Tetracyclines are a group of important antibiotics with a common four-ring scaffold. While most tetracyclines are currently used only in animals, their leaching into the environment and residues in food have caused health concerns. Aptamers are an attractive way to detect tetracyclines, and all previously reported aptamers for tetracyclines were obtained by immobilizing target molecules. In this work, we selected a few DNA aptamers by immobilizing the DNA library using oxytetracycline as the target. We obtained new aptamers with no overlapping sequences compared to the previously reported ones, and a representative sequence named OTC5 had a dissociation constant of 147 nM measured by isothermal titration calorimetry. Similar binding affinities were also observed with tetracycline and doxycycline. Because tetracyclines are fluorescent and their fluorescence intensity was enhanced by binding to the aptamers, a label-free and dye-free fluorescent biosensor was developed with a detection limit of 25 nM oxytetracycline. The sensor was able to detect targets in milk after extraction. Fluorescence polarization measurement showed that this aptamer is insensitive to sodium concentration but requires magnesium. Finally, a strand-displacement biosensor was designed, and it has a detection limit of 1.2 μM oxytetracycline.
A simplified predictive model with similar accuracy to a more complex model for predicting independent walking was created, which improves utility in a clinical setting. Such models will allow clinicians to better predict the prognosis of ambulation in individuals who have sustained a traumatic SCI.
Curcumae Rhizoma, known as Ezhu (Chinese: 莪术), and Curcumae Radix, known as Yujin (Chinese: 郁金), are different plant parts coming from three same species according to China Pharmacopoeia. Actually, they are used in different ways in TCM clinical treatment. Curcumae Rhizoma is mainly used as antitumor drug, while Curcumae Radix has been used as antidepressant and cholagogue. Curcumae Rhizoma and Curcumae Radix are confused in variety and source, even in clinical trials by some nonprofessional workers. So it is important for us to make them clear. This review is aimed at summarizing the ethnopharmacology, phytochemical, and pharmacological differences between Curcumae Radix and Curcumae Rhizoma by SciFinder, CNKI, and so on, to use them exactly and clearly. Further studies on Curcumae Rhizoma and Curcumae Radix can lead to the development of new drugs and therapeutics for various diseases on the basis of the TCM theory.
We report a detailed computational and experimental study of the interaction of
single-walled carbon nanotubes (SWCNTs) with the drug-metabolizing cytochrome P450
enzyme, CYP3A4. Dose-dependent inhibition of CYP3A4-mediated conversion of the model
compound, testosterone, to its major metabolite, 6β-hydroxy testosterone
was noted. Evidence for a direct interaction between SWCNTs and CYP3A4 was also
provided. The inhibition of enzyme activity was alleviated when SWCNTs were
pre-coated with bovine serum albumin. Furthermore, covalent functionalization of
SWCNTs with polyethylene glycol (PEG) chains mitigated the inhibition of CYP3A4
enzymatic activity. Molecular dynamics simulations suggested that inhibition of the
catalytic activity of CYP3A4 is mainly due to blocking of the exit channel for
substrates/products through a complex binding mechanism. This work suggests that
SWCNTs could interfere with metabolism of drugs and other xenobiotics and provides a
molecular mechanism for this toxicity. Our study also suggests means to reduce this
toxicity, eg., by surface modification.
Synthesis of colloidal nanocrystals of II-VI semiconductor materials has been refined in recent decades and their size dependent optoelectronic properties have been well established. Here we report a facile synthesis of CdSe-CdS core-shell heterostructures using a two-step hot injection process. Red-shifts in absorption and photoluminescence spectra show that the obtained quantum dots have quasi-type-II alignment of energy levels. The obtained nanocrystals have a heterostructure with a large and highly faceted tetrahedral CdS shell grown epitaxially over a spherical CdSe core. The obtained morphology as well as high resolution electron microscopy confirms that the tetrahedral shell have a zinc blende crystal structure. A phenomenological mechanism for the growth and morphology of the nanocrystals is discussed.
Long noncoding RNAs (lncRNAs) are emerging as important regulators of multiple cellular processes such as cell invasion, growth, apoptosis and differentiation. LncRNAs can function as competing endogenous RNAs (ceRNAs) which sponge and sequester microRNA (miRNA) to regulate specific targets. Previously, we found that the target genes of several miRNAs, including FADD, Fas, Casp and Bax, are related to neuronal apoptosis and form a regulatory network. Among several factors, microRNA‐296‐5p expression was found to be negatively correlated with caspase activity and apoptosis. Here, we aimed to investigate the role of miR‐296‐5p in neuroblastoma (NB) cells. By performing quantitative real‐time PCR (qRT‐PCR), western blot and flow cytometry assays we analysed the expression of apoptotic markers in NB cells transfected with miR‐296‐5p mimics or inhibitor. Pathway‐specific PCR array allowed us to identify the target genes of miR‐296‐5p. Using LncBase online tool, we predicted lncRNA KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1) as an upstream regulator of miR‐296‐5p. The binding of KCNQ1OT1 and miR‐296‐5p was validated via RNA immunoprecipitation and Biotin pull‐down assays. We also demonstrate that miR‐296‐5p suppresses apoptosis of NB cells in vitro and in vivo. Mechanistically, miR‐296‐5p directly bound the 3′UTR of Bax mRNA, thus repressing Bax at the mRNA and protein level. Moreover, through bioinformatic analysis and molecular experiments, we showed that KCNQ1OT1 sponged miR‐296‐5p and impaired its effect on NB cell apoptosis. In summary, KCNQ1OT1 is a potent promoting factor of cell apoptosis, which acts by sponging miR‐296‐5p and upregulating Bax. Our findings identify a regulatory axis of cell fate in NB cells.
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