Phase behavior and 3D structure of strongly attractive microsphere-nanoparticle mixtures

We investigate the phase behavior and 3D structure of strongly attractive mixtures of silica microspheres and polystyrene nanoparticles. These binary mixtures are electrostatically tuned to promote a repulsion between like-charged (microsphere-microsphere and nanoparticle-nanoparticle) species and a strong attraction between oppositely charged (microsphere-nanoparticle) species. Using confocal fluorescence scanning microscopy, we directly observe the 3D structure of colloidal phases assembled from these mixtures as a function of varying composition. In the absence of nanoparticle additions, the charged-stabilized microspheres assemble into a polycrystalline array upon sedimentation. With increasing nanoparticle volume fraction, nanoparticle bridges form between microspheres, inducing their flocculation. At even higher nanoparticle volume fractions, the microspheres become well coated with nanoparticles, leading to their charge reversal and subsequent restabilization. We demonstrate how this fluid-gel-fluid transition can be utilized to control the morphology of the colloidal phases formed under gravity-driven sedimentation.     

Behavior of M[Al2Me6N3] (M=K,Rb, Cs) with aromatic solvents and the crystal structures of Cs[Al2Me6N3]·2p-xylene and [K·dibenzo-18-crown-6] [Al2Me6N3]· 1.5(1-methylnaphthalene)

The title compounds were synthesized by the addition of AlMe₃ to the corresponding azide suspended in an aromatic solvent. Both products were obtained as air-sensitive colorless crystals. Cs[Al₂Me₆N₃]·2p-xylene crystallizes in the monoclinic space groupC2/m witha=19.143(6),b=16.227(6),c=10.392(5) Å, =114.06(2)^o, and _calc = 1.20 g cm^–3 forZ=4. Refinement led to a conventionalR value of 0.037 for 2179 observed reflections. The cesium atom resides on a mirror plane, and the anion is disordered about a twofold axis. Thep-xylene molecules sandwich the cesium ion.[K·dibenzo-18-crown-6] [Al_Me6N₃]·1.5(1-methylnaphthalene) crystallizes in the monoclinic space groupP2₁/c witha=14.176(5),b=13.021(5),c=25.324(8) Å, =98.23(4)⁰, and _calc = 1.08 g cm^–3 forZ=4. The finalR value was 0.132 for 1402 observed reflections. One of the 1-methylnaphthalene molecules is disordered about a center of inversion and interacts with the potassium ion. The other solvent molecule is found roughly in layers in the lattice and also exhibits disorder of the methyl substituent. For both title compounds the AlMe₃ groups of the anion exhibit a staggered (C _s) conformation. Supplementary Data relating to this article are deposited with the British Library as Supplementary Publication No. SUP 82015 (32 pages).     

Transient absorption studies and numerical modeling of iodine photoreduction by nanocrystalline TiO2 films

We have used transient absorption spectroscopy to study the reaction between photogenerated electrons in a dye-free nanocrystalline titanium dioxide film and an iodine/iodide redox couple. Recombination kinetics was measured by recording the transient optical signal following band gap excitation by a UV laser pulse. In the presence of a methanol hole scavenger in the electrolyte, a long-lived (0.1-1 s) red/infrared absorbance is observed and assigned to photogenerated electrons forming Ti(3+) species. In the presence of iodine and excess iodide in the electrolyte, the signal decays on a millisecond-microsecond time scale, assigned to reduction of the redox couple by photogenerated electrons in the TiO(2). The electron lifetime decreases inversely with increasing iodine concentration, indicating that the back reaction is first order in [I(2)]. No evidence for I(2)(-) is observed, indicating that the reaction mechanism does not involve the formation of I(2)(-) as an intermediate. The shape of the kinetics evolves from monoexponential at low [I(2)] to stretched-exponential as [I(2)] increases. A Monte Carlo continuous-time random walk model is implemented to simulate the kinetics and its [I(2)] dependence and used to address the order of the recombination reaction with respect to electron density, n. The model incorporates the diffusion of oxidized species from the electrolyte toward the TiO(2) surface as well as electron trapping and transport in the TiO(2). In the limit of low [I(2)], the monoexponential kinetics is explained by the recombination reaction being rate limited by the diffusion of the oxidized species in the electrolyte. The stretched-exponential behavior at high [I(2)] can be explained by the reaction being rate limited by the transport of electrons through a distribution of trap states toward reactive sites at the TiO(2)-electrolyte interface, similar to the mechanism proposed previously for the kinetics of electron-dye cation recombination. Such trap-limited recombination can also explain the superlinear dependence of electron recombination rate on electron density, which has been reported elsewhere, without the need for a reaction mechanism that is second order in n. In contrast, a second-order reaction mechanism in a trap-free medium cannot explain the observed kinetics, although a second-order mechanism incorporating electron trapping cannot be conclusively ruled out by the data. We propose that the most likely reaction scheme, that is first order in both [I(2)] and n, is the dissociative reduction of I(2) onto the metal oxide surface, followed by a second electron reduction of the resulting adsorbed iodine radical, and that empirical second-order behavior of the electron lifetime is most likely explained by electron trapping rather than by a second-order recombination mechanism.     

