Extending the biocatalytic scope of regiocomplementary flavin-dependent halogenase enzymes

Flavin-dependent halogenases are potentially valuable biocatalysts for the regioselective halogenation of aromatic compounds. These enzymes, utilising benign inorganic halides, offer potential advantages over traditional non-enzymatic halogenation chemistry that often lacks regiocontrol and requires deleterious reagents. Here we extend the biocatalytic repertoire of the tryptophan halogenases, demonstrating how these enzymes can halogenate a range of alternative aryl substrates. Using structure guided mutagenesis we also show that it is possible to alter the regioselectivity as well as increase the activity of the halogenases with non-native substrates including anthranilic acid; an important intermediate in the synthesis and biosynthesis of pharmaceuticals and other valuable products.     

The structure and dynamics of reacting plane mixing layers

The reacting two-dimensional plane mixing layer has been studied in two configurations: a rearward facing step and a two-stream mixing layer. Observations have been made of the steady state behavior, and the unsteady behavior when the flow was forced by a specific acoustic frequency. The steady behavior of the mean properties of the reacting flows is similar to that of non-reacting free shear flows except for the global effects of thermodynamic property changes. The structure of these flows is qualitatively similar to that of non-reacting flows. Vortices form by the two-dimensional Kelvin-Helmholtz instability and grow by subharmonic combination until the mixing layer interacts with the walls. Entrainment is dominated by the two-dimensional vortex motion. Three-dimensional instabilities give rise to secondary vortices which are coherent over several Kelvin-Helmholtz structures and dominate the fine scale mixing process. The mixing transition corresponds to a loss of coherence in the layer. Unsteady behavior occurs when there are resonant interactions with the Kelvin-Helmholtz instability or the instability associated with the recirculation vortex in the rearward facing step flow. Modeling efforts are reported which show promise of simulating the essential features of plane mixing layers.A version of this paper was presented at the ASME Winter Annual Meeting of 1984 and printed in AMD-Vol. 66     

Experimental criteria for the determination of fractal parameters of premixed turbulent flames

The influence of spatial resolution, digitization noise, the number of records used for averaging, and the method of analysis on the determination of the fractal parameters of a high Damköhler number, methane/air, premixed, turbulent stagnation-point flame are investigated in this paper. The flow exit velocity was 5 m/s and the turbulent Reynolds number was 70 based on a integral scale of 3 mm and a turbulent intensity of 7%. The light source was a copper vapor laser which delivered 20 nsecs, 5 mJ pulses at 4 kHz and the tomographic cross-sections of the flame were recorded by a high speed movie camera. The spatial resolution of the images is 155 × 121 m/pixel with a field of view of 50 × 65 mm. The stepping caliper technique for obtaining the fractal parameters is found to give the clearest indication of the cutoffs and the effects of noise. It is necessary to ensemble average the results from more than 25 statistically independent images to reduce sufficiently the scatter in the fractal parameters. The effects of reduced spatial resolution on fractal plots are estimated by artificial degradation of the resolution of the digitized flame boundaries. The effect of pixel resolution, an apparent increase in flame length below the inner scale rolloff, appears in the fractal plots when the measurent scale is less than approximately twice the pixel resolution. Although a clearer determination of fractal parameters is obtained by local averaging of the flame boundaries which removes digitization noise, at low spatial resolution this technique can reduce the fractal dimension. The degree of fractal isotropy of the flame surface can have a significant effect on the estimation of the flame surface area and hence burning rate from two-dimensional images. To estimate this isotropy a determination of the outer cutoff is required and three-dimensional measurements are probably also necessary.     

