Herbertschneider1624

1% deletion abundance. The results were 100% consistent with ARMS-PCR for the 38 tumor tissues tested and were in good agreement with next-generation sequencing for quantifying the abundance of EGFR 19-Del in 15 cfDNA samples, showing the great potential of the method for liquid biopsies.Developing a green analytical method for the analysis of components in food samples is an important research aspect of liquid chromatography (LC). The traditional LC method usually consumes a lot of toxic solvent for sample extraction and LC separation. In the current study, a green analytical method for the rapid determination of ergosterol in edible fungi was established. The sample was extracted and purified by matrix solid-phase dispersion (MSPD) with a green solution (ethanol and water). The LC separation was performed using a Poroshell 120 SB-C18 (4.6 × 30 mm, 2.7 μm) column with a green mobile phase (94% ethanol) at a flow rate of 1.0 mL min-1. The detection wavelength was set at 283 nm. The calibration curve of ergosterol showed good linearity (R = 0.9999) within the test range (4.21-25.27 μg mL-1). The RSD of precision was less than 2.0% and the recovery was 100.4% (RSD = 3.23%). The developed method was successfully applied to quantitative analysis of ergosterol in six edible fungi and the contents of ergosterol were in the range of 1.68-4.02 mg g-1. Only 11.5 mL ethanol water solution was used in the sample extraction and LC separation in the newly developed method, and no toxic organic solvents were used. The total analysis time was less than 15.5 min, about 12-14 min for sample extraction and 1.5 min for LC analysis. This method was environmentally friendly and time-saving, which is helpful to improve the quality evaluation of edible fungi.A novel mitochondrial-targeted deep-red fluorescence ATP probe, NIR-A, is reported. The probe showed a fast, selective, and reversible response for ATP with a significant turn-on fluorescence signal at 663 nm with a large Stokes shift of 81 nm. Additionally, the introduction of TPP enabled TPP-endowed NIR-A to be enriched predominantly in the mitochondria. NIR-A was successfully applied to monitor ATP fluctuation in Ramos cells and zebrafish in real-time with good biocompatibility.Whole blood analysis reveals crucial information about various physiological and pathological conditions, including cancer metastasis, infection, and immune status, among others. Despite this rich information, the complex composition of whole blood usually required multiple sample preparation steps to purify targeted analytes. Traditionally, whole blood preparation processes, including centrifugation, lysis, dilution, or staining, are usually manually operated by well-trained technicians using bench-top instruments. This preparation can require a large blood volume and cannot be directly integrated with detection systems. Recently, various studies have integrated microfluidics with electrical sensors for whole blood analysis, with a focus on cell-based analysis, such as cell type, number, morphology, phenotype, and secreted molecules. These miniaturized systems require less sample and shorter reaction times. Besides, the sample processing and analysis can be fully integrated and automated with minimal operations. We believe these systems can transfer the current whole blood analysis from hospitals or laboratories into clinics or home settings to enable real-time and continuous health condition monitoring in point-of-care settings.A metal-free oxidative cascade acylation and dearomatization of N-(p-methoxyaryl)propiolamides was achieved via K2S2O8 mediated decarboxylation of α-oxocarboxylic acids under operationally simple conditions to access azaspiro[4,5]-trienones in good to excellent yields. Furthermore, the utility of the protocol was illustrated in a one-pot reaction sequence consisting of Ugi-reaction/spirocyclization/aza-Michael transformation for the construction of complex tricyclic cores having quaternary spirocenters.This paper describes a new label-free fluorescent aptasensor for the detection of aflatoxin B1 (AFB1) based upon exonuclease I (Exo I) and SYBR Gold, in which SYBR Gold, aptamer, AFB1, and Exo I were used. Specific combinations of aptamer and AFB1 occurred in the presence of AFB1 and consequently altered the spatial structure of the aptamer, thereby preventing its digestion by Exo I. When SYBR Gold was added, intense fluorescence was observed. Additionally, a good linear relationship was observed under optimized conditions between the fluorescence intensities and the AFB1 concentrations (R2 = 0.993). The established aptamer sensor was highly sensitive and exhibited a low limit of detection of 1.82 ng mL-1, with superior specificity for AFB1. It was also used in the quantification of AFB1 levels in soybean sauce samples and demonstrated satisfactory recoveries in the scope of 94.8-108.9%. The proposed sensor is highly sensitive, low cost, and capable of rapid detection and can thus be used to determine mycotoxin levels in a wide range of feeds and food products in a high-throughput and quantitative means.A universal method to measure the binding affinities of antibody drugs towards their targets on the surface of living cells was developed based on atomic force microscopy (AFM) analysis. Nivolumab, an antibody drug targeting programmed cell death 1 (PD-1), was mainly used as a model for this evaluation. The surface of a tip-less AFM cantilever was coated with nano-capsules, on which immunoglobulin G-binding ZZ domains of protein A were exposed, and nivolumab molecules were