A Water-Soluble Pybox Derivative and Its Highly Luminescent Lanthanide Ion Complexes

A new water-soluble Pybox ligand, <b>1</b>, has been synthesized and found to crystallize in the monoclinic <i>P</i>2₁/<i>n</i> space group with unit cell parameters <i>a</i> = 6.0936(1) Å, <i>b</i> = 20.5265(4) Å, <i>c</i> = 12.0548(2) Å, and β = 90.614(1)°. In the crystal, a water molecule is bound through hydrogen-bonding interactions to the nitrogen atoms of the oxazoline rings. This ligand was used to complex a variety of lanthanide ions, opening up new avenues for luminescence and catalysis in aqueous environment. These complexes are highly luminescent in aqueous solutions, in acetonitrile, and in the solid state. Aqueous quantum yields are high at 30.4% for Eu­(III), 26.4% for Tb­(III), 0.32% for Yb­(III), and 0.11% for Nd­(III). Er­(III) did not luminesce in water, but an emission efficiency of 0.20% could be measured in D₂O. Aqueous emission lifetimes were also determined for the visible emitting lanthanide ions and are 1.61 ms for Eu­(III) and 1.78 ms for Tb­(III). Comparing emission lifetimes in deuterated and nondeuterated water indicates that no water molecules are coordinated to the metal ion. Speciation studies show that three species form successively in solution and the log β values are 5.3, 9.6, and 13.8 for Eu­(III) and 5.3, 9.2, and 12.7 for Tb­(III) for 1:1, 2:1, and 3:1 ligand to metal ratios, respectively

A Water-Soluble Pybox Derivative and Its Highly Luminescent Lanthanide Ion Complexes

A new water-soluble Pybox ligand, <b>1</b>, has been synthesized and found to crystallize in the monoclinic <i>P</i>2₁/<i>n</i> space group with unit cell parameters <i>a</i> = 6.0936(1) Å, <i>b</i> = 20.5265(4) Å, <i>c</i> = 12.0548(2) Å, and β = 90.614(1)°. In the crystal, a water molecule is bound through hydrogen-bonding interactions to the nitrogen atoms of the oxazoline rings. This ligand was used to complex a variety of lanthanide ions, opening up new avenues for luminescence and catalysis in aqueous environment. These complexes are highly luminescent in aqueous solutions, in acetonitrile, and in the solid state. Aqueous quantum yields are high at 30.4% for Eu­(III), 26.4% for Tb­(III), 0.32% for Yb­(III), and 0.11% for Nd­(III). Er­(III) did not luminesce in water, but an emission efficiency of 0.20% could be measured in D₂O. Aqueous emission lifetimes were also determined for the visible emitting lanthanide ions and are 1.61 ms for Eu­(III) and 1.78 ms for Tb­(III). Comparing emission lifetimes in deuterated and nondeuterated water indicates that no water molecules are coordinated to the metal ion. Speciation studies show that three species form successively in solution and the log β values are 5.3, 9.6, and 13.8 for Eu­(III) and 5.3, 9.2, and 12.7 for Tb­(III) for 1:1, 2:1, and 3:1 ligand to metal ratios, respectively

2DG inhibits KIT-glycosylation and KIT-signaling.

<p><b>A</b>. Dose response experiments, after 24h of treatment, examining KIT-signaling in GIST cell lines growing either in HG-media (upper panel), or LG-media (lower panel). <b>B</b>. Time course experiment examining the kinetics of 2DG mediated effects in GIST-T1. <b>C</b>. Effects of co-treatment of GIST-T1 cell with 2DG and mannose on KIT-glycosylation. <b>D</b>. FACS measurement of the expression of KIT on the cell surface of GIST-T1 cells after 2DG and mannose treatment.</p

2DG mediated induction of ER-stress and the UPR A. Dose-response experiments with 2DG (0.01–10mM) show induction of markers of ER-stress and the UPR. B. Inhibition of KIT-glycosylation by tunicamycin, 2DG and withdrawal compared to inhibition of KIT-phosphorylation by IM on the induction of the UPR. C. Effects of withdrawal of Glucose and serum on GIST-T1treated with combinations of mannose and glucose (each 5mM) and 2DG. D. Rescue experiment combining 2DG with mannose and pyruvate (each 5mM).

