X-Ray Crystallography and Free Energy Calculations Reveal the Binding Mechanism of A2A Adenosine Receptor Antagonists

We present a robust protocol based on iterations of free energy perturbation (FEP) calculations, chemical synthesis, biophysical mapping and X-ray crystallography to reveal the binding mode of an antagonist series to the A_2A adenosine receptor (AR). Eight A_2AAR binding site mutations from biophysical mapping experiments were initially analyzed with sidechain FEP simulations, performed on alternate binding modes. The results distinctively supported one binding mode, which was subsequently used to design new chromone derivatives. Their affinities for the A_2AAR were experimentally determined and investigated through a cycle of ligand-FEP calculations, validating the binding orientation of the different chemical substituents proposed. Subsequent X-ray crystallography of the A_2AAR with a low and a high affinity chromone derivative confirmed the predicted binding orientation. The new molecules and structures here reported were driven by free energy calculations, and provide new insights on antagonist binding to the A_2AAR, an emerging target in immuno-oncology.     

Micrometer scale gel patterns

A novel method for fabricating micrometer sized gel patterns is described. The presented method involves spin-coating a pre-gel solution on a surface that was chemically treated to modulate its surface energy, creating highly hydrophobic areas on a hydrophilic substrate. Following spin-coating, the gel solution self organizes on the hydrophilic sites. This method offers the advantages of high resolution, self-alignment to pre-patterned electrodes, and a simple straightforward fabrication process. Minimum feature size achieved was approximately 20 μm. The characteristic shrinking and swelling times of gel patterns were measured and found to be around 0.6 s for swelling and 2 s for shrinking (for a 60 μm diameter gel) in agreement with the reduced response time expected for scaled down gel patterns. These results suggest the suitability of these gel patterns as valves or actuators in microfluidic devices. Micron-size gel patterns were also incorporated into microfluidic channels thus demonstrating a new approach to create simple, affordable, microfluidic devices, which incorporate “smart” hydrogels as building elements in a simple fashion.     

Velocity-independent microfluidic flow cytometry

Pressure-driven flow in microfluidic channels is characterized by a distribution of velocities. This distribution makes it difficult to implement conventional flow cytometry data analysis. We have demonstrated a method to measure velocity as an independent parameter when performing microfluidic flow cytometry. This method allows velocity-independent analysis of particles such as beads or cells, and allows flow cytometry analysis of extended objects, such as long DNA molecules. It allows accurate flow cytometry in transient and nonuniform flows. This general measurement method could be used in the future to measure the velocity of particles in a variety of existing microfluidic devices without the need for changes in their design.     

Transfer reactions induced by 17, 18O on 12, 13C at c.m. energies between 12.6 TO 14.0 MeV

Angular distributions for one-neutron and many-nucleon transfer reactions were measured in the systems ^{17, 18}O and ^{12, 13}C at c.m. energies between 12.6 to 14.0 MeV. All the cross sections were analyzed in terms of the full-recoil finite-range DWBA model. For multi-nucleon transfer processes an inert cluster transfer was assumed. The sensitivity of DWBA calculations to various parameter sets is discussed, and the effect of the no-recoil approximation on the phase and magnitude of the transfer amplitude is studied. For one-neutron transfer reactions the forward part of the angular distributions was reproduced reasonably well by the DWBA model, yielding satisfactory spectroscopic information. The rise of the cross section at the backward angles was not reproduced by the DWBA model. Possible contributions of cluster-exchange and compound-nucleus reactions are discussed. For multi-nucleon transfer reactions poorer fits were obtained, particularly in the ¹⁸O+¹²C system.     

Determination of the α parameter for the decay Ξ− → Λπ−

The α parameter for Ξ^? → Λπ^? decay has been determined to be α_Ξ^?=?0.462 ± 0.015, from a sample of 1.5 × 10⁵ events recorded in the CERN SPS hyperon beam. This value is 2.5 standard deviations higher than the current world average.     

Formation of inclusion organoactinide complexes with boron-containing macrocycles

Reaction of the organoactinide complexes (C5Me5)2AnMe2 (An = Th, U) with catecholborane yields an inclusion complex where the actinide is encapsulated inside a 15-membered, hexaoxo, trianionic macrocycle built from alternating catechol and catecholborate fragments. In the presence of LiOH, a dimer of two encapsulated actinide macrocycles is formed. The X-ray molecular structure for all the complexes is presented.     

Experimental measurement of the strength of a C alpha-H...O bond in a lipid bilayer

Due to the apolar nature of the lipid bilayer, the weak Calpha-H...O H-bond is thought to contribute significantly toward the stability of transmembrane helical bundles such as glycophorin A (GPA). Here for the first time we measured the strength of such a bond, using vibrational frequency shifts of a dimeric and nondimeric variants of GPA containing a Gly CD2 label. Although the resulting estimated bond strength of 0.88 kcal/mol is relatively weak, several such bonds could contribute significantly toward bundle stabilization.     

Evolution of the mean instantaneous intensity in fibers affected by polarization mode dispersion

It is shown that the mean instantaneous intensity (MII) of a pulse propagating in an optical fiber affected by polarization mode dispersion is related to the frequency autocorrelation of the fiber's Jones matrix through a Fourier transform. A simple derivation of the diffusion equation satisfied by the MII and the autocorrelation function of a Jones matrix is described.     

First observation ofΩ * resonances

In a experiment at the CERN SPS charged hyperon beam using incident Ξ^?, we have obtained evidence for the production of two Ω^* resonances decaying into Ξ^?π⁺ K ^?, with the following parameters:M ₁=2251±12,Γ ₁=48±20 MeV/c², 78 ±23 events, andM ₂=2284±12,Γ ₂=26±23 MeV/c² 45±10 events. The first state is also observed as a 4.2σ effect in a subsample which contains an additionalK ⁺ orK ⁰ in the final state. Production cross sections and branching ratios to Ξ^* K ^? and Ξ^? K ^* are presented.