An improved rhodopsin/EGFP fusion protein for use in the generation of transgenic Xenopus laevis

AbstractPrevious studies by Papermaster and coworkers introduced the use of rhodopsin–green fluorescent protein (rho–GFP) fusion proteins in the construction of transgenic Xenopus laevis with retinal rod photoreceptor cell-specific transgene expression [Moritz et al., J. Biol. Chem. 276 (2001) 28242–28251]. These pioneering studies have helped to develop the Xenopus system not only for use in the investigation of rhodopsin biosynthesis and targeting, but for studies of the phototransduction cascade as well. However, the rho–GFP fusion protein used in the earlier work had only 50% of the specific activity of wild-type rhodopsin for activation of transducin and only 10% of the activity of wild-type in rhodopsin kinase assays. While not a problem for the biosynthesis studies, this does present a problem for investigation of the phototransduction cascade. We report here an improved rhodopsin/EGFP fusion protein in which placement of the EGFP domain at the C-terminus of rhodopsin results in wild-type activity for activation of transducin, wild-type ability to serve as a substrate for rhodopsin kinase, and wild-type localization of the protein to the rod photoreceptor cell outer segment in transgenic X. laevis

Data Recovery Investigations: Murvaul Creek Site (41PN175), Panola County, Texas

This report summarizes the archeological findings of the 2011 data recovery investigations at the Murvaul Creek site, 41PN175, in far northeastern Texas in Panola County. The site is located along Farm-to-Market Road (FM) 10 approximately 1 mile north of Gary, Texas (Figure 1). Geo-Marine, Inc. (GMI), performed this work under contract to the Texas Department of Transportation, Environmental Affairs Division (TxDOT ENV) under the Texas Antiquities Permit Number 5879 (Work Authorization [WA] 579 06 SA005; WA 590 08 SA005; CSJ:1222-01-014; Geo-Marine project numbers 22005.00.06 and 22005.00.09). The fieldwork for this project was conducted in advance of the planned widening of FM 10 that was to replace three bridges and a culvert over Murvaul Creek with a larger structure and shift the road approximately 26 meters (m; 85 feet [ft]) to the east. Since the planned improvements of FM 10 would result in the loss of information at the Murvaul Creek site—a site that was recommended eligible for inclusion in the National Register of Historic Places (NRHP) and for designation as a State Antiquities Landmark (SAL; formerly State Archeological Landmark)—the current data recovery investigations were initiated. The data recovery investigations were conducted between February 7, 2011, and April 3, 2011. During this period, the fieldwork was conducted in several stages: site clearing, geophysical survey, 50-x-50-centimeter (cm) excavations, block excavations, and mechanical site scraping. With the exception of the site clearing stage, the results of each of the fieldwork stages are reviewed individually in this report. The investigations resulted in the documentation of numerous features that appeared to have been the remains of a small Middle-to-Late Caddo settlement or farmstead situated on the edge of an interfluve south of the Murvaul Creek floodplain. Additionally, materials pertaining to the Archaic period were documented across the site. Although the site has been intensively studied within the TxDOT right-of-way (ROW), both the current investigations and previous work were limited to the ROW (cf. Cliff and Perttula 2002). Hence, the site is very likely larger than has been adequately documented

In vivo imaging of extracellular matrix remodeling by tumor-associated fibroblasts.

Here we integrated multiphoton laser scanning microscopy and the registration of second harmonic generation images of collagen fibers to overcome difficulties in tracking stromal cell-matrix interactions for several days in live mice. We show that the matrix-modifying hormone relaxin increased tumor-associated fibroblast (TAF) interaction with collagen fibers by stimulating beta1-integrin activity, which is necessary for fiber remodeling by matrix metalloproteinases

Enamelin Is Critical for Ameloblast Integrity and Enamel Ultrastructure Formation

Mutations in the human enamelin gene cause autosomal dominant hypoplastic amelogenesis imperfecta in which the affected enamel is thin or absent. Study of enamelin knockout NLS-lacZ knockin mice revealed that mineralization along the distal membrane of ameloblast is deficient, resulting in no true enamel formation. To determine the function of enamelin during enamel formation, we characterized the developing teeth of the Enam−/− mice, generated amelogenin-driven enamelin transgenic mouse models, and then introduced enamelin transgenes into the Enam−/− mice to rescue enamel defects. Mice at specific stages of development were subjected to morphologic and structural analysis using β-galactosidase staining, immunohistochemistry, and transmission and scanning electron microscopy. Enamelin expression was ameloblast-specific. In the absence of enamelin, ameloblasts pathology became evident at the onset of the secretory stage. Although the aggregated ameloblasts generated matrix-containing amelogenin, they were not able to create a well-defined enamel space or produce normal enamel crystals. When enamelin is present at half of the normal quantity, enamel was thinner with enamel rods not as tightly arranged as in wild type suggesting that a specific quantity of enamelin is critical for normal enamel formation. Enamelin dosage effect was further demonstrated in transgenic mouse lines over expressing enamelin. Introducing enamelin transgene at various expression levels into the Enam−/− background did not fully recover enamel formation while a medium expresser in the Enam+/− background did. Too much or too little enamelin abolishes the production of enamel crystals and prism structure. Enamelin is essential for ameloblast integrity and enamel formation

Enamelin is critical for ameloblast integrity and enamel ultrastructure formation