<p>2DG mediated induction of ER-stress and the UPR A. Dose-response experiments with 2DG (0.01–10mM) show induction of markers of ER-stress and the UPR. B. Inhibition of KIT-glycosylation by tunicamycin, 2DG and withdrawal compared to inhibition of KIT-phosphorylation by IM on the induction of the UPR. C. Effects of withdrawal of Glucose and serum on GIST-T1treated with combinations of mannose and glucose (each 5mM) and 2DG. D. Rescue experiment combining 2DG with mannose and pyruvate (each 5mM).</p

Effects of 2DG in GIST cell lines.

<p><b>A</b>. Cytotoxicity assays (6 days) in IM-sensitive and IM-resistant GIST cell lines with increasing doses of 2DG (50μM—10mM). Dashed line represents GIST-T1 in DMEM-LG media. <b>B</b>. IC50 values depicted as bar chart with dashed line as C_max (0.7mM) from clinical studies (see Refs. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120531#pone.0120531.ref019" target="_blank">19</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120531#pone.0120531.ref020" target="_blank">20</a>). <b>C</b>. Cell cycle analyses after 24h treatment with increasing doses of 2DG. HG: high glucose = 4.6g/L; LG: low glucose = 1.1g/L.</p

Drug combinations of 2DG with IM and ABT-263

<p><b>A</b>. Cytotoxicity assays (3days) combining 2DG with IM and ABT-263. <b>B</b>. Western Blot studies after 24h of co-treatment of 2DG with either IM or ABT-263. <b>C</b>. Induction of apoptosis (caspase 3/7 activation) after 16h of treatment with combinations of 2DG and ABT-263. <b>D</b>. Expression of proapoptotic (BIM) and antiapoptotic (BCL-2, BCL-XL, MCL-1) BCL-2 proteins after single and combined treatment (16h) with 2DG and the BCL-2 antagonist ABT-263.<b>E</b>. Time course studies of 2DG with IM co-treatment (left panel) and 4h IM pre-treatment before addition of 2DG (right panel).</p

Computational discovery of plasma microRNA profiles as biomarkers of temporal lobe epilepsy

<div>Presented at ISMB/ECCB 2015 - July 10 – 14, 2015, Dublin, Ireland</div><div><br></div><div><br></div>Epilepsy is a common neurological disorder affecting approximately 1% of the population and is characterised by recurrent unprovoked seizures. The lack of a clinically accepted biomarker for epilepsy diagnosis as well as the incomplete and vague history often provided by the patient is responsible for up to 30% misdiagnosis. MicroRNAs are a class of small non-coding RNA that regulate gene expression at a post-transcriptional level. MicroRNAs are important contributors to brain function and emerging animal and human data suggest microRNAs control multiple pathways in epilepsy. MicroRNAs are also detectable in various body fluids and their stability as well as link to disease mechanism makes them potentially ideal molecular biomarkers of epilepsy. We determined plasma levels of over 800 microRNAs collected from 20 healthy volunteers and 20 epilepsy patients using highthroughput real-time quantitative reverse transcription PCR. Computational analysis included normalisation, clustering, differential expression analysis, target prediction and pathway analysis. A number of significantly differentially expressed microRNAs were identified between control and epilepsy samples including known brain-expressed microRNAs implicated in epilepsy. Furthermore, we applied feature selection with machine learning algorithms, including support vector machines and bidirectional recurrent neural networks, to build a microRNAs-based predictor of epilepsy, validated on an independent test set. This analyse showed that these classifiers may be useful in supporting the existence of a set of microRNAs implicated in disease pathogenesis that may be biomarkers of human epilepsy

Indazole-Based Covalent Inhibitors To Target Drug-Resistant Epidermal Growth Factor Receptor

The specific targeting of oncogenic mutant epidermal growth factor receptor (EGFR) is a breakthrough in targeted cancer therapy and marks a drastic change in the treatment of non-small cell lung cancer (NSCLC). The recurrent emergence of resistance to these targeted drugs requires the development of novel chemical entities that efficiently inhibit drug-resistant EGFR. Herein, we report the optimization process for a hit compound that has emerged from a phenotypic screen resulting in indazole-based compounds. These inhibitors are conformationally less flexible, target gatekeeper mutated drug-resistant EGFR-L858R/T790M, and covalently alkylate Cys797. Western blot analysis, as well as characterization of the binding kinetics and kinase selectivity profiling, substantiates our approach of targeting drug-resistant EGFR-L858R/T790M with inhibitors incorporating the indazole as hinge binder

<i>TP53</i> sequencing results of 62 patients with primary localized (n = 40) or metastastic (n = 22) GIST.

<p><i>TP53</i> sequencing results of 62 patients with primary localized (n = 40) or metastastic (n = 22) GIST.</p