Gag-Pol supplied in trans is efficiently packaged and supports viral function in human immunodeficiency virus type 1

The intracellular trafficking and subsequent incorporation of Gag-Pol into human immunodeficiency virus type 1 (HIV-1) remains poorly defined. Gag-Pol is encoded by the same mRNA as Gag and is generated by ribosomal frameshifting. The multimerization of Gag and Gag-Pol is an essential step in the formation of infectious viral particles. In this study, we examined whether the interaction between Gag and Gag-Pol is initiated during protein translation in order to facilitate the trafficking and subsequent packaging of Gag-Pol into the virion. A conditional cotransfection system was developed in which virion formation required the coexpression of two HIV-1-based plasmids, one that produces both Gag and Gag-Pol and one that only produces Gag-Pol. The Gag-Pol proteins were either immunotagged with a His epitope or functionally tagged with a mutation (K65R) in reverse transcriptase that is associated with drug resistance. Gag-Pol packaging was assessed to determine whether the Gag-Pol incorporated into the virion was preferentially packaged from the plasmid that expressed both Gag and Gag-Pol or whether it could be packaged from either plasmid. Our data show that translation of Gag and Gag-Pol from the same mRNA is not critical for virion packaging of the Gag-Pol polyprotein or for viral function.

Nodulated N-2-fixing Casuarina cunninghamiana is the sink for net N transfer from non-N-2-fixing Eucalyptus maculata via an ectomycorrhizal fungus Pisolithus sp using (NH4+)-N-15 or (NO3-)-N-15 supplied as ammonium nitrate

To determine the effects of nitrogen source on rates of net N transfer between plants connected by a common mycorrhizal network, we measured transfer of N supplied as (NH4NO3)-N-15-N-14 or (NH4NO3)-N-14-N-15 in three Casuarina/Eucalyptus treatments interconnected by a Pisolithus sp. The treatments were nonnodulated nonmycorrhizal/nonmycorrhizal; nonnodulated mycorrhizal/mycorrhizal; and nodulated mycorrhizal/mycorrhizal. Mycorrhization was 67% in Eucalyptus and 36% in Casuarina. N-2 fixation supplied 38% of the N in Casuarina. Biomass, N and N-15 contents were lowest in nonmycorrhizal plants and greatest in plants in the nodulated/mycorrhizal treatment. Nitrogen transfer was enhanced by mycorrhization and by nodulation, and was greater when N was supplied as (NH4+)-N-15 than (NO3-)-N-15. Nitrogen transfer rates were lowest in the nonmycorrhizal treatment for either N-15 source, and greatest in the nodulated, mycorrhizal treatment. Transfer was greater to Casuarina than to Eucalyptus and where ammonium rather than nitrate was the N source. Irrespective of N-15 source and of whether Casuarina or Eucalyptus was the N sink, net N transfer was low and was similar in both nonnodulated treatments. However, when Casuarina was the N sink in the nodulated, mycorrhizal treatment, net N transfer was much greater with (NH4+)-N-15 than with (NO3-)-N-15. High N demand by Casuarina resulted in greater net N transfer from the less N-demanding Eucalyptus. Net transfer of N from a non-N-2-fixing to an N-2-fixing plant may reflect the very high N demand of N-2-fixing species.

Effect of Mn deficiency and legume inoculation on rhizosphere pH in highly alkaline soils

Although plant growth is often limited at high pH, little is known about root-induced changes in the rhizospheres of plants growing in alkaline soils. The effect of Mn deficiency in Rhodes grass (Chloris gayana cv. Pioneer) and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. Rhizosphere pH was measured quantitatively, with a micro pH electrode, and qualitatively, with an agar/pH indicator solution. Manganese deficiency in Rhodes grass increased root-induced acidification of the rhizosphere in a soil profile in which N was supplied entirely as NO3-. Rhizosphere pH in the Mn deficient plants was up to 1.22 pH units lower than that of the bulk soil, while only 0.90 to 0.62 pH units lower in plants supplied with adequate Mn. When soil N was supplied entirely as NO3-, rhizosphere acidification was more efficient in inoculated lucerne (1.75 pH unit decrease) than in non-inoculated lucerne (1.16 pH unit decrease). This difference in capacity to lower rhizosphere pH is attributable to the ability of the inoculated lucerne to fix atmospheric N2 rather than relying on the soil N (NO3 ) reserves as the non-inoculated plants. Rhizosphere acidification in both Rhodes grass and lucerne was greatest in the meristematic root zone and least in the maturation root zone.

