Use of an automated image processing program to quantify recombinant adenovirus particles.

Electron microscopy has a pivotal role as an analytical tool in pharmaceutical research. However, digital image data have proven to be too large for efficient quantitative analysis. We describe here the development and application of an automated image processing (AIP) program that rapidly quantifies shape measurements of recombinant adenovirus (rAd) obtained from digitized field emission scanning electron microscope (FESEM) images. The program was written using the macro-recording features within Image-Pro Plus software. The macro program, which is linked to a Microsoft Excel spreadsheet, consists of a series of subroutines designed to automatically measure rAd vector objects from the FESEM images. The application and utility of this macro program has enabled us to rapidly and efficiently analyze very large data sets of rAd samples while minimizing operator time.     

Cyclic Voltammetry in a Perfluorocarbon Emulsion Blood Substitute

Cyclic voltammetry in a perfluorocarbon emulsion based blood substitute (PEBS) was evaluated. The intent was to determine how PEBS affects the voltammetry of four representative electroactive compounds: potassium ferricyanide, ruthenium(II) trisbipyridine (rutris), hydroquinone, and 2,6‐dichloroindophenol (DCIP). Voltammograms in 0–20% PEBS‐Tris buffer mixtures are affected by PEBS, but reasonably well‐defined voltammograms are obtained in as much as 20% v/v. This report also shows that PEBS has only a small effect on amperometric detection of the reaction between DCIP and nicotinamide adenine dinucleotide (NADH), which has importance in clinical homogeneous immunoassays. Results support continued exploration of voltammetry/amperometry for quantitative analysis in this medium.     

General deoxyribozyme-catalyzed synthesis of native 3'-5' RNA linkages

An elusive goal for nucleic acid enzymology has been deoxyribozymes that ligate RNA rapidly, sequence-generally, with formation of native 3'-5' linkages, and in preparatively useful yield. Using in vitro selection, we have identified Mg2+- and Zn2+-dependent deoxyribozymes that simultaneously fulfill all four of these criteria. The new deoxyribozymes operate under practical incubation conditions and have modest RNA substrate sequence requirements, specifically D downward arrowRA for 9DB1 and A downward arrowR for 7DE5 (D = A, G, or U; R = A or G). These requirements are comparable to those of deoxyribozymes such as 10-23 and 8-17, which are already widely used as biochemical tools for RNA cleavage. We anticipate that the 9DB1 and 7DE5 deoxyribozymes will find immediate practical application for RNA ligation.     

The Bohlmann-Rahtz route to functionalised pyridine scaffolds and their use in library synthesis

The Bohlmann-Rahtz reaction has been used to prepare 2,3,6-trisubstituted pyridines suitable for use in library synthesis. The synthesis of piperidine substituted nicotinic acid derivative 9 has been optimised and carried out on a large scale to give ca. 500 g of scaffold which was used in the generation of the pyridine library 11.     

Structural properties of ultra-small thorium and uranium dioxide nanoparticles embedded in a covalent organic framework

We report the structural properties of ultra-small ThO₂ and UO₂ nanoparticles (NPs), which were synthesized without strong binding surface ligands by employing a covalent organic framework (COF-5) as an inert template. The resultant NPs were used to observe how structural properties are affected by decreasing grain size within bulk actinide oxides, which has implications for understanding the behavior of nuclear fuel materials. Through a comprehensive characterization strategy, we gain insight regarding how structure at the NP surface differs from the interior. Characterization using electron microscopy and small-angle X-ray scattering indicates that growth of the ThO₂ and UO₂ NPs was confined by the pores of the COF template, resulting in sub-3 nm particles. X-ray absorption fine structure spectroscopy results indicate that the NPs are best described as ThO₂ and UO₂ materials with unpassivated surfaces. The surface layers of these particles compensate for high surface energy by exhibiting a broader distribution of Th–O and U–O bond distances despite retaining average bond lengths that are characteristic of bulk ThO₂ and UO₂. The combined synthesis and physical characterization efforts provide a detailed picture of actinide oxide structure at the nanoscale, which remains highly underexplored compared to transition metal counterparts.