Velocity statistics along the stagnation line of an axi- symmetric stagnating turbulent flow

 Velocity statistics along the stagnation line of an axi-symmetric wall stagnating turbulent flow are studied experimentally. A low turbulence, uniform air flow from a nozzle type air supply with an exit diameter of 50 mm stagnates at a wall located 50 mm downstream. A flow velocity is set to 3 m/s, 10 mm downstream from the exit of the air supply. Instantaneous values of streamwise and radial velocities are measured by laser-Doppler velocimetry. The turbulence level in the air flow is changed by use of turbulence generator. When the turbulence generator is not installed in the air supply, the mean velocity profile in the streamwise direction fits well with that of a laminar viscous flow with the rms value of velocity fluctuations low near the wall. With the turbulence generator installed, a significant turbulence structure appears near the wall. When the wall is approached, the rms value of velocity fluctuations in the streamwise direction decreases monotonically while the profile of the rms value in the radial direction reaches a maximum near the wall. The increase in the rms value of velocity fluctuations in the radial direction near the wall is attributed to the bi-modal histogram of the fluctuating velocity in the radial direction. Near the wall, the instantaneous stagnation streamline fluctuates and the probability of the mean location of the stagnation point reaches a maximum not at the stagnation line but on a circle around the stagnation line, resulting in the bi-modal histogram. Turbulence statistics, the rms value of velocity fluctuation and the turbulent kinetic energy, can be normalized successfully by similarity parameters based on the strain rate and the reference turbulent kinetic energy introduced by Champion and Libby. Received: 7 April 1995/Accepted: 27 September 1996     

β-Ketoiminato Iridium(III) Organometallic Complexes: Selective Cytotoxicity towards Colorectal Cancer Cells HCT116 p53-/-

This report presents a new library of organometallic iridium(III) compounds of the type [Cp*IrCl(L)] (Cp*=pentamethylcyclopentadienyl and L=a functionalized β-ketoiminato ligand) showing moderate to high cytotoxicity against a range of cancer cell lines. All compounds show increased activity towards colorectal cancer, with preferential activity observed against the immortalized p53-null colorectal cell line, HCT116 p53-/-, with sensitivity factors (SF) up to 26.7. Additionally, the compounds have excellent selectivity for cancerous cells when tested against normal cell types, with selectivity ratios (SR) up to 35.6, contrary to that of cisplatin, which is neither selective nor specific for cancerous cells (SF=0.43 and SR=0.7–2.3). This work provides a preliminary understanding of the cytotoxicity of iridium compounds in the absence of p53 and has potential applications in treatment of cancers for which the p53 gene is absent or mutant.     

Iron-Catalyzed Halogen Exchange of Trifluoromethyl Arenes**

The facile production of ArCF₂X and ArCX₃ from ArCF₃ using catalytic iron(III)halides is reported, which constitutes the first iron-catalyzed halogen exchange for non-aromatic C−F bonds. Theoretical calculations suggest direct activation of C−F bonds by iron coordination. ArCX₃ and ArCF₂X products of the reaction are synthetically valuable due to their diversification potential. In particular, chloro- and bromodifluoromethyl arenes (ArCF₂Cl, ArCF₂Br respectively) provide access to a myriad of difluoromethyl arene derivatives (ArCF₂R). To optimize for mono-halogen exchange, a statistical method called Design of Experiments was used. Optimized parameters were successfully applied to electron rich and electron deficient aromatic substrates, and to the late stage diversification of flufenoxuron, a commercial insecticide. These methods are highly practical, being run at convenient temperatures and using inexpensive common reagents.     

Reduced field-of-view diffusion-weighted imaging of the brain at 7 T

Ventral and rostral regions of the brain are of emerging importance for the MRI characterization of early dementia, traumatic brain injury and epilepsy. Unfortunately, standard single-shot echo planar diffusion-weighted imaging of these regions at high fields is contaminated by severe imaging artifacts in the vicinity of air–tissue interfaces. To mitigate these artifacts and improve visualization of the temporal and frontal lobes at 7 T, we applied a reduced field-of-view strategy, enabled by outer volume suppression (OVS) with novel quadratic phase radiofrequency (RF) pulses, combined with partial Fourier and parallel imaging methods. The new acquisition greatly reduced the level of artifacts in six human subjects (including four patients with early symptoms of dementia).