immobilized on the cantilever through binding between the antibody Fc domains and the ZZ domains, which controlled the molecular orientation of the antibodies. Model human T lymphocytes (Jurkat), on which PD-1 molecules were highly expressed, were immobilized on a glass substrate via a lipid bilayer-anchoring reagent. The nivolumab-coated AFM cantilever was moved to approach the T cells, and the rupture forces between nivolumab molecules on the AFM cantilever and PD-1 molecules on the cell surface were measured. The average values of the ruingle cell. These results indicated that the present method is useful for selecting the most effective candidates from various antibody drugs from the point of view of binding forces between antibodies and living cells.Herein, we have reported a new one-step potentiometric immunoassay for the sensitive and specific detection of human plasma cardiac troponin I (cTnI), a biomarker of cardio-cerebrovascular diseases. Initially, the cTnI biomolecules were immobilized on the surface of a gold nanoparticle-functionalized screen-printed graphite electrode (SPGE). Thereafter, rabbit polyclonal antibodies to cTnI were covalently conjugated to the bis-MPA-COOH dendrimers through typical carbodiimide coupling. The introduction of the target analyte caused a competitive immunoreaction between the immobilized cTnI on the electrode and the conjugated antibody on the dendrimers. The potentiometric measurement was mainly derived from the change in the surface charge on the surface of the modified electrode due to the negatively charged bis-MPA-COOH dendrimers after the immunoreaction. On increasing target cTcI, the number of charged dendrimers on the immunosensor decreased, resulting in a change in the electric potential. Under optimum conditions, the potentiometric immunosensor exhibited good potentiometric responses for the detection of cTcI and allowed the determination of the target analyte at a concentration as low as 7.3 pg mL-1. An intermediate precision of ≤8.7% was accomplished with batch-to-batch identification. Meanwhile, the potentiometric immunosensor showed good anti-interfering capacity and selectivity against other proteins and biomarkers. Importantly, our system displayed high accuracy for the analysis of human plasma serum samples containing target cTcI relative to commercial human cTcI enzyme-linked immunosorbent assay (ELISA) kits.In the pharmaceutical industry, finding cost-effective and real-time analyzers that provide valid data is a good aim. The purpose of this work was to propose a link between the pharmaceutical industry and the recent innovations in solid-contact ion-selective electrodes (SC-ISEs) for the utilization of these electrodes as real-time analyzers to evaluate the concentration of tetrahydrozoline HCl in different matrices. The backbone of these new potentiometric sensors is the conjunction of calix[6]arene and (2-hydroxypropyl)-β-cyclodextrin as molecular recognition elements and a network of multi-walled carbon nanotubes as a solid transducer material between an ionophore-doped PVC membrane and microfabricated Cu electrodes. The proposed sensors were optimized to determine tetrahydrozoline, and their performances were assessed according to the IUPAC recommendations. The proposed solid-contact sensors were compared with liquid contact sensors, and the former sensors were found to be better than the latter sensors in terms of durability, handling, and easier adaptation to industry with comparable sensitivity. The measurements were implemented using phosphate buffer (pH 6). The best obtained linearity range was 1 × 10-2 to 1 × 10-7 M, and the best LOD was 1 × 10-8 M. The sensors with the best performance were successfully applied to determine tetrahydrozoline in a pharmaceutical eye preparation and rabbit tears. The obtained results were statistically compared to those obtained by the official method of analysis, and no significant difference was obtained. The eco-score of the method was assessed using the eco-scale tool and also compared with that of the official method. The proposed approach was validated according to the International Council for Harmonisation (ICH) guidelines.This study proposes a new multi-residue enantioselective method for the determination of emerging drug contaminants in sea water by solid phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). To achieve satisfactory enantiomeric separation with a vancomycin stationary phase it was essential to limit sodium chloride in extracted samples to less then 1 μg per injection. This was achieved through a straightforward SPE method using a 50 mL water wash volume and analyte elution in acetonitrile. A Chiral-V enantioselective column (150 × 2.1 mm; 2.7 μm particle size) operated in polar ionic mode enabled simultaneous drug separations in 30 minutes. Analytes with enantioresolution ≥1 were the stimulants amphetamine and methamphetamine, the beta-agonist salbutamol, the beta-blockers propranolol, sotalol and acebutolol, the anti-depressants fluoxetine, venlafaxine, desmethylvenlafaxine and citalopram, and the antihistamine chlorpheniramine. Method quantitation limits were less then 10 ng L-1 and method trueness was 80-110% for most analytes. The method was applied to samples from the Forth and Clyde estuaries, Scotland. Chiral drugs were present at concentrations in the range 4-159 ng L-1 and several were in non-racemic form (enantiomeric fraction ≠ 0.50) demonstrating enantiomer enrichment. This emphasises the need for further enantiospecific drug exposure and effect studies in the marine environment.