In vivo cleavage of a target RNA by copper kanamycin A. Direct observation by a fluorescence assay

A novel fluorescence assay to monitor in vivo cleavage chemistry of RNA target sequences has been established and used to demonstrate the activity of copper aminoglycoside mediated degradation of RNA in bacterial cells.     

High‐throughput ultra‐high‐performance liquid chromatography/tandem mass spectrometry quantitation of insulin‐like growth factor‐I and leucine‐rich α‐2‐glycoprotein in serum as biomarkers of recombinant human growth hormone administration

Insulin‐like growth factor‐I (IGF‐I) is a known biomarker of recombinant human growth hormone (rhGH) abuse, and is also used clinically to confirm acromegaly. The protein leucine‐rich α‐2‐glycoprotein (LRG) was recently identified as a putative biomarker of rhGH administration. The combination of an ACN depletion method and a 5‐min ultra‐high‐performance liquid chromatography/tandem mass spectrometry (uHPLC/MS/MS)‐based selected reaction monitoring (SRM) assay detected both IGF‐I and LRG at endogenous concentrations. Four eight‐point standard addition curves of IGF‐I (16–2000 ng/mL) demonstrated good linearity (r² = 0.9991 and coefficients of variance (CVs) <13%). Serum samples from two rhGH administrations were extracted and their uHPLC/MS/MS‐derived IGF‐I concentrations correlated well against immunochemistry‐derived values. Combining IGF‐I and LRG data improved the separation of treated and placebo states compared with IGF‐I alone, further strengthening the hypothesis that LRG is a biomarker of rhGH administration. Artificial neural networks (ANNs) analysis of the LRG and IGF‐I data demonstrated an improved model over that developed using IGF‐I alone, with a predictive accuracy of 97%, specificity of 96% and sensitivity of 100%. Receiver operator characteristic (ROC) analysis gave an AUC value of 0.98. This study demonstrates the first large scale and high throughput uHPLC/MS/MS‐based quantitation of a medium abundance protein (IGF‐I) in human serum. Furthermore, the data we have presented for the quantitative analysis of IGF‐I suggest that, in this case, monitoring a single SRM transition to a trypsin peptide surrogate is a valid approach to protein quantitation by LC/MS/MS. Copyright © 2009 John Wiley & Sons, Ltd.     

Doppler broadening of in-flight positron annihilation radiation due to electron momentum.

We report the first observation of electron momentum contributions to the Doppler broadening of radiation produced by in-flight two-photon annihilation in solids. In these experiments an approximately 2.5 MeV positron beam impinged on thin polyethylene, aluminum, and gold targets. Since energetic positrons easily penetrate the nuclear Coulomb potential and do not cause a strong charge polarization, the experimental annihilation line shapes agree well with calculations based on a simple independent-particle model. Moreover, annihilations with the deepest core electrons are greatly enhanced.     

Tuning the local chemical environment of ZnSe quantum dots with dithiols towards photocatalytic CO2 reduction

Sunlight-driven CO₂ reduction to renewable fuels is a promising strategy towards a closed carbon cycle in a circular economy. For that purpose, colloidal quantum dots (QDs) have emerged as a versatile light absorber platform that offers many possibilities for surface modification strategies. Considerable attention has been focused on tailoring the local chemical environment of the catalytic site for CO₂ reduction with chemical functionalities ranging from amino acids to amines, imidazolium, pyridines, and others. Here we show that dithiols, a class of organic compounds previously unexplored in the context of CO₂ reduction, can enhance photocatalytic CO₂ reduction on ZnSe QDs. A short dithiol (1,2-ethanedithiol) activates the QD surface for CO₂ reduction accompanied by a suppression of the competing H₂ evolution reaction. In contrast, in the presence of an immobilized Ni(cyclam) co-catalyst, a longer dithiol (1,6-hexanedithiol) accelerates CO₂ reduction. ¹H-NMR spectroscopy studies of the dithiol-QD surface interactions reveal a strong affinity of the dithiols for the QD surface accompanied by a solvation sphere governed by hydrophobic interactions. Control experiments with a series of dithiol analogues (monothiol, mercaptoalcohol) render the hydrophobic chemical environment unlikely as the sole contribution of the enhancement of CO₂ reduction. Density functional theory (DFT) calculations provide a framework to rationalize the observed dithiol length dependent activity through the analysis of the non-covalent interactions between the dangling thiol moiety and the CO₂ reduction intermediates at the catalytic site. This work therefore introduces dithiol capping ligands as a straightforward means to enhance CO₂ reduction catalysis on both bare and co-catalyst modified QDs by engineering the particle''s chemical environment.

ZnSe quantum dots (yellow sphere) are modified with dithiols of various lengths for enhanced visible light-driven CO₂ to CO reduction in either the absence or presence of a molecular Ni co-catalyst.