Late Quaternary uplift and subsidence of the west coast of Tanna, south Vanuatu, southwest Pacific: U-Th ages of raised coral reefs in the Median Sedimentary Basin

Twelve Late Quaternary TIMS U-Th ages are reported here from 10 coral samples collected in situ from five transgressive coral/algal raised reefs (height: max. 113 m, min. 8 m) and two raised lagoonal deposits (height: max. 18 m, min. 8 m) along and near the west coast of Tanna, which lies in the Median Sedimentary Basin of South Vanuatu, southwest Pacific. These reefs and raised lagoonal deposits represent several age groups: (i) 215 ka (marine oxygen-isotope stage 7) penultimate interglacial (highest elevation and oldest); (ii) one lagoonal deposit of ca 127 ka (marine oxygen-isotope stage 5e); (iii) three last interglacial reefs with ages 102, 89 and 81 ka (representing marine oxygen-isotope stages 5c, 5b and 5a, respectively, of the latter part of the last interglacial); (iv) a lagoonal deposit with a 92 ka age (5b); and (v) a Holocene reef (age >5.7-5.0 ka) (lowest elevation and youngest). A ca 4.9 ka regressive reef (at elevation of 1.5 m above sea-level) is consistent with an island-wide 6.5 m uplift (probably largely coseismic), and a probable further island-wide uplift occurred in the late Holocene. The U-series ages taken together with the heights of transgressive reefs show that uplift since 215 ka was, on average, at similar to0.52 mm/y; although since 5 ka the uplift rate was, on average, similar to1.6 mm/y (the assumption being that a 1.5 m above sea-level reef has a coseismic origin). Elevation of transgressive reefs 5a, 5b and 5c and their ages indicates an island-wide subsidence during the period ?124-89 ka (i.e. Late Quaternary uplift/subsidence was jerky). Late Quaternary uplift/subsidence on the northwest coast of Tanna is considered to be due to irregular thicknesses of crust being subducted beneath Tanna.

Site-directed mutagenesis and kinetic studies of the West Nile virus NS3 protease identify key enzyme-substrate interactions

The flavivirus West Nile virus (WNV) has spread rapidly throughout the world in recent years causing fever, meningitis, encephalitis, and fatalities. Because the viral protease NS2B/NS3 is essential for replication, it is attracting attention as a potential therapeutic target, although there are currently no antiviral inhibitors for any flavivirus. This paper focuses on elucidating interactions between a hexapeptide substrate (Ae-KPGLKR-p-nitroanilide) and residues at S1 and S2 in the active site of WNV protease by comparing the catalytic activities of selected mutant recombinant proteases in vitro. Homology modeling enabled the predictions of key mutations in VWNV NS3 protease at S1 (V115A/F, D129A/ E/N, S135A, Y150A/F, S160A, and S163A) and S2 (N152A) that might influence substrate recognition and catalytic efficiency. Key conclusions are that the substrate P1 Arg strongly interacts with S1 residues Asp-129, Tyr-150, and Ser-163 and, to a lesser extent, Ser-160, and P2 Lys makes an essential interaction with Asn-152 at S2. The inferred substrate-enzyme interactions provide a basis for rational protease inhibitor design and optimization. High sequence conservation within flavivirus proteases means that this study may also be relevant to design of protease inhibitors for other flavivirus proteases.

How the west was once: vegetation change in south-west Queensland from 1930 to 1995

Conflicting perceptions of past and present rangeland condition and limited historical data have led to debate regarding the management of vegetation in pastoral landscapes both internationally and in Australia. In light of this controversy we have sought to provide empirical evidence to determine the trajectory of vegetational change in a semi-arid rangeland for a significant portion of the 20th century using a suite of proxy measures. Ambathala Station, approximately 780 km west of Brisbane, in the semi-arid rangelands of south-western Queensland, Australia. We excavated stratified deposits of sheep manure which had accumulated beneath a shearing shed between the years 1930 and 1995. Multi-proxy data, including pollen and leaf cuticle analyses and analysis of historical aerial photography were coupled with a fine resolution radiocarbon chronology to generate a near annual history of vegetation on the property and local area. Aerial photography indicates that minor (< 5%) increases in the density of woody vegetation took place between 1951 and 1994 in two thirds of the study area not subjected to clearing. Areas that were selectively or entirely cleared prior to the 1950s (approximately 16% of the study area) had recovered to almost 60% of their original cover by the 1994 photo period. This slight thickening is only partially evident from pollen and leaf cuticle analyses of sheep faeces. Very little change in vegetation is revealed over the nearly 65 years based on the relative abundances of pollen taxonomic groups. Microhistological examination of sheep faeces provides evidence of dramatic changes in sheep diet. The majority of dietary changes are associated with climatic events of sustained above-average rainfall or persistent drought. Most notable in the dietary analysis is the absence of grass during the first two decades of the record. In contrast to prevailing perceptions and limited research into long-term vegetation change in the semi-arid areas of eastern Australia, the record of vegetation change at the Ambathala shearing shed indicates only a minor increase in woody vegetation cover and no decrease in grass cover on the property over the 65 years of pastoral activity covered by the study. However, there are marked changes in the abundance of grass cuticles in sheep faeces. The appearance and persistence of grass in sheep diets from the late 1940s can be attributed to the effects of periods of high rainfall and possibly some clearing and thinning of vegetation. Lower stock numbers may have allowed grass to persist through later drought years. The relative abundances of major groups of plant pollen have not changed significantly over the past 65 years.

