Label-free and sensitive detection of RNA demethylase FTO with primer generation rolling circle amplification

We develop for the first time a label-free fluorescent method for sensitive detection of fat mass and obesity-associated protein (FTO) activity using MazF-mediated primer generation rolling circle amplification. This method...

Supramolecular complexation between cucurbit[7]uril and folate and analytical applications

Folate (FA) plays a key role in the biosynthesis of amino acids, purines, and pyrimidines in the human body, and intracellular folate metabolism has become an attractive target of tumor chemotherapy. In this work, an inclusion interaction was found between FA and cucurbit[7]uril (CB[7]), and the formation of a CB[7]-FA 2:1 supramolecular inclusion complex was confirmed by fluorescence spectra, UV-Vis absorption spectroscopy, 1H NMR, and molecular modeling calculations. In addition, FA is generally determined through the indirect fluorescent method because it shows weak fluorescence in aqueous solution. Therefore, a simple, direct fluorescence probe method for rapidly measuring FA was investigated, and the linear equation of FA was ΔF = 14.691C + 37.366 within the concentration ranges of 0.82 ~ 18.31 µg mL–1. The proposed direct fluorescence method was applied to the determination of spiked plasma. We demonstrated that this method could provide an experimental basis for the targeted administration of the CB[7]-FA complex, and it could be extended as a promising fluorescence detection method for drugs in vivo.

Synthesis of Symmetrical Acetophenone Azine Derivatives as Colorimetric and Fluorescent Cyanide Chemosensors

Cyanide is a highly toxic anion and poison to the environment. Therefore, fast, effective, and efficient analysis methods to detect cyanide are needed. Herein, symmetrical chemosensor of 2’-hydroxy acetophenone azine (1) and 2’,4’-dihydroxy acetophenone azine (2) has been synthesized tested as colorimetric and fluorescent cyanide chemosensor. The azines were produced from the condensation of acetophenone derivatives with hydrazine hydrate in ethanol under reflux or ultrasonic irradiation methods. Colorimetric and fluorescent chemosensor tests showed selectivity to acetate and cyanide anions in DMSO. The limit of detection (LOD) for colorimetric measurement of cyanide anion was 9.68×10–4 M for compound (1) and 9.63×10–5 M for compound (2), while the fluorescent method showed 15.90×10–4 M for compound (1) and 8.95×10–5 M for compound (2), respectively. In addition, test paper-strips containing sensor 2 indicated noticeable results for 'naked eye' detection of cyanide in an aqueous medium.

Electrochemical and Fluorescent Properties of Crown Ether Functionalized Graphene Quantum Dots for Potassium and Sodium Ions Detection

The concentration of sodium and potassium ions in biological fluids, such as blood, urine and sweat, is indicative of several basic body function conditions. Therefore, the development of simple methods able to detect these alkaline ions is of outmost importance. In this study, we explored the electrochemical and optical properties of graphene quantum dots (GQDs) combined with the selective chelating ability of the crown ethers 15-crown-5 and 18-crown-6, with the final aim to propose novel composites for the effective detection of these ions. The results obtained comparing the performances of the single GQDs and crown ethers with those of the GQDs-15-crown-5 and GQDs-18-crown-6 composites, have demonstrated the superior properties of these latter. Electrochemical investigation showed that the GQDs based composites can be exploited for the potentiometric detection of Na+ and K+ ions, but selectivity still remains a concern. The nanocomposites showed the characteristic fluorescence emissions of GQDs and crown ethers. The GQDs-18-crown-6 composite exhibited ratiometric fluorescence emission behavior with the variation of K+ concentration, demonstrating its promising properties for the development of a selective fluorescent method for potassium determination.

The Proteasome Inhibitor Bortezomib Induces Apoptosis and Activation in Gel-Filtered Human Platelets

