Insights into 6‐Methylsalicylic Acid Bio‐assembly by Using Chemical Probes

Chemical probes capable of reacting with KS (ketosynthase)‐bound biosynthetic intermediates were utilized for the investigation of the model type I iterative polyketide synthase 6‐methylsalicylic acid synthase (6‐MSAS) in vivo and in vitro. From the fermentation of fungal and bacterial 6‐MSAS hosts in the presence of chain termination probes, a full range of biosynthetic intermediates was isolated and characterized for the first time. Meanwhile, in vitro studies of recombinant 6‐MSA synthases with both nonhydrolyzable and hydrolyzable substrate mimics have provided additional insights into substrate recognition, providing the basis for further exploration of the enzyme catalytic activities.     

Fluorescence properties and photophysics of the sulfoindocyanine Cy3 linked covalently to DNA

The sulfoindocyanine Cy3 is one of the most commonly used fluorescent dyes in the investigation of the structure and dynamics of nucleic acids by means of fluorescence methods. In this work, we report the fluorescence and photophysical properties of Cy3 attached covalently to single-stranded and duplex DNA. Steady-state and time-resolved fluorescence techniques were used to determine fluorescence quantum yields, emission lifetimes, and fluorescence anisotropy decays. The existence of a transient photoisomer was investigated by means of transient absorption techniques. The fluorescence quantum yield of Cy3 is highest when attached to the 5' terminus of single-stranded DNA (Cy3-5' ssDNA), and decreases by a factor of 2.4 when the complementary strand is annealed to form duplex DNA (Cy3-5' dsDNA). Substantial differences were also observed between the 5'-modified strands and strands modified through an internal amino-modified deoxy uridine. The fluorescence decay of Cy3 became multiexponential upon conjugation to DNA. The longest lifetime was observed for Cy3-5' ssDNA, where about 50% of the decay is dominated by a 2.0-ns lifetime. This value is more than 10 times larger than the fluorescence lifetime of the free dye in solution. These observations are interpreted in terms of a model where the molecule undergoes a trans-cis isomerization reaction from the first excited state. We observed that the activation energy for photoisomerization depends strongly on the microenvironment in which the dye is located. The unusually high activation energy measured for Cy3-5' ssDNA is an indication of dye-ssDNA interactions. In fact, the time-resolved fluorescence anisotropy decay of this sample is dominated by a 2.5-ns rotational correlation time, which evidences the lack of rotational freedom of the dye around the linker that separates it from the terminal 5' phosphate. The remarkable variations in the photophysical properties of Cy3-DNA constructs demonstrate that caution should be used when Cy3 is used in studies employing DNA conjugates.     

An end theorem for stratified spaces

We show that a manifold-stratified space X is the interior of a compact manifold-stratified space with boundary if and only if X is tame-ended and a K-theoretic obstruction γ_*(X) vanishes. The obstruction γ_*(X) is a localization of Quinn's mapping cylinder neighborhood obstruction. The main results are Theorem 1.6 and Theorem 1.7 below. In particular, this explains when a G-manifold is the interior of a compact G-manifold with boundary. One of our methods is a new transversality theorem, Theorem 1.16. Oblatum 30-VI-1996 & 21-X-1997 / Published online: 14 January 1999     

Morphology and electrical properties of pulsed-laser deposited Bi2Sr2CaCu2O8+δ films on vicinal substrates

Thin films of Bi₂Sr₂CaCu₂O_8+δ have been grown on vicinal (001) SrTiO₃ substrates by pulsed-laser deposition. Transmission electron microscopy (TEM) and X-ray diffraction reveal well ordered films with the c axis of the film parallel with the c axis of the substrate for miscut angles up to θ_S≈15°. TEM also reveals the step-like film morphology due to step-flow growth. The in-plane and out-of-plane resistivities are independent of film thickness within the range 20–300 nm and agree quite well with Bi₂Sr₂CaCu₂O_8+δ single-crystal data. Received:15 May 2000 / Accepted:17 May 2000 / Published online: 9 August 2000     

Systems-based decomposition schemes for the approximate solution of multi-term fractional differential equations

We give a comparison of the efficiency of three alternative decomposition schemes for the approximate solution of multi-term fractional differential equations using the Caputo form of the fractional derivative. The schemes we compare are based on conversion of the original problem into a system of equations. We review alternative approaches and consider how the most appropriate numerical scheme may be chosen to solve a particular equation.     

Effect of surfactant and solvent properties on the stacking behavior of non-aqueous suspensions of organically modified clays

Montmorillonite clay was treated with quaternary ammonium surfactants with 1-3 long chains of 10-18 carbons to form organoclays which can be suspended in non-aqueous solvents. The effects of surfactant chain length, number of long chains, and the properties of the solvent on the colloidal behavior of the surfactant coated clay plates were studied using small-angle X-ray scattering. The scattering data were modeled using a one-dimensional aggregation theory to describe the stacking of the clay plates. The plates self-organize into stacks with a reproducible basal spacing in the range of 30-50 A, and for each surfactant, the basal spacing falls into one of two preferred distances. We interpret this by considering that the surfactant layer on the clay has two strata, one being the polar near-clay headgroup region and the other the nonpolar alkane chain region. Polar solvents will swell the polar stratum preferentially while nonpolar solvents will swell the nonpolar stratum of the surfactant. As the nonpolar stratum is larger than the polar one, the nonpolar solvents increase the basal spacing between the clay plates more than the polar solvents. The number of long chains on the surfactant does not have an effect on the basal spacing, as the density of surfactant molecules on the surface is low enough to allow the unimpeded swelling of the chains. The one-dimensional aggregation theory can be used to determine the number of plates in a stack, but the effect of changing clay particle size or concentration is not as great as would be expected from this theory. This may be due to the formation of large-scale structures in the suspensions which prevent a true equilibrium stack size being attained.     

Triterpenoids

This review covers the isolation and structure determination of triterpenoids, including squalene derivatives, protostanes, lanostanes, holostanes, cycloartanes, dammaranes, euphanes, tirucallanes, tetranortriterpenoids, quassinoids, lupanes, oleananes, friedelanes, ursanes, hopanes, serratanes and saponins; 305 references are cited.     

The involution on the equivariant Whitehead group

For a finite group G we define an involution on the equivariant Whitehead group given by reversing the direction of an equivariant h-cobordism. It turns out that the involution is not compatible with the splitting of the equivariant Whitehead group into a direct sum of algebraic Whitehead groups, certain correction terms involving the transfer maps of the normal sphere bundles of the various fixed point sets come in. However, if the group has odd order, these transfer maps all vanish. We prove a duality formula for a G-homotopy equivalence (f f): (M; M) (N, N) relating the equivariant Whitehead torsion of f and (f,f).     

On-line solution stability determination of pharmaceuticals by capillary electrophoresis

Summary Good agreement between the impurity levels in a batch of a related impurity of ranitidine were obtained by CE and HPLC. A solution of the impurity was positioned on the CE autosampler and analysed sequentially. The extent of degradation was monitored by loss of main peak and the formation of two principal degradation products. It was found that after 9.25 hours only 2% area/area of the original impurity remained. Buffering of the sample solution to pH 7 was shown to minimise this degradation.Unattended in-situ stability testing of an solution of the impurity in water was performed by CE.