Incorporation of a DNAzyme into Au-coated nanocapillary array membranes with an internal standard for Pb(ii) sensing

A Pb(ii)-specific DNAzyme has been successfully incorporated into Au-coated polycarbonate track-etched (PCTE) nanocapillary array membranes (NCAMs) by thiol-gold immobilization. Incorporation of the DNAzyme into the membrane provides a substrate-bound sensor using a novel internal control methodology for fluorescence-based detection of Pb(ii). A non-cleavable substrate strand, identical to the cleavable DNAzyme substrate strand except the RNA-base is replaced by the corresponding DNA-base, is used for ratiometric comparison of intensities. The cleavable substrate strand is labeled with fluorescein, and the non-cleavable strand is labeled with a red fluorophore (Cy5 or Alexa 546) for detection after release from the membrane surface. This internal standard based ratiometric method allows for real-time monitoring of Pb(ii)-induced cleavage, as well as standardizing variations in substrate size, solution detection volume, and monolayer density. The result is a Pb(ii)-sensing structure that can be stored in a prepared state for 30 days, regenerated after reaction, and detect Pb(ii) concentrations as low as 17 nM (3.5 ppb).     

Stacked conjugated oligomers as molecular models to examine interchain interactions in conjugated materials

Previous studies of the redox states of linear conjugated oligomers as models for polarons and bipolarons in conjugated polymers do not fully address the influence of intermolecular interactions on the electronic structure of conjugated systems in the solid state. Fusion of oligothiophenes onto a bicyclo[4.4.1]undecane core holds the conjugated oligomers in a permanent cofacial stack. One- and two-electron oxidation of the stacked oligomers affords mono(radical cation)s and dications that serve as models for polarons and bipolarons in p-doped conjugated polymers and demonstrates the effect of pi-stacking on the electronic structure of these species.     

Microfluidic technologies as platforms for performing quantitative cellular analyses in an in vitro environment

Quite often, important cellular events occur in environments that are either not amenable to implanted sensors or other types of molecular probes. In such cases, a viable alternative to taking the sensor or probe to the biological sample of interest is to bring the sample of interest out of its natural environment to one that is more conducive to the measurement scheme. The disadvantage of the latter approach is that the sample may not behave in the same manner in vitro as it does in vivo, or that the agonists and other stimuli to which the sample is subjected to in vivo are no longer present. In this Tutorial Review, the authors attempt to provide some guidance, based on their own experiences and those of other scientists, to performing cellular measurements in a quantitative manner under in vitro conditions. Due to the expansive literature on analyses involving cells, the authors have limited this Tutorial Review to those methods involving microfluidic technologies, both in microbore tubing and in microfabricated channels. Initial reports of analyses involving cells in microbore tubing were first reported nearly two decades ago, while those in microfabricated fluidic devices appeared over a decade ago. However, more recently, the complexity of cell analyses using fabricated microfluidic devices (as opposed to microbore tubing) has increased due in part to the improvements in fabrication technologies, fluid handling and delivery capabilities, advances in coatings of the channels within the microfluidic device, and integrated detection schemes. Examples of cellular analyses in microbore tubing and in fabricated microfluidic devices will be given, as well as associated advantages and challenges. Finally, the authors' thoughts on cellular analyses are presented here using the classical steps in an analysis as a guide.     

Synthesis of organically templated nanoporous Tin(II/IV) phosphate for radionuclide and metal sequestration

Nanoporous tin(II/IV) phosphate materials, with spherical morphology, have been synthesized using cetyltrimethylammonium chloride [CH3(CH2)15N(CH3)3Cl] as the surfactant. The structure of the material is stable at 500 degrees C; however, partial oxidation of the material occurs with redox conversion of Sn2+ to Sn4+, resulting in a mixed Sn(II)/Sn(IV) material. Preliminary batch contact studies were conducted to assess the effectiveness of nanoporous tin phosphate, NP-SnPO, in sequestering redox-sensitive metals and radionuclides, technetium(VII), neptunium(V), thorium(IV), and a toxic metal, chromium(VI), from aqueous matrixes. Results indicate that tin(II) phosphate removed >95% of all contaminants investigated from solution.     

Clay-fulleropyrrolidine nanocomposites

In this work, we describe the insertion of a water-soluble bisadduct fulleropyrrolidine derivative into the interlayer space of three layered smectite clays. The composites were characterized by a combination of powder X-ray diffraction, transmission electron microscopy, X-ray photoemission and FTIR spectroscopies, and laser flash photolysis measurements. The experiments, complemented by computer simulations, give insight into the formation process, structural details, and properties of the fullerene/clay nanocomposites. The reported composite materials constitute a new hybrid system, where C(60) differs from its crystals or its solutions, and open new perspectives for the design and construction of novel C(60)-based organic/clay hybrid materials.     

Asymmetric Michael addition of nitroalkanes to nitroalkenes catalyzed by C2-symmetric tridentate bis(oxazoline) and bis(thiazoline) zinc complexes

The first asymmetric synthesis of 1,3-dinitro compounds through Michael addition of nitroalkanes to nitroalkenes catalyzed by C2-symmetric chiral tridentate bis(oxazoline) and bis(thiazoline) zinc complexes was achieved with high enantioselectivities (up to 95% ee).     

