organelle responsible for protein synthesis, transportation, and maintenance of Ca2+ homeostasis in eukaryotic cells. Recent studies highlighted that ER-targeted photosensitizers with high yield of singlet oxygen (O-1(2)) are effective in selectively disrupting ER function and are promising candidates for anticancer therapy. Unfortunately, no ER targetable fluorescent probes for determining O-1(2) photosensitized in this photodynamic therapy process is available. In this work, we synthesized an ER-targetable, two-photon fluorescence probe, ER-O-1(2), for fluorescence turn-on sensing of O-1(2). ER-O-1(2) demonstrated high sensitivity to O-1(2) sensing with a wide detection range (0-2.75 mu M) and a low detection limit (0.11 mu M). ER-O-1(2) also displayed excellent selectivity toward O-1(2) out of other ROS and metal ions. Notably, ER-O-1(2) exhibited low cytotoxicity but with specific ER targetable capability. On account of these advantageous features, fluctuations of O-1(2) in living cells and brain tissues were effectively visualized by ER-O-1(2).
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Elsevier