Raman injection laser

Stimulated Raman scattering is a nonlinear optical process that, in a broad variety of materials, enables the generation of optical gain at a frequency that is shifted from that of the incident radiation by an amount corresponding to the frequency of an internal oscillation of the material. This effect is the basis for a broad class of tunable sources known as Raman lasers. In general, these sources have only small gain (approximately 10(-9) cm W(-1)) and therefore require external pumping with powerful lasers, which limits their applications. Here we report the realization of a semiconductor injection Raman laser designed to circumvent these limitations. The physics underlying our device differs in a fundamental way from existing Raman lasers: it is based on triply resonant stimulated Raman scattering between quantum-confined states within the active region of a quantum cascade laser that serves as an internal optical pump--the device is driven electrically and no external laser pump is required. This leads to an enhancement of orders of magnitude in the Raman gain, high conversion efficiency and low threshold. Our lasers combine the advantages of nonlinear optical devices and of semiconductor injection lasers, and could lead to a new class of compact and wavelength-agile mid-and far-infrared light sources.     

Rapid,quantitative adipose conversion of chicken fibroblasts by high concentrations of chicken serum or plasma

Summary Rapid, quantitative adipose conversion of chicken fibroblasts occurs when these cells are cultured in undiluted commercial chicken serum or plasma. Fresh serum and plasma acquire this property after ageing.These studies were carried out with grants from the Richardson Foundation, the National Cancer Institute of Canada, and the Canadian Heart Foundations.To whom requests for reprints should be sent.     

Sequence-based characterization of structural variation in the mouse genome

Structural variation is widespread in mammalian genomes and is an important cause of disease, but just how abundant and important structural variants (SVs) are in shaping phenotypic variation remains unclear. Without knowing how many SVs there are, and how they arise, it is difficult to discover what they do. Combining experimental with automated analyses, we identified 711,920 SVs at 281,243 sites in the genomes of thirteen classical and four wild-derived inbred mouse strains. The majority of SVs are less than 1?kilobase in size and 98% are deletions or insertions. The breakpoints of 160,000 SVs were mapped to base pair resolution, allowing us to infer that insertion of retrotransposons causes more than half of SVs. Yet, despite their prevalence, SVs are less likely than other sequence variants to cause gene expression or quantitative phenotypic variation. We identified 24 SVs that disrupt coding exons, acting as rare variants of large effect on gene function. One-third of the genes so affected have immunological functions.     

Improved molecular replacement by density- and energy-guided protein structure optimization

Molecular replacement procedures, which search for placements of a starting model within the crystallographic unit cell that best account for the measured diffraction amplitudes, followed by automatic chain tracing methods, have allowed the rapid solution of large numbers of protein crystal structures. Despite extensive work, molecular replacement or the subsequent rebuilding usually fail with more divergent starting models based on remote homologues with less than 30% sequence identity. Here we show that this limitation can be substantially reduced by combining algorithms for protein structure modelling with those developed for crystallographic structure determination. An approach integrating Rosetta structure modelling with Autobuild chain tracing yielded high-resolution structures for 8 of 13 X-ray diffraction data sets that could not be solved in the laboratories of expert crystallographers and that remained unsolved after application of an extensive array of alternative approaches. We estimate that the new method should allow rapid structure determination without experimental phase information for over half the cases where current methods fail, given diffraction data sets of better than 3.2?? resolution, four or fewer copies in the asymmetric unit, and the availability of structures of homologous proteins with >20% sequence identity.     

Visualization Methodology for Multidimensional Scaling

These uncertainties will be addressed by the following interactive techniques: (a) algorithm animation, random restarts, and manual editing of configurations, (b) interactive control over parameters that determine the criterion and its minimization, (c) diagnostics for pinning down artifactual point configurations, and (d) restricting MDS to subsets of objects and subsets of pairs of objects. A system, called "XGvis", which implments these techniques, is freely available with the "XGobi" distribution. XGobi is a multivariate data visualization system that is used here for visualizing point configurations.     

Restoration of type VII collagen expression and function in dystrophic epidermolysis bullosa

Dystrophic epidermolysis bullosa (DEB) is a family of inherited mechano-bullous disorders caused by mutations in the human type VII collagen gene (COL7A1). Individuals with DEB lack type VII collagen and anchoring fibrils, structures that attach epidermis and dermis. The current lack of treatment for DEB is an impetus to develop gene therapy strategies that efficiently transfer and stably express genes delivered to skin cells in vivo. In this study, we delivered and expressed full-length type VII collagen using a self-inactivating minimal lentivirus-based vector. Transduction of lentiviral vectors containing the COL7A1 transgene into recessive DEB (RDEB) keratinocytes and fibroblasts (in which type VII collagen was absent) resulted in persistent synthesis and secretion of type VII collagen. Unlike RDEB parent cells, the gene-corrected cells had normal morphology, proliferative potential, matrix attachment and motility. We used these gene-corrected cells to regenerate human skin on immune-deficient mice. Human skin regenerated by gene-corrected RDEB cells had restored expression of type VII collagen and formation of anchoring fibrils at the dermal-epidermal junction in vivo. These studies demonstrate that it is possible to restore type VII collagen gene expression in RDEB skin in vivo.