Crystal structures, reactivity and inferred acylation transition states for 2'-amine substituted RNA

Ribose 2'-amine substitutions are broadly useful as structural probes in nucleic acids. In addition, structure-selective chemical reaction at 2'-amine groups is a robust technology for interrogating local nucleotide flexibility and conformational changes in RNA and DNA. We analyzed crystal structures for several RNA duplexes containing 2'-amino cytidine (C(N)) residues that form either C(N)-G base pairs or C(N)-A mismatches. The 2'-amine substitution is readily accommodated in an A-form RNA helix and thus differs from the C2'-endo conformation observed for free nucleosides. The 2'-amide product structure was visualized directly by acylating a C(N)-A mismatch in intact crystals and is also compatible with A-form geometry. To visualize conformations able to facilitate formation of the amide-forming transition state, in which the amine nucleophile carries a positive partial charge, we analyzed crystals of the C(N)-A duplex at pH 5, where the 2'-amine is protonated. The protonated amine moves to form a strong electrostatic interaction with the 3'-phosphodiester. Taken together with solution-phase experiments, 2'-amine acylation is likely facilitated by either of two transition states, both involving precise positioning of the adjacent 3'-phosphodiester group.     

Detection of erythrocyte membrane structural abnormalities in lecithin: cholesterol acyltransferase deficiency using a spin label approach.

The membrane fluidity of erythrocytes from patients with Lecithin: cholesterol acyltransferase (LCAT) deficiency was studied by means of electron spin resonance. The temperature dependence of the separation of the outer extrema of the spectra of 2-(3-carboxy-propyl)-4,4-dimethyl, 2-tridecyl-3-oxazolidinyloxyl spin probe was monitored for normal, presumed carrier and clinically affected subjects. The temperature profile of controls was significantly different from that of the presumed carriers and the clinically affected individuals. The results show that the compositional abnormalities previously noted in erythrocyte membranes from patients with LCAT deficiency are associated with alterations in the physiocochemical state of the membrane. An investigation of the spectral lineshapes below 10 degrees C allowed a distinction to be made at the membrane level between clinically affected subjects and clinically normal heterozygous carriers. Alterations in the temperature dependence of elec-ron spin resonance parameters may provide a sensitive index of red cell membrane alterations in pathological states of generalized membrane involvement.     

Sequence‐Defined Oligomers from Hydroxyproline Building Blocks for Parallel Synthesis Applications

The functionality of natural biopolymers has inspired significant effort to develop sequence‐defined synthetic polymers for applications including molecular recognition, self‐assembly, and catalysis. Conjugation of synthetic materials to biomacromolecules has played an increasingly important role in drug delivery and biomaterials. We developed a controlled synthesis of novel oligomers from hydroxyproline‐based building blocks and conjugated these materials to siRNA. Hydroxyproline‐based monomers enable the incorporation of broad structural diversity into defined polymer chains. Using a perfluorocarbon purification handle, we were able to purify diverse oligomers through a single solid‐phase extraction method. The efficiency of synthesis was demonstrated by building 14 unique trimers and 4 hexamers from 6 diverse building blocks. We then adapted this method to the parallel synthesis of hundreds of materials in 96‐well plates. This strategy provides a platform for the screening of libraries of modified biomolecules.     

Slow conformational dynamics at C2'-endo nucleotides in RNA

RNA molecules undergo local conformational dynamics on timescales spanning picoseconds to minutes. Slower local motions have the greater potential to govern RNA folding, ligand recognition, and ribonucleoprotein assembly reactions but are difficult to detect in large RNAs with complex structures. RNA SHAPE chemistry employs acylation of the ribose 2'-hydroxyl position to measure local nucleotide flexibility in RNA and is well-characterized by a mechanism in which each nucleotide samples unreactive (closed) and reactive (open) states. We monitor RNA conformational dynamics over distinct time domains by varying the electrophilicity of the acylating reagent. Select C2'-endo nucleotides are nonreactive toward fast reagents but reactive toward slower SHAPE reagents in both model RNAs and in a large RNA with a tertiary fold. We conclude, first, that the C2'-endo conformation by itself does not govern SHAPE reactivity. However, some C2'-endo nucleotides undergo extraordinarily slow conformational changes, on the order of 10(-4) s(-1). Due to their distinctive local dynamics, C2'-endo nucleotides have the potential to function as rate-determining molecular switches and are likely to play central, currently unexplored, roles in RNA folding and function.     

Stereoselective synthesis of (2S,3R)- and (2R,3S)-iodoreboxetine; potential SPECT imaging agents for the noradrenaline transporter

With the aim of developing a new SPECT imaging agent for the noradrenaline transporter, a twelve-step stereoselective synthesis of iodinated analogues of (2S,3R)- and (2R,3S)-reboxetine has been achieved from 4-bromobenzaldehyde. The key steps involve a Sharpless asymmetric epoxidation to establish the stereogenic centres and a copper catalysed aromatic halogen exchange reaction to introduce the key iodine atom. In vitro testing of these compounds using a [(3)H]nisoxetine displacement assay with homogenised rat brain shows both compounds to have significant affinity, with K(i) values of 320.8 nM and 58.2 nM for (2S,3R)- and (2R,3S)-iodoreboxetine respectively.     

Suzuki-Miyaura approach to JNJ-26076713, an orally active tetrahydroquinoline-containing alphaVbeta3/alphaVbeta5 integrin antagonist. enantioselective synthesis and stereochemical studies

An improved scale-up synthesis was required for the alpha(V)beta(3)/alpha(V)beta(5) integrin antagonist 1, which had demonstrated oral efficacy in eye disease models of angiogenesis and vascular permeability. A stereodefined, quinoline-substituted, unsaturated ester was conveniently prepared by a Suzuki-Miyaura coupling to facilitate exploration of multiple methods of asymmetric reduction. The catalytic chiral hydrogenation of the corresponding unsaturated acid (Z-5b) with a ruthenium-based metal precursor and the (R)-XylPhanePhos ligand proved particularly efficient and economical. The resulting (3S)-quinoline-containing intermediate was reduced to an equal mixture of tetrahydroquinoline diastereomers. The undesired diastereomer could be recycled to the desired one by an oxidation/reduction protocol. The absolute stereochemistry of 1 was established as 3S,3'S by a combination of X-ray diffraction and chemical means.