DNA packaging by L1 and L2 capsid proteins of bovine papillomavirus type 1

Encapsidation of circular DNA by papillomavirus capsid protein was investigated in Cos-1 cells. Plasmids carrying both an SV40 origin of replication (or) and an E. coli on were introduced into Cos-1 cells by DNA transfection. PV capsid proteins were supplied in trans by recombinant vaccinia viruses. Pseudovirions were purified from infected cells and their packaged DNA was extracted and used to transform E. coil as an indication of packaging efficacy. VLPs assembled from BPV-1 L1 alone packaged little plasmid DNA, whereas VLPs assembled from BPV-1 L1+L2 packaged plasmid DNA at least 50 times more effectively. BPV-1 L1+L2 VLPs packaged a plasmid containing BPV-1 sequence 8.2 +/- 3.1 times more effectively than a plasmid without BW sequences. Using a series of plasmid constructs comprising a core BPV-1 sequence and spacer DNA it was demonstrated that BW VLPs could accommodate a maximum of about 10.2 kb of plasmid DNA, and that longer closed circular DNA was truncated to produce less dense virions with shorter plasmid sequences. The present study suggests that packaging of genome within PV virions involves interaction of L2 protein with specific DNA sequences, and demonstrates that PV pseudovirions have the potential to be used as DNA delivery vectors for plasmids of up to 10.2 kb. (C) 1998 Academic Press.

Geochemical mass-balance and oxygen-isotope constrains on silcrete formation and its paleoclimatic implications in Southern Australia

Two geographically distinct silcrete associations are present in southern Australia, inland and eastern; these were sampled in central South Australia and central Victoria, respectively, At each site, both silicified and immediately adjacent unsilicified parent material were collected. Analytical data from these pairs were used to construct isocons, assuming Zr immobility, and to calculate the volume change and amount of silica introduced during silicification, These results, together with whole-rock oxygen isotope compositions, were used to determine the delta(18)O of th, introduced silica, The results show that the eastern silcretes in central Victoria are probably linked genetically to the associated basalts, weathering of which supplied the introduced silica, This conclusion is based on the close spatial connection between the two, as well as the substantial amount of introduced silica in the silcretes (greater than in the inland silcretes), resulting in volume increases in some eastern silcretes, Oxygen isotopic calculations for the silcretes indicate that the silica precipitated from groundwaters at temperatures slightly higher than present conditions. Silcrete formation apparently occurred during the Miocene and Pliocene (basalts in Victoria younger than Pliocene lack associated silcrete) and may reflect the much wetter climate in southeastern Australia at that time. The inland silcretes of central South Australia can be divided into pedogenic (the most common) and groundwater varieties. The pedogenic silcretes, which show typical soil features like columnar and nodular textures, contain moderate amounts of introduced silica that precipitated by evaporation from saline groundwaters, For the groundwater silcretes, which have massive textures and formed at or close to the water table, insufficient data are available to determine the mode of formation. The inland pedogenic silcretes have probably been farming from the Eocene-Miocene to the present, implying that conditions of seasonally high evaporation have occurred in central Australia during this time period. Thus silcrete formation depends on a complex interplay between climate and silica supply, and it is impossible to generalize that the presence of silcrete is indicative of a particular climate. Likewise, the elemental composition of silcretes, particularly Ti content, is not necessarily of climatic significance, Nevertheless, detailed geochemical and oxygen isotopic studies of a silcrete and its parent material can elucidate the mechanisms of silcrete formation, and if evaporation is indicated as a major factor in silcrete formation, then the climate at the time was likely to have been at least seasonally arid.

Description of Gluconacetobacter sacchari sp. nov., a new species of acetic acid bacterium isolated from the leaf sheath of sugar cane and from the pink sugar-cane mealy bug

A new species of the genus Gluconacetobacter, for which the name Gluconacetobacter sacchari sp. nov. is proposed, was isolated from the leaf sheath of sugar cane and from the pink sugar-cane mealy bug, Saccharicoccus sacchari, found on sugar cane growing in Queensland and northern New South Wales, Australia, The nearest phylogenetic relatives in the alpha-subclass of the Proteobacteria are Gluconacetobacter liquefaciens and Gluconacetobacter diazotrophicus, which have 98.8-99.3% and 97.9-98.5% 16S rDNA sequence similarity, respectively, to members of Gluconacetobacter sacchari. On the basis of the phylogenetic positioning of the strains, DNA reassociation studies, phenotypic tests and the presence of the Q10 ubiquinone, this new species was assigned to the genus Gluconacetobacter. No single phenotypic characteristic is unique to the species, but the species can be differentiated phenotypically from closely related members of the acetic acid bacteria by growth in the presence of 0.01% malachite green, growth on 30% glucose, an inability to fix nitrogen and an inability to grow with the L-amino acids asparagine, glycine, glutamine, threonine and tryptophan when D-mannitol was supplied as the sole carbon and energy source. The type strain of this species is strain SRI 1794(T) (= DSM 12717(T)).

