Construction of a 2,6-difunctionalized 2-methyl-2H-1-benzopyran library by using a solid-phase synthesis protocol

[reaction: see text] A library containing 1200 analogues of 2,6-difunctionalized 2-methyl-2H-1-benzopyran was constructed by using a solid-phase synthesis protocol. Polymer-bound 6-amido-, 6-sulfonamido-, and 6-uredo-functionalized 2-hydroxymethyl-2-methylbenzopyrans 10 were prepared as part of a first-generation diversification step by employing reactions of respective acid halides, sulfonyl chlorides, and isocyanates with the amine precursor 7. Transformations of the resin-bound intermediates 10 by reactions with alkyl and acid halides were then used to produce a diverse series of 2,6-difunctionalized 2-methyl-2H-1-benzopyran analogues 12 and 14.     

Phosphorylation of phospholipase D1 and the modulation of its interaction with RhoA by cAMP-dependent protein kinase

Agents that elevate cellular cAMP are known to inhibit the activation of phospholipase D (PLD). We investigated whether PLD can be phosphorylated by cAMP-dependent protein kinase (PKA) and PKA-mediated phosphorylation affects the interaction between PLD and RhoA, a membrane regulator of PLD. PLD1, but not PLD2 was found to be phosphorylated in vivo by the treatment of dibutyryl cAMP (dbcAMP) and in vitro by PKA. PKA inhibitor (KT5720) abolished the dbcAMP-induced phosphorylation of PLD1, but dibutyryl cGMP (dbcGMP) failed to phosphorylate PLD1. The association between PLD1 and Val14RhoA in an immunoprecipitation assay was abolished by both dbcAMP and dbcGMP. Moreover, RhoA but not PLD1 was dissociated from the membrane to the cytosolic fraction in dbcAMP-treated cells. These results suggest that both PLD1 and RhoA are phosphorylated by PKA and the interaction between PLD1 and RhoA is inhibited by the phosphorylation of RhoA rather than by the phosphorylation of PLD1.     

Synthesis of highly enantioenriched all-carbon quaternary centers: conjugate additions of chiral organolithium nucleophiles to alpha,alpha-dinitrile beta,beta-disubstituted olefins

[reaction: see text]. Highly enantioenriched quaternary centers are obtained by the reaction of chiral lithiated intermediates complexed to (-)-sparteine with tetrasubstituted, alpha,alpha-dinitrile activated olefins. Lithiated N-Boc-N-Aryl benzylamine furnishes products with drs from 78:22 to 95:5, with ers exceeding 94:6. Lithiated N-Boc-N-Aryl allylamine reactants provide enecarbamate products with drs from 55:45 to 99:1, with ers ranging from 87:13 to 97:3.     

Cell lysis on a microfluidic CD (compact disc)

Cell lysis was demonstrated on a microfluidic CD (Compact Disc) platform. In this purely mechanical lysis method, spherical particles (beads) in a lysis chamber microfabricated in a CD, cause disruption of mammalian (CHO-K1), bacterial (Escherichia coli), and yeast (Saccharomyces cerevisiae) cells. Interactions between beads and cells are generated in the rimming flow established inside a partially filled annular chamber in the CD rotating around a horizontal axis. To maximize bead-cell interactions in the lysis chamber, the CD was spun forward and backwards around this axis, using high acceleration for 5 to 7 min. Investigation on inter-particle forces (friction and collision) identified the following parameters; bead density, angular velocity, acceleration rate, and solid volume fraction as having the most significant contribution to cell lysis. Cell disruption efficiency was verified either through direct microscopic viewing or measurement of the DNA concentration after cell lysing. Lysis efficiency relative to a conventional lysis protocol was approximately 65%. In the long term, this work is geared towards CD based sample-to-answer nucleic acid analysis which will include cell lysis, DNA purification, DNA amplification, and DNA hybridization detection.     

