The formation of a fluorescent photoadduct between 5-fluoro-4-thiouridine ((FS) U), in the sequence context 5'-A(FS) UA-3' and incorporated into a synthetic oligonucleotide either at its 3'- or 5'-end, and one of the thymines of the TAT motif in a complementary target DNA strand led to photo-crosslinking of the two strands for several oligonucleotide constructs. Enzymatic digestion, MS, UV, and fluorescence spectral analyses of the interstrand crosslinked oligonucleotides revealed the identity of the thymidine that participates in the photo-crosslinking reaction as well as the diastereomeric structures of the crosslinks. The proposed pathways of interstrand photo-crosslinking are supported by experiments with isotopically labeled oligonucleotide constructs and visualized by means of molecular dynamics simulations.
The antioxidant potential of bird cherry fruit of water, methanol, ethanol, and acetone extracts and their antioxidant efficiency against oxidation of PC liposomes using spectroscopic and chromatographic methods were investigated. The chromatographic methods quantified and specified the presence of phenolic and flavonoid compounds in the investigated extracts. The characteristic peaks in the UV spectrum at 275 nm and 370 nm confirmed the presence of phenols and flavonoids and their derivatives. Their presence was also confirmed by FTIR spectra, which revealed the presence of its functional groups. The total luminescence spectra with maxima at 314–318 nm, 325–355 nm, and 428–435 nm were ascribed to the presence of phenolic acids and tocopherols. The antioxidant properties of extracts and its inhibition properties against lipid peroxidation in PC liposomes were determined by fluorogenic probes DCF-H and C11-BODIPY581/591. The measured antioxidant properties against generated free radicals in aqueous and lipogenic phases revealed differences between extracts depending on their physicochemical properties with the greatest potential for acetone extract and sirup. The presented quantitative analysis indicated that cherry bird extracts possess significant amounts of phenolics and flavonoids, thus having the opportunity to be used as a natural antioxidant agent source with a large potential for application in pharmaceutical and food industries.
The interactions of G–quadruplexes of different topologies with highly fluorescent 9-methoxyluminarine ligand 9-MeLM were investigated by fluorescence and circular dichroism spectroscopy. The results showed that 9-methoxyluminarine was able to interact and did not destabilize any investigated molecular targets. The studied compound was selectively quenched by parallel c-MYC G-quadruplex DNA, whereas hybrid and antiparallel G4 topology caused only a negligible decrease in the fluorescence of the ligand. A high decrease of fluorescence of the ligand after binding with c-MYC G-quadruplex suggests that this molecule can be used as a selective probe for parallel G-quadruplexes.
The interstrand photocrosslinking of a 5-chloro-2′-deoxy-4-thiouridine modified DNA oligonucleotide proceeds via a two-step photocascade involving the formation of a thermally reversible crosslink and its conversion to a stable fluorescent crosslink.
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