Removal of a cysteine ligand from rubredoxin: assembly of Fe2S2 and Fe(S-Cys)(3)(OH) centres

The electron transfer protein rubredoxin from Clostridium pasteurianum contains an Fe(S-Cys)(4) active site. Mutant proteins C9G, C9A, C42G and C42A, in which cysteine ligands are replaced by non-ligating Gly or Ala residues, have been expressed in Escherichia coli. The C42A protein expresses with a (Fe2S2)-S-III cluster in place. In contrast, the other proteins are isolated in colourless forms, although a (Fe2S2)-S-III cluster may be assembled in the C42G protein via incubation with Fe-III and sulfide. The four mutant proteins were isolated as stable mononuclear Hg-II forms which were converted to unstable mononuclear Fe-III preparations that contain both holo and apo protein. The Fe-III systems were characterized by metal analysis and mass spectrometry and by electronic, electron paramagnetic resonance, X-ray absorption and resonance Raman spectroscopies. The dominant Fe-III form in the C9A preparation is a Fe(S-Cys)(3)(OH) centre, similar to that observed previously in the C6S mutant protein. Related centres are present in the proteins NifU and IscU responsible for assembly and repair of iron-sulfur clusters in both prokaryotic and eukaryotic cells. In addition to Fe(S-Cys)(3)(OH) centres, the C9G, C42G and C42A preparations contain a second four-coordinate Fe-III form in which a ligand appears to be supplied by the protein chain. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00775-0020355-1.

Neuroarchitecture of the color and polarization vision system of the stomatopod haptosquilla

The apposition compound eyes of stomatopod crustaceans contain a morphologically distinct eye region specialized for color and polarization vision, called the mid-band. In two stomatopod superfamilies, the mid-band is constructed from six rows of enlarged ommatidia containing multiple photoreceptor classes for spectral and polarization vision. The aim of this study was to begin to analyze the underlying neuroarchitecture, the design of which might reveal clues how the visual system interprets and communicates to deeper levels of the brain the multiple channels of information supplied by the retina. Reduced silver methods were used to investigate the axon pathways from different retinal regions to the lamina ganglionaris and from there to the medulla externa, the medulla interna, and the medulla terminalis. A swollen band of neuropil-here termed the accessory lobe-projects across the equator of. the lamina ganglionaris, the medulla externa, and the medulla interna and represents, structurally, the retina's mid-band. Serial semithin and ultrathin resin sections were used to reconstruct the projection of photoreceptor axons from the retina to the lamina ganglionaris. The eight axons originating from one ommatidium project to the same lamina cartridge. Seven short visual fibers end at two distinct levels in each lamina cartridge, thus geometrically separating the two channels of polarization and spectral information. The eighth visual fiber runs axially through the cartridge and terminates in the medulla externa. We conclude that spatial, color, and polarization information is divided into three parallel data streams from the retina to the central nervous system. (C) 2003 Wiley-Liss, Inc.

Fuel stagnation temperature effects on mixing with supersonic combustion flows

An experimental study of the effect of fuel stagnation temperature on mixing in a supersonic hydrogen-air flame is described, The combustor consisted of a constant-area rectangular duct with a centrally located fuel-injection strut that spanned the width. A high-enthalpy stream of air was supplied by a free-piston shock tunnel, and heated hydrogen fuel, supplied by a gun-tunnel, was injected into the freestream as a coflowing planar jet. The freestream total enthalpies were 5.6, 6.5, and 9 MJ/kg, and fuel stagnation temperatures were 300, 450, and 700 K, Raising the fuel stagnation temperature increased the fuel velocity to be near that of the airstream and resulted in a decrease in the mixing rate, Even as the fuel and air velocities became equal, significant mixing still occurred because of a large difference in density, Increasing the freestream enthalpy reduced the difference between the initial air temperature and the adiabatic flame temperature, which in turn reduced the heat addition, and subsequently, the amount of pressure rise in the duct.

The public's knowledge of and support for conservation of Australia's Tree-Kangaroos

After providing some brief background on Dendrolagus species in Australia, two consecutive surveys of Brisbane’s residents are used to assess public knowledge of tree-kangaroos and the stated degree of support for their conservation in Australia. The responses of participants in Survey I are based on their pre-survey knowledge of wildlife. The same additional set of participants completed Survey II after being provided with information on all the wildlife species mentioned in Survey I. Changes in the attitudes of respondents and their degree of support for the protection and conservation of Australia’s tree-kangaroos are measured, including changes in their contingent valuations and stated willingness to provide financial support for such conservation. Reasons for wanting to protect tree-kangaroos are specified and analyzed. Furthermore, changes that occur in the relative importance of these reasons with increased knowledge are also examined. Support for the conservation of tree-kangaroos is found to increase with the additional knowledge supplied. Furthermore, support for the conservation of Australia’s less well-known tropical mammals is shown to increase relative to better known mammals (icons) present in temperate areas, such as koalas and red kangaroos with this increased knowledge. Possible implications of the results for government conservation policies in Australia are examined.