CB11Me11 boronium ylides: carba-closo-dodecaboranes with a naked boron vertex

In pentane solution, 2 equiv of the icosahedral CB(11)Me(12)(*) radical cleaves the Si-Si bond of hexaalkyldisilanes by boron to silicon methyl transfer with formation of 2 equiv of methyltrialkylsilanes. The loss of a methyl radical converts the CB(11)Me(12)(*) radical into an internally charge-compensated "boronium ylide" CB(11)Me(11) with a naked vertex, which can be formally viewed as a deprotonated hypercloso carborane. It has been isolated as an air-sensitive solid, stable only below approximately -60 degrees C. The naked vertex appears to be in position 12 since the material reacts instantaneously with alcohols and ethers to form the 12-alkoxy anions 12-CB(11)Me(11)OR. It reacts with many other nucleophiles to yield more complex mixtures containing similar products. DFT calculations for the four CB(11)Me(11) isomers give closed-shell ground-state electronic structures. For the isomer with naked vertex 12, a DFT computational search failed to reveal any skeletal dimers, apparently due to excessive methyl-methyl repulsions, and only a cyclic dimer bound through weak interactions of one of the 7-methyl hydrogen atoms on each cage with the empty exocyclic orbitals on B12 of the other cage was found. Natural hybrid orbital populations suggest that the three possible isomers of monomeric boronium ylides are close to true singlet ylides, with triplet states approximately 50 kcal/mol higher. The calculated electronic structure of the carbonium ylide is close to a singlet carbene, with a triplet state approximately 16 kcal/mol higher. An attempted preparation of Me(3)C(+)CB(11)Me(12)(-) yielded neopentane and products consistent with a sequential loss of methyl groups from the carborane cage with a transient formation of similar boronium ylides. Probable mechanisms of these methyl transfer reactions are considered, and the possibly quite general role of "ylide" structures in Lewis acid induced substitution reactions on the boron vertices of carboranes and boranes is noted.     

Size and charge effects on the binding of CO to late transition metal clusters

We report on the size and charge dependence of the C-O stretching frequency, nu(CO), in complexes of CO with gas phase anionic, neutral, and cationic cobalt clusters (Co(n)CO(-0+)), anionic, neutral, and cationic rhodium clusters (Rh(n)CO(-0+)), and cationic nickel clusters (Ni(n)CO(+)) for n up to 37. We develop models, based on the established vibrational spectroscopy of organometallic carbonyl compounds, to understand how cluster size and charge relate to nu(CO) in these complexes. The dominating factor is the available electron density for backdonation from the metal to the CO pi* orbital. Electrostatic effects play a significant but minor role. For the charged clusters, the size trends are related to the dilution of the charge density at the binding site on the cluster as n increases. At large n, nu(CO) approaches asymptotes that are not the same as found for nu(CO) on the single crystal metal surfaces, reflecting differences between binding sites on medium sized clusters and the more highly coordinated metal surface sites.     

Photoelectron imaging of I2- at 5.826 eV

We report the anion photoelectron spectrum of I2- taken at 5.826 eV detachment energy using velocity mapped imaging. The photoelectron spectrum exhibits bands resulting from transitions to the bound regions of the X 1Sigmag+(0g+), A' 3Piu(2u), A 3Piu(1u), and B 3Piu(0u+) electronic states as well as bands resulting from transitions to the repulsive regions of several I2 electronic states: the B' 3Piu(0u-), B" 1Piu(1u), 3Pig(2g), a 3Pig(1g), 3Pig(0g-), and C 3Sigmau+(1u) states. We simulate the photoelectron spectrum using literature parameters for the I2- and I2 ground and excited states. The photoelectron spectrum includes bands resulting from transitions to several high-lying excited states of I2 that have not been seen experimentally: 3Pig(0g-), 1Pig3(1g), 1 3Sigmag-3(0g+), and the 1Sigmag-3(0u-) states of I2. Finally, the photoelectron spectrum at 5.826 eV allows for the correction of a previous misassignment for the vertical detachment energy of the I2 B 3Piu(0u+) state.     

Simple estimation of absolute free energies for biomolecules

One reason that free energy difference calculations are notoriously difficult in molecular systems is due to insufficient conformational overlap, or similarity, between the two states or systems of interest. The degree of overlap is irrelevant, however, if the absolute free energy of each state can be computed. We present a method for calculating the absolute free energy that employs a simple construction of an exactly computable reference system which possesses high overlap with the state of interest. The approach requires only a physical ensemble of conformations generated via simulation and an auxiliary calculation of approximately equal central-processing-unit cost. Moreover, the calculations can converge to the correct free energy value even when the physical ensemble is incomplete or improperly distributed. As a "proof of principle," we use the approach to correctly predict free energies for test systems where the absolute values can be calculated exactly and also to predict the conformational equilibrium for leucine dipeptide in implicit solvent.