Risky business Doern, G. Bruce; Reed, Ted
Risky business,
c2000, 20000905, 2000, 2000-01-01
eBook
The essays in this volume ask what risks Canadians might be exposed to as fiscal pressures strain the capacity of regulators in areas such as food, drugs, pesticides, fisheries, and the environment.
Antibodies and antibody-derived macromolecules have established themselves as the mainstay in protein-based therapeutic molecules (biologics). Our knowledge of the structure-function relationships of ...antibodies provides a platform for protein engineering that has been exploited to generate a wide range of biologics for a host of therapeutic indications. In this review, our basic understanding of the antibody structure is described along with how that knowledge has leveraged the engineering of antibody and antibody-related therapeutics having the appropriate antigen affinity, effector function, and biophysical properties. The platforms examined include the development of antibodies, antibody fragments, bispecific antibody, and antibody fusion products, whose efficacy and manufacturability can be improved via humanization, affinity modulation, and stability enhancement. We also review the design and selection of binding arms, and avidity modulation. Different strategies of preparing bispecific and multispecific molecules for an array of therapeutic applications are included.
We describe a new class of propagation-invariant light beams with Fourier transform given by an eigenfunction of the quantum mechanical pendulum. These beams, whose spectra (restricted to a circle) ...are doubly-periodic Mathieu functions in azimuth, depend on a field strength parameter. When the parameter is zero, pendulum beams are Bessel beams, and as the parameter approaches infinity, they resemble transversely propagating one-dimensional Gaussian wavepackets (Gaussian beam-beams). Pendulum beams are the eigenfunctions of an operator which interpolates between the squared angular momentum operator and the linear momentum operator. The analysis reveals connections with Mathieu beams, and insight into the paraxial approximation.
The major genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) is a C9orf72 G
C
repeat expansion
. Proposed mechanisms by which the expansion causes c9FTD/ALS ...include toxicity from repeat-containing RNA and from dipeptide repeat proteins translated from these transcripts. To investigate the contribution of poly(GR) dipeptide repeat proteins to c9FTD/ALS pathogenesis in a mammalian in vivo model, we generated mice that expressed GFP-(GR)
in the brain. GFP-(GR)
mice developed age-dependent neurodegeneration, brain atrophy, and motor and memory deficits through the accumulation of diffuse, cytoplasmic poly(GR). Poly(GR) co-localized with ribosomal subunits and the translation initiation factor eIF3η in GFP-(GR)
mice and, of importance, in c9FTD/ALS patients. Combined with the differential expression of ribosome-associated genes in GFP-(GR)
mice, these findings demonstrate poly(GR)-mediated ribosomal distress. Indeed, poly(GR) inhibited canonical and non-canonical protein translation in HEK293T cells, and also induced the formation of stress granules and delayed their disassembly. These data suggest that poly(GR) contributes to c9FTD/ALS by impairing protein translation and stress granule dynamics, consequently causing chronic cellular stress and preventing cells from mounting an effective stress response. Decreasing poly(GR) and/or interrupting interactions between poly(GR) and ribosomal and stress granule-associated proteins may thus represent potential therapeutic strategies to restore homeostasis.
Enzymatic formation of glutamate is critical to numerous biological pathways. However, current methods for assaying the activities of glutamate-forming enzymes are not particularly suitable for ...high-throughput screening in drug discovery. We present a continuous-read, fluorometric assay for high-throughput analysis of glutaminases. This assay is adapted to a microplate format and employs glutamate oxidase and horseradish peroxidase to couple glutamate formation to production of the fluorescent reporter molecule, resorufin, for enhancement of sensitivity (M. Zhou, Z. Diwu, N. Panchuk-Voloshina, and R. P. Haughland, 1997, Anal. Biochem. 253, 162–168). Described herein is the selection of suitable levels of coupling enzymes for optimal kinetic response and lag time of the reporter system, based on the kinetic characteristics of the individual coupling enzymes. Finally, implementation of the assay in a format for high-throughput kinetic analysis of glutaminases is demonstrated for Escherichia coli carbamoyl phosphate synthase. Derived kinetic constants are comparable to literature values determined using a variety of assay techniques.
The magnitudes of beam shifts (Goos-H\"anchen and Imbert-Fedorov, spatial and angular) are greatly enhanced when a reflected light beam is postselected by an analyzer, by analogy with superweak ...measurements in quantum theory. Particularly strong enhancements can be expected close to angles at which no light is transmitted for a fixed initial and final polarizations. We derive a formula for the angular and spatial shifts at such angles (which includes the Brewster angle), and we show that their maximum size is limited by higher-order terms from the reflection coefficients occurring in the Artmann shift formula.
The HIV-1 fusion peptide, comprising 15 to 20 hydrophobic residues at the N terminus of the Env-gp41 subunit, is a critical component of the virus-cell entry machinery. Here, we report the ...identification of a neutralizing antibody, N123-VRC34.01, which targets the fusion peptide and blocks viral entry by inhibiting conformational changes in gp120 and gp41 subunits of Env required for entry. Crystal structures of N123-VRC34.01 liganded to the fusion peptide, and to the full Env trimer, revealed an epitope consisting of the N-terminal eight residues of the gp41 fusion peptide and glycan N88 of gp120, and molecular dynamics showed that the N-terminal portion of the fusion peptide can be solvent-exposed. These results reveal the fusion peptide to be a neutralizing antibody epitope and thus a target for vaccine design.
A set of pupil apodization functions for use with a vortex coronagraph on telescopes with obscured apertures is presented. We show analytically that pupil amplitudes given by real-valued Zernike ...polynomials offer ideal on-axis starlight cancellation when applied to unobscured circular apertures. The charge of the vortex phase element must be a nonzero even integer, greater than the sum of the degree and the absolute value of its azimuthal order of the Zernike polynomial. Zero-valued lines and points of Zernike polynomials, or linear combinations thereof, can be matched to obstructions in the pupils of ground-based telescopes to improve the contrast achieved by a vortex coronagraph. This approach works well in the presence of a central obscuration and radial support structures. We analyze the contrast, off-axis throughput, and post-coronagraph point spread functions of an apodized vortex coronagraph designed for the European Extremely Large Telescope (E-ELT). This technique offers very good performance on apertures with large obscuring support structures similar to those on future 30-40m class ground-based telescopes.
Following Nye and Berry's analogy with crystal dislocations, an approach to the Burgers vector of a wave dislocation (phase singularity, optical vortex) is proposed. It is defined to be a regularized ...phase gradient evaluated at the phase singularity, and is computed explicitly. The screw component of this vector is naturally related to the helicoidal twisting of wavefronts along a vortex line, and is related to the helicity of the phase gradient. The edge component is related to the nearby current flow (defined by the phase gradient) perpendicular to the vortex, and the distribution of this component is found numerically for random two-dimensional monochromatic waves.
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