Excited-state double proton transfer of 7-azaindole in water nanopools

The excited-state proton-transfer dynamics of 7-azaindole occurring in the water nanopools of reverse micelles has been investigated by measuring time-resolved fluorescence spectra and kinetics, as well as static absorption and emission spectra, with varying water content and isotope. 7-Azaindole molecules are found to exist in the bound-water regions of reverse micelles. The rate constant and the kinetic isotope effect of proton transfer are smaller than those in bulk water although both increase with the size of the water nanopool. The retardation of proton transfer in the bound regions is attributed to the increased free energy of prerequisite solvation to form a cyclically H-bonded 1:1 7-azaindole/water complex.     

Non-newtonian viscosity of polypeptides in the helix–coil transition region

The non-Newtonian intrinsic viscosities [η] of poly(γ-methyl L-glutamate) were measured in the helix–coil transition region under various conditions in this work. The helix content f_H, which represents the degree of conformational transition, was obtained by using a polarimeter. Our experimental results show that the non-Newtonian behavior of the polypeptide is markedly affected by its conformation; i.e., the non-Newtonian effect becomes larger as f_H increases. The effect of external pressure ΔP on [η] was studied carefully; [η] increases with f_H when ΔP < 1.5 psi, but it decreases when ΔP > 1.5 psi and f_H > 0.8. The reason for this result is considered in the text.     

Reversal of facial selectivity in complex Diels-Alder reactions

A complex Diels-Alder reaction between a semi-cyclic diene with allylic silyloxy substituents and a bromo enone presented an unusual diastereoselectivity: attack of the diene occured on its more hindered face, and this reversal of selectivity was shown to be induced by the presence of a bromo substituent in the dienophile.     

Infrared-visible and visible-visible double resonance spectroscopy of 1-hydroxy-9,10-anthraquinone-(H2O)n (n=1,2) complexes

The structures of hydrated 1-hydroxyanthraquinone complexes (1-HAQ), 1-HAQ(H2O)n=1,2, with intramolecular and intermolecular hydrogen bonding interactions were studied using laser spectroscopic methods such as laser induced fluorescence, fluorescence-detected infrared, infrared-visible hole burning, and visible-visible hole burning spectroscopy. In the 1:1 complex 1-HAQ(H2O)1, the water binds to the free carbonyl group of 1-HAQ not associated with intramolecular hydrogen bond. The second water in the 1:2 complex, 1-HAQ(H2O)2, binds to the first water of the 1:1 complex rather than other hydrogen bonding sites of 1-HAQ. A pair of two geometric isomers was produced in a supersonic jet for each of the 1:1 and 1:2 complexes. Both isomers of each complex have the same vibrational spectra in the region of the OH stretching vibration of water, but have different energies for the 0-0 band of vibronic transition due to the asymmetry of the two phenyl rings in 1-HAQ. The 0-0 bands for all four species of 1-HAQ(H2O)n=1,2 were unambiguously assigned by comparing with the results of ab initio calculations, which yielded the structures, vibrational frequencies, and relative energies of the frontier molecular orbitals.     

Electrocapillary at contact: potential-dependent adhesion between a gold electrode and a mica surface

Using the electrochemical surface forces apparatus, we investigated adhesion (from pull-off measurements) between gold and mica as the potential of the gold surface was changed externally. Measurements were performed at different concentrations of KClO(4) in a potential window where the gold electrode is ideally polarizable. At applied potentials where the gold-mica interactions are repulsive, we obtain double layer forces that are predictable by the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory of colloid stability but deviate from the theory at short range. At applied potentials where the gold-mica interactions are attractive, we observed a very strong dependence of adhesion on the applied potential, a result that cannot be directly related to DLVO theory. We show, however, that an approach based on electrocapillary thermodynamics can be employed to model the potential dependence of adhesion seen in our measurements. This electrocapillary approach presents evidence of charging at the gold-mica interface and stresses the relation between the charge within and outside of the contact area.