Bortezomib (BTZ) has demonstrated its efficacy in several hematological disorders and has been associated with thrombocytopenia. There is controversy about the effect of BTZ on human platelets, so we set out to determine its effect on various types of platelet samples. Human platelets were investigated in platelet-rich plasma (PRP) and as gel-filtered platelets (GFPs). Mitochondrial inner membrane potential depolarization and phosphatidylserine (PS) and P-selectin expression levels were studied by flow cytometry, while thrombin generation was measured by a fluorescent method. In PRP, BTZ caused negligible PS expression after 60 min of treatment. However, in GFPs, PS expression was dose- and time-dependently increased in the BTZ-treated groups, as was P-selectin. The percentage of depolarized cells was also higher after BTZ pretreatment at both time points. Peak thrombin and velocity index increased significantly even with the lowest BTZ concentration (p = 0.0019; p = 0.0032) whereas time to peak and start tail parameters decreased (p = 0.0007; p = 0.0034). The difference between PRP and GFP results can be attributed to the presence of plasma proteins in PRP, as the PS-stimulating effect of BTZ could be attenuated by supplementing GFPs with purified human albumin. Overall, BTZ induces a procoagulant platelet phenotype in an experimental setting devoid of plasma proteins.

Highly Sensitive and Selective Fluorescence “Turn-On” Detection of Pb (II) Based on Fe3O4@Au–FITC Nanocomposite

New nanocomposites, Fe3O4@Au–FITC, were prepared and explored to develop a fluorescent detection of Pb2+. The Fe3O4@AuNPs–FITC nanocomposites could be etched by Pb2+ in the presence of Na2S2O3, leading to fluorescence recovery of FITC quenched by Fe3O4@Au nanocomposites. With the increase of Pb2+ concentration, the fluorescence recovery of Fe3O4@AuNPs–FITC increased gradually. Under optimized conditions, a detection limit of 5.2 nmol/L of Pb2+ with a linear range of 0.02–2.0 µmol/L were obtained. The assay demonstrated negligible response to common metal ions. Recoveries of 98.2–106.4% were obtained when this fluorescent method was applied in detecting Pb2+ spiked in a lake-water sample. The above results demonstrated the high potential of ion-induced nanomaterial etching in developing robust fluorescent assays.

A Fluorescent Detection for Paraquat Based on β-CDs-Enhanced Fluorescent Gold Nanoclusters

In this report, a fluorescent sensing method for paraquat based on gold nanoclusters (AuNCs) is proposed. It was found that paraquat could quench both glutathione-capped AuNCs (GSH-AuNCs) and β-cyclodextrin-modified GSH-AuNCs (GSH/β-CDs-AuNCs). The modification of β-CDs on the surface of GSH-AuNCs obviously enhanced the fluorescence intensity of GSH-AuNCs and improved the sensitivity of paraquat sensing more than 4-fold. This sensibilization was ascribed to the obvious fluorescence intensity enhancement of GSH-AuNCs by β-CDs and the “host–guest” interaction between paraquat and β-CDs. The fluorescence quenching was mainly due to the photoinduced energy transfer (PET) between GSH/β-CDs-AuNCs and paraquat. With the optimized β-CDs modification of the GSH-AuNC surfaces and under buffer conditions, the fluorescent detection for paraquat demonstrated a linear response in the range of 5.0–350 ng/mL with a detection limit of 1.2 ng/mL. The fluorescent method also showed high selectivity toward common pesticides. The interference from metal ions could be easily masked by ethylene diamine tetraacetic acid (EDTA). This method was applied to the measurement of paraquat-spiked water samples and good recoveries (93.6–103.8%) were obtained. The above results indicate that host molecule modification of fluorescent metal NC surfaces has high potential in the development of robust fluorescent sensors.

SEASONAL AND INTERANNUAL DYNAMICS OF CHLOROPHYLL IN PLANKTON OF THE RYBINSK RESERVOIR (2015–2019)

New data on chlorophyll content at standard stations in the Rybinsk reservoir, that continue series of long-term observations, were obtained in May–October 2015–2019. A fluorescent method is used in this study which makes it possible to determine the total amount of chlorophyll (ΣChl) by its content in cyanoprokaryotes, di-atoms, and green algae (ChlCyan, ChlBac, ChlChl, respectively). These taxa are the main contributors to ƩChl which show interannual and seasonal variation. In different years, the average content of ChlBac makes 2.4–6.6 μg/L or 24–50% of the ΣChl pool, ChlCyan – 3.1–10.9 μg/L (50–70%), ChlChl – 0.3–1.2 μg/L (3–12%). The distribution of ChlBac in the water column is generally uniform while the amount of ChlCyan decreases with depth. With the average ΣChl content for the growing season 14.8±1.0, 13.0±1.3, 7.8±0.6, 13.6±1.6, and 11.9±1.0 μg/L in 2015–2019, the trophic status of the reservoir varied from moderately eutrophic to mesotrophic.