Regulated Cleavages at the West Nile Virus NS4A-2K-NS4B Junctions Play a Major Role in Rearranging Cytoplasmic Membranes and Golgi Trafficking of the NS4A Protein

A common feature associated with the replication of most RNA viruses is the formation of a unique membrane environment encapsulating the viral replication complex. For their part, flaviviruses are no exception, whereupon infection causes a dramatic rearrangement and induction of unique membrane structures within the cytoplasm of infected cells. These virus-induced membranes, termed paracrystalline arrays, convoluted membranes, and vesicle packets, all appear to have specific functions during replication and are derived from different organelles within the host cell. The aim of this study was to identify which protein(s) specified by the Australian strain of West Nile virus, Kunjin virus (KUNV), are responsible for the dramatic membrane alterations observed during infection. Thus, we have shown using immunolabeling of ultrathin cryosections of transfected cells that expression of the KUNV polyprotein intermediates NS4A-4B and NS213-34A, as well as that of individual NS4A proteins with and without the C-terminal transmembrane domain 2K, resulted in different degrees of rearrangement of cytoplasmic membranes. The formation of the membrane structures characteristic for virus infection required coexpression of an NS4A-NS4B cassette with the viral protease NS2B-3pro which was shown to be essential for the release of the individual NS4A and NS4B proteins. Individual expression of NS4A protein retaining the C-terminal transmembrane domain 2K resulted in the induction of membrane rearrangements most resembling virus-induced structures, while removal of the 2K domain led to a less profound membrane rearrangement but resulted in the redistribution of the NS4A protein to the Golgi apparatus. The results show that cleavage of the KUNV polyprotein NS4A-4B by the viral protease is the key initiation event in the induction of membrane rearrangement and that the NS4A protein intermediate containing the uncleaved C-terminal transmembrane domain plays an essential role in these membrane rearrangements.

Origin of the West Nile virus responsible for an outbreak of encephalitis in the northeastern United States

In Late summer 1999, an outbreak of human encephalitis occurred in the northeastern United States that was concurrent with extensive mortality in crows (Corvus species) as well as the deaths of several exotic birds at a zoological park in the same area. Complete genome sequencing of a flavivirus isolated from the brain of a dead Chilean flamingo (Phoenicopterus chilensis), together with partial sequence analysis of envelope glycoprotein (E-glycoprotein) genes amplified from several other species including mosquitoes and two fatal human cases, revealed that West Nile (WN) virus circulated in natural transmission cycles and was responsible for the human disease. Antigenic mapping with E-glycoprotein-specific monoclonal antibodies and E-glycoprotein phylogenetic analysis confirmed these viruses as WN. This North American WN virus was most closely related to a WN virus isolated from a dead goose in Israel in 1998.

NS1 protein secretion during the acute phase of West Nile virus infection

The West Nile virus (WNV) nonstructural protein NS1 is a protein of unknown function that is found within, associated with, and secreted from infected cells. We systematically investigated the kinetics of NS1 secretion in vitro and in vivo to determine the potential use of this protein as a diagnostic marker and to analyze NS1 secretion in relation to the infection cycle. A sensitive antigen capture enzyme-linked immunosorbent assay (ELISA) for detection of WNW NS1 (polyclonal-ACE) was developed, as well as a capture ELISA for the specific detection of NS1 multimers (4G4-ACE). The 4G4-ACE detected native NS1 antigens at high sensitivity, whereas the polyclonal-ACE had a higher specificity for recombinant forms of the protein. Applying these assays we found that only a small fraction of intracellular NS1 is secreted and that secretion of NS1 in tissue culture is delayed compared to the release of virus particles. In experimentally infected hamsters, NS1 was detected in the serum between days 3 and 8 postinfection, peaking on day 5, the day prior to the onset of clinical disease; immunoglobulin M (IgM) antibodies were detected at low levels on day 5 postinfection. Although real-time PCR gave the earliest indication of infection (day 1), the diagnostic performance of the 4G4-ACE was comparable to that of real-time PCR during the time period when NS1 was secreted. Moreover, the 4G4-ACE was found to be superior in performance to both the IgM and plaque assays during this time period, suggesting that NS1 is a viable early diagnostic marker of WNV infection.