ThO₂ and UO₂ nanoparticles synthesized using a COF-5 template exhibit unpassivated surfaces and provide insight into nanoscale properties of actinides.     

Synthesis of the B-ring of FR182877. Investigation of the reactions of 6-fumaryl 1,3,8-nonatrienes

Investigation of the intramolecular Diels-Alder reactions of 6-fumaryl 1,3,8-nonatrienes, substituted at the C5 by a vinyl group, to afford the B-ring of FR182877 are reported. The synthesis of the required 1,3,8-nonatriene was achieved quickly and in high yield. 6-Fumaryl 1,3,8-nonatrienes substituted at the C5 by a vinyl group were found to undergo competing tandem sigmatropic rearrangement/Diels-Alder cyclisation when heated under standard Diels-Alder cyclisation conditions. This rearrangement became the exclusive pathway when the reaction was performed in the presence of a Lewis acid. As expected from modeling studies, the major intramolecular Diels-Alder cyclisation product was the desired exo-trans adduct, which was required for the synthesis of FR182877. Intrigued by the rearrangements, a number of alterations were made to the 1,3,8-nonatriene. Replacement of the fumaryl group by an acetyl group resulted in the diminished reactivity of the 1,3,8-nonatriene with neither rearrangements nor cycloadditions observed. Variation of the C5 substituent was found to be very important in determining the π-diastereoselectivity of the Diels-Alder cyclisation.     

Ultraviolet photochemistry of ethane: implications for the atmospheric chemistry of the gas giants

Chemical processing in the stratospheres of the gas giants is driven by incident vacuum ultraviolet (VUV) light. Ethane is an important constituent in the atmospheres of the gas giants in our solar system. The present work describes translational spectroscopy studies of the VUV photochemistry of ethane using tuneable radiation in the wavelength range 112 ≤ λ ≤ 126 nm from a free electron laser and event-triggered, fast-framing, multi-mass imaging detection methods. Contributions from at least five primary photofragmentation pathways yielding CH₂, CH₃ and/or H atom products are demonstrated and interpreted in terms of unimolecular decay following rapid non-adiabatic coupling to the ground state potential energy surface. These data serve to highlight parallels with methane photochemistry and limitations in contemporary models of the photoinduced stratospheric chemistry of the gas giants. The work identifies additional photochemical reactions that require incorporation into next generation extraterrestrial atmospheric chemistry models which should help rationalise hitherto unexplained aspects of the atmospheric ethane/acetylene ratios revealed by the Cassini–Huygens fly-by of Jupiter.

The vacuum ultraviolet photodissociation dynamics of ethane provide clues for modelling the atmospheric chemistry of the gas giants.     

INDUCTION OF CHROMOSOME ABERRATIONS IN ICR 2A FROG CELLS EXPOSED TO265–313 nm MONOCHROMATIC ULTRAVIOLET WAVELENGTHS and PHOTOREACTIVATING LIGHT

Abstract— Exposure of ICR 2A cells to either 265, 289, 302 or 313 nm monochromatic UV wavelengths caused the induction of chromosome aberrations with chromatid gaps and breaks being the most common type of aberration detected. Treatment of U V-irradiated cells with photoreactivating light (PRL) resulted in a lower yield of aberrations demonstrating that pyrimidine dimers are involved in the formation of chromosome aberrations induced by the UV wavelengths tested. However, the decrease in the level of aberrations resulting from PRL treatment of 313 nm-irradiated cells was significantly less than for the other wavelengths indicating that non-dimer photoproducts may have played an important additional role in the induction of chromosome aberrations by this UV wavelength.     

Determination of the size and composition of multicomponent ethanol/water droplets by cavity-enhanced Raman scattering

Cavity-enhanced Raman scattering is used to determine the size and composition of multicomponent ethanol/water droplets in the concentration range 7.5–19% ethanol by volume. Under the experimental conditions presented here, the integrated CERS signal from ethanol shows an exponential increase with increase in ethanol concentration when compared with the integrated intensity of the water band. The calibration is shown to be invariant with particle size over the droplet radius range 20–35 μm. In addition to providing a method for determining particle size and composition, initial studies show that the evaporation dynamics of these multicomponent droplets can be probed by CERS.