Mutations in the human enamelin gene cause autosomal dominant hypoplastic amelogenesis imperfecta in which the affected enamel is thin or absent. Study of enamelin knockout NLS-lacZ knockin mice revealed that mineralization along the distal membrane of ameloblast is deficient, resulting in no true enamel formation. To determine the function of enamelin during enamel formation, we characterized the developing teeth of the Enam-/- mice, generated amelogenin-driven enamelin transgenic mouse models, and then introduced enamelin transgenes into the Enam-/- mice to rescue enamel defects. Mice at specific stages of development were subjected to morphologic and structural analysis using β-galactosidase staining, immunohistochemistry, and transmission and scanning electron microscopy. Enamelin expression was ameloblast-specific. In the absence of enamelin, ameloblasts pathology became evident at the onset of the secretory stage. Although the aggregated ameloblasts generated matrix-containing amelogenin, they were not able to create a well-defined enamel space or produce normal enamel crystals. When enamelin is present at half of the normal quantity, enamel was thinner with enamel rods not as tightly arranged as in wild type suggesting that a specific quantity of enamelin is critical for normal enamel formation. Enamelin dosage effect was further demonstrated in transgenic mouse lines over expressing enamelin. Introducing enamelin transgene at various expression levels into the Enam -/- background did not fully recover enamel formation while a medium expresser in the Enam+/- background did. Too much or too little enamelin abolishes the production of enamel crystals and prism structure. Enamelin is essential for ameloblast integrity and enamel formation. © 2014 Hu et al

The Dependence of Star Formation History and Internal Structure on Stellar Mass for 10^5 Low-Redshift Galaxies

We study the relations between stellar mass, star formation history, size and internal structure for a complete sample of 122,808 galaxies drawn from the Sloan Digital Sky Survey. We show that low-redshift galaxies divide into two distinct families at a stellar mass of 3 \times 10^10 M_sol. Lower mass galaxies have young stellar populations, low surface mass densities and the low concentrations typical of disks. A significant fraction of the lowest mass galaxies in our sample have experienced recent starbursts. At given stellar mass, the sizes of low mass galaxies are log- normally distributed with dispersion sigma(ln R_50) \sim 0.5, in excellent agreement with the idea that they form with little angular momentum loss through cooling and condensation in a gravitationally dominant dark matter halo. Their median stellar surface mass density scales with stellar mass as mu* propto M_*^0.54, suggesting that the stellar mass of a disk galaxy is proprtional to the three halves power of its halo mass. This suggests that the efficiency of the conversion of baryons into stars in low mass galaxies increases in propor- tion to halo mass, perhaps as a result of supernova feedback processes. At stellar masses above 3 \times 10^10 M_sol, there is a rapidly increasing frac- tion of galaxies with old stellar populations, high surface mass densities and high concentrations typical of bulges. In this regime, the size distribution is log-normal, but its dispersion decreases rapidly with increasing stellar mass and the median mass surface density is approximately constant. This suggests that the star formation efficiency decreases in the highest mass halos, and that little star formation occurs in massive galaxies once they have assembled.Comment: accepted by MNRAS, some changes to results as a result of improvements in stellar mass estimates as decribed in Paper

The Long-Baseline Neutrino Experiment: Exploring Fundamental Symmetries of the Universe

The preponderance of matter over antimatter in the early Universe, the dynamics of the supernova bursts that produced the heavy elements necessary for life and whether protons eventually decay --- these mysteries at the forefront of particle physics and astrophysics are key to understanding the early evolution of our Universe, its current state and its eventual fate. The Long-Baseline Neutrino Experiment (LBNE) represents an extensively developed plan for a world-class experiment dedicated to addressing these questions. LBNE is conceived around three central components: (1) a new, high-intensity neutrino source generated from a megawatt-class proton accelerator at Fermi National Accelerator Laboratory, (2) a near neutrino detector just downstream of the source, and (3) a massive liquid argon time-projection chamber deployed as a far detector deep underground at the Sanford Underground Research Facility. This facility, located at the site of the former Homestake Mine in Lead, South Dakota, is approximately 1,300 km from the neutrino source at Fermilab -- a distance (baseline) that delivers optimal sensitivity to neutrino charge-parity symmetry violation and mass ordering effects. This ambitious yet cost-effective design incorporates scalability and flexibility and can accommodate a variety of upgrades and contributions. With its exceptional combination of experimental configuration, technical capabilities, and potential for transformative discoveries, LBNE promises to be a vital facility for the field of particle physics worldwide, providing physicists from around the globe with opportunities to collaborate in a twenty to thirty year program of exciting science. In this document we provide a comprehensive overview of LBNE's scientific objectives, its place in the landscape of neutrino physics worldwide, the technologies it will incorporate and the capabilities it will possess.Comment: Major update of previous version. This is the reference document for LBNE science program and current status. Chapters 1, 3, and 9 provide a comprehensive overview of LBNE's scientific objectives, its place in the landscape of neutrino physics worldwide, the technologies it will incorporate and the capabilities it will possess. 288 pages, 116 figure

Effects of antiplatelet therapy on stroke risk by brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases: subgroup analyses of the RESTART randomised, open-label trial

Background Findings from the RESTART trial suggest that starting antiplatelet therapy might reduce the risk of recurrent symptomatic intracerebral haemorrhage compared with avoiding antiplatelet therapy. Brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases (such as cerebral microbleeds) are associated with greater risks of recurrent intracerebral haemorrhage. We did subgroup analyses of the RESTART trial to explore whether these brain imaging features modify the effects of antiplatelet therapy