Internet Legal Research Program Materials

Internet Legal Research presentations include: Google and Beyond: Finding Information Using Search Engines, and Evaluating Your Results; Why Pay For It Twice? How to Access Federal Materials in the Public Domaind; All Politics are Local: State and Local Resources; L is for Lawyer: An Alphabet of Handy Web Pages; Internet Basics: The Who, What, When, Where, Why and How of Internet Research for Lawyers; But Can I Get it in English? Finding Foreign Law; Blawgs, Podcasts, Wikis? Deciphering the Lingo and Evaluating Current Awareness Tool

Persistent Transport Barrier on the West Florida Shelf

Analysis of drifter trajectories in the Gulf of Mexico has revealed the existence of a region on the southern portion of the West Florida Shelf (WFS) that is not visited by drifters that are released outside of the region. This so-called ``forbidden zone'' (FZ) suggests the existence of a persistent cross-shelf transport barrier on the southern portion of the WFS. In this letter a year-long record of surface currents produced by a Hybrid-Coordinate Ocean Model simulation of the WFS is used to identify Lagrangian coherent structures (LCSs), which reveal the presence of a robust and persistent cross-shelf transport barrier in approximately the same location as the boundary of the FZ. The location of the cross-shelf transport barrier undergoes a seasonal oscillation, being closer to the coast in the summer than in the winter. A month-long record of surface currents inferred from high-frequency (HF) radar measurements in a roughly 60 km $\times$ 80 km region on the WFS off Tampa Bay is also used to identify LCSs, which reveal the presence of robust transient transport barriers. While the HF-radar-derived transport barriers cannot be unambiguously linked to the boundary of the FZ, this analysis does demonstrate the feasibility of monitoring transport barriers on the WFS using a HF-radar-based measurement system. The implications of a persistent cross-shelf transport barrier on the WFS for the development of harmful algal blooms on the shoreward side of the barrier are considered.Comment: Submitted to Geophysical Research Letter

Bicistronic Lentiviruses Containing a Viral 2A Cleavage Sequence Reliably Co-Express Two Proteins and Restore Vision to an Animal Model of LCA1

The disease processes underlying inherited retinal disease are complex and are not completely understood. Many of the corrective gene therapies designed to treat diseases linked to mutations in genes specifically expressed in photoreceptor cells restore function to these cells but fail to stop progression of the disease. There is growing consensus that effective treatments for these diseases will require delivery of multiple therapeutic proteins that will be selected to treat specific aspects of the disease process. The purpose of this study was to design a lentiviral transgene that reliably expresses all of the proteins it encodes and does so in a consistent manner among infected cells. We show, using both in vitro and in vivo analyses, that bicistronic lentiviral transgenes encoding two fluorescent proteins fused to a viral 2A-like cleavage peptide meet these expression criteria. To determine if this transgene design is suitable for therapeutic applications, we replaced one of the fluorescent protein genes with the gene encoding guanylate cyclase -1 (GC1) and delivered lentivirus carrying this transgene to the retinas of the GUCY1*B avian model of Leber congenital amaurosis – 1 (LCA1). GUCY1*B chickens carry a null mutation in the GC1 gene that disrupts photoreceptor function and causes blindness at hatching, a phenotype that closely matches that observed in humans with LCA1. We found that treatment of these animals with the 2A lentivector encoding GC1 restored vision to these animals as evidenced by the presence of optokinetic reflexes. We conclude that 2A-like peptides, with proper optimization, can be successfully incorporated into therapeutic vectors designed to deliver multiple proteins to neural retinal. These results highlight the potential of this vector design to serve as a platform for the development of combination therapies designed to enhance or prolong the benefits of corrective gene therapies

Formation of Multilayered Photosynthetic Biofilms in an Alkaline Thermal Spring in Yellowstone National Park, Wyoming▿

In this study, glass rods suspended at the air-water interface in the runoff channel of Fairy Geyser, Yellowstone National Park, WY, were used as a substratum to promote the development of biofilms that resembled multilayered mat communities in the splash zone at the geyser's source. This approach enabled the establishment of the temporal relationship between the appearance of Cyanobacteria, which ultimately formed the outer green layer, and the development of a red underlayer containing Roseiflexus-like Chloroflexi. This is the first study to define time-dependent successional events involved in the development of differently colored layers within microbial mats associated with many thermal features in Yellowstone National Park. Initial (1-month) biofilms were localized below the air-water interface (60 to 70°C), and the majority of retrieved bacterial sequence types were similar to Synechococcus and Thermus isolates. Biofilms then shifted, becoming established at and above the air-water interface after 3 months. During winter sampling (6 to 8 months), distinct reddish orange microcolonies were observed, consistent with the appearance of Roseiflexus-like sequences and bacteriochlorophyll a pigment signatures. Additionally, populations of Cyanobacteria diversified to include both unicellular and filamentous cell and sequence types. Distinct green and red layers were observed at 13 months. Planctomycetes-like sequences were also retrieved in high abundance from final biofilm layers and winter samples. Finally, biomass associated with geyser vent water contained Roseiflexus-like sequence types, in addition to other high-abundance sequence types retrieved from biofilm samples, supporting the idea that geothermal water serves as an inoculum for these habitats