Feasibility of an evidence-based literature search service for general practitioners

Objective: To test the feasibility of an evidence-based clinical literature search service to help answer general practitioners' (GPs') clinical questions. Design: Two search services supplied GPs who submitted questions with the best available empirical evidence to answer these questions. The GPs provided feedback on the value of the service, and concordance of answers from the two search services was assessed. Setting: Two literature search services (Queensland and Victoria), operating for nine months from February 1999. Main outcome measures: Use of the service; time taken to locate answers; availability of evidence; value of the service to GPs; and consistency of answers from the two services. Results: 58 GPs asked 160 questions (29 asked one, 11 asked five or more). The questions concerned treatment (65%), aetiology (17%), prognosis (13%), and diagnosis (5%). Answering a question took a mean of 3 hours 32 minutes of personnel time (95% Cl, 2.67-3.97); nine questions took longer than 10 hours each to answer, the longest taking 23 hours 30 minutes. Evidence of suitable quality to provide a sound answer was available for 126 (79%) questions. Feedback data for 84 (53%) questions, provided by 42 GPs, showed that they appreciated the service, and asking the questions changed clinical care. There were many minor differences between the answers from the two centres, and substantial differences in the evidence found for 4/14 questions. However, conclusions reached were largely similar, with no or only minor differences for all questions. Conclusions: It is feasible to provide a literature search service, but further assessment is needed to establish its cost effectiveness.

Experience with a low-cost telemedicine system in three developing countries

The Swinfen Charitable Trust was established in 1998 with the aim of helping the poor, sick and disabled in the developing world. It does this by setting up simple telemedicine links based on email to support doctors in isolated hospitals. The first telemedicine link was established to support the lone orthopaedic surgeon at the Centre for the Rehabilitation of the Paralysed (CRP) in Savar, near Dhaka in Bangladesh, in July 1999. An evaluation of the 27 referrals made during the first year of operation showed that the telemedical advice had been useful and cost-effective. Based on the success of the Bangladesh project, the Swinfen Charitable Trust supplied: digital cameras and tripods to more hospitals in other developing countries. These are Patan Hospital in Nepal (March 2000), Gizo Hospital in the Solomon Islands (March 2000), Helena Goldie Hospital: on New Georgia in the Solomon Islands (September 2000) and LAMB Hospital in Bangladesh (September 2000).

Heat of desorption measured by means of electrical heat excitation

A new method of measuring heat of desorption is proposed in this Letter. The principle of the method is to measure the amount of mass released when a controlled amount of energy is supplied directly to a solid adsorbent. This is in contrast to conventional methods such as microcalorimetry, where heat released upon adsorption is measured. In this method, a quantified heat supply is generated by passing a de current through a carbon pellet, which is equilibrated with a gas phase confined in a closed vessel. As a consequence of the heating, the particle temperature is increased, resulting in partial desorption of adsorbed molecules. The variations of pellet temperature and the system pressure with respect to time are used to determine the heat of desorption as a function of loading.

Controlled partial mine ventiliation recirculation trial at Mount Isa Mines

The ventilation and cooling of deep, hot mines present particular problems in Australia as a consequence of the surface climate, the size of the underground voids, the degree of mechanization and the cost of power in remote areas. A preliminary investigation of the effects of controlled partial recirculation of air was conducted in Mount Isa Mines' Deep Copper section. Gas and dust concentrations were measured in the exhaust air of the major working section to assess the potential for recirculating exhaust air to the intake airways to reduce the cost of providing an acceptable working environment in the deep parts of the mine. Studies were undertaken of airborne dust deposition in vertical airways and the efficiency of usage of the ventilation air in diluting contaminants. It was established that 45% of the respirable dust was deposited in a 130-m vertical raise and 60% of the air supplied to the section could be reused or recirculated. The first major field trial of a controlled partial recirculation system in Australia was undertaken in the light of these results and demonstrated excellent potential for significant reduction in ventilation costs. Gas and dust contaminant levels were well below the threshold limit values during the trial. It is concluded that controlled partial recirculation can be a practical, effective and safe aid to normal ventilation practice in Australian deep, hot mines.

The reduction of itch during burn wound healing

The purpose of this study was to find a method to reduce the itch experienced by patients who have sustained burn injuries, by using and comparing the effectiveness of 2 shower and bath oils. One product contained liquid paraffin with 5% colloidal oatmeal and the other contained liquid paraffin. The study was carried out in the Adult Burns Unit, Royal Brisbane Hospital (RBH), Brisbane. It was