Conserved functional domains and a novel tertiary interaction near the pseudoknot drive translational activity of hepatitis C virus and hepatitis C virus-like internal ribosome entry sites

The translational activity of the hepatitis C virus (HCV) internal ribosome entry site (IRES) and other HCV-like IRES RNAs depends on structured RNA elements in domains II and III, which serve to recruit the ribosomal 40S subunit, eukaryotic initiation factor (eIF) 3 and the ternary eIF2/Met-tRNAiMet/GTP complex and subsequently domain II assists subunit joining. Porcine teschovirus-1 talfan (PTV-1) is a member of the Picornaviridae family, with a predicted HCV-like secondary structure, but only stem-loops IIId and IIIe in the 40S-binding domain display significant sequence conservation with the HCV IRES. Here, we use chemical probing to show that interaction sites with the 40S subunit and eIF3 are conserved between HCV and HCV-like IRESs. In addition, we reveal the functional role of a strictly conserved co-variation between a purine–purine mismatch near the pseudoknot (A–A/G) and the loop sequence of domain IIIe (GAU/CA). These nucleotides are involved in a tertiary interaction, which serves to stabilize the pseudoknot structure and correlates with translational efficiency in both the PTV-1 and HCV IRES. Our data demonstrate conservation of functional domains in HCV and HCV-like IRESs including a more complex structure surrounding the pseudoknot than previously assumed

Spectroscopic examinations of hydro- and glaciovolcanic basaltic tuffs: Modes of alteration and relevance for Mars

Hydro- and glaciovolcanism are processes that have taken place on both Earth and Mars. The amount of materials produced by these processes that are present in the martian surface layer is unknown, but may be substantial. We have used Mars rover analogue analysis techniques to examine altered tuff samples collected from multiple hydrovolcanic features, tuff rings and tuff cones, in the American west and from glaciovolcanic hyaloclastite ridges in Washington state and in Iceland. Analysis methods include VNIR-SWIR reflectance, MWIR thermal emissivity, thin section petrography, XRD, XRF, and Mössbauer spectroscopy. We distinguish three main types of tuff that differ prominently in petrography and VNIR-SWIR reflectance: minimally altered sideromelane tuff, gray to brown colored smectite-bearing tuff, and highly palagonitized tuff. Differences are also observed between the tuffs associated with hydrovolcanic tuff rings and tuff cones and those forming glaciovolcanic hyaloclastite ridges. For the locations sampled, hydrovolcanic palagonite tuffs are more smectite and zeolite rich while the palagonitized hyaloclastites from the sampled sites are largely devoid of zeolites and relatively lacking in smectites as well. The gray to brown colored tuffs are only observed in the hydrovolcanic deposits and appear to represent a distinct alteration pathway, with formation of smectites without associated palagonite formation. This is attributed to lower temperatures and possibly longer time scale alteration. Altered hydro- or glaciovolcanic materials might be recognized on the surface of Mars with rover-based instrumentation based on the results of this study

The Acid Test of Fluoride: How pH Modulates Toxicity

Background: It is not known why the ameloblasts responsible for dental enamel formation are uniquely sensitive to fluoride ($F^−$). Herein, we present a novel theory with supporting data to show that the low pH environment of maturating stage ameloblasts enhances their sensitivity to a given dose of $F^−$. Enamel formation is initiated in a neutral pH environment (secretory stage); however, the pH can fall to below 6.0 as most of the mineral precipitates (maturation stage). Low pH can facilitate entry of $F^−$ into cells. Here, we asked if $F^−$ was more toxic at low pH, as measured by increased cell stress and decreased cell function. Methodology/Principal Findings: Treatment of ameloblast-derived LS8 cells with $F^−$ at low pH reduced the threshold dose of $F^−$ required to phosphorylate stress-related proteins, PERK, eIF2α, JNK and c-jun. To assess protein secretion, LS8 cells were stably transduced with a secreted reporter, Gaussia luciferase, and secretion was quantified as a function of $F^−$ dose and pH. Luciferase secretion significantly decreased within 2 hr of $F^−$ treatment at low pH versus neutral pH, indicating increased functional toxicity. Rats given 100 ppm $F^−$ in their drinking water exhibited increased stress-mediated phosphorylation of eIF2α in maturation stage ameloblasts (pH<6.0) as compared to secretory stage ameloblasts (pH∼7.2). Intriguingly, $F^−$-treated rats demonstrated a striking decrease in transcripts expressed during the maturation stage of enamel development (Klk4 and Amtn). In contrast, the expression of secretory stage genes, AmelX, Ambn, Enam and Mmp20, was unaffected. Conclusions: The low pH environment of maturation stage ameloblasts facilitates the uptake of $F^−$, causing increased cell stress that compromises ameloblast function, resulting in dental fluorosis

"Our method" for teaching practical French.

Mode of access: Internet.MAIN; 984z.G589: Letter inserted.MAIN; 984z.G589: Autographed presentation copy

Personal Papers (MS 80-0002)

Telegram sent by Leon Godchaux to Otto Marx and Isaac H. Kempner Jr. discussing hotel room reservation details

Testing geopressured geothermal reservoirs in existing wells: Detailed completion prognosis for geopressured-geothermal well of opportunity, prospect #7

This book is a detailed prognosis covering the acquisition, completion, drilling, testing and abandonment of the Frank A. Godchaux, III, Well No. 1 under the Wells of Opportunity Program. The well is located approximately 12 miles southeast of the city of Abbeville, Louisiana. Eaton Operating Company proposes to test a section of the Planulina sand at a depth ranging from 15,584 to 15,692 feet. The reservoir pressure is estimated to be 14,480 psi and the temperature of the formation water is expected to be 298 F. The water salinity is calculated to be 75,000 ppm. The well is expected to produce 20,000 barrels of water per day with a gas content of 44 standard cubic feet pre barrel. The well was acquired from C and K Petroleu, Inc. on March 20, 1981. C and K abandoned the well at a total depth of 16,000 feet. The well has a 7-5/8 inches liner set at 13,387 feet. Eaton proposes to set 5-1/2 inch casing at 16,000 feet and produce the well through the casing using a 2-3/8 inch tubing string for wireline protection and for pressure control. A 4,600 foot saltwater disposal well will be drilled on the site and testing will be conducted similar to previous Eaton tests. The total estimated cost to perform the work is $2,959,000. An optional test from 14,905 to 15,006 feet may be performed after the original test and will require a workover with a rig on location to perform the plugback. The surface production equipment utilized on previous Eaton WOO tests will be utilized on this test. This equipment has worked satisfactorily and all parties involved in the testing are familiar with its operation. The Institute of Gas Technology and Mr. Don Clark will handle the sampling and testing and reservoir evaluation, respectively, as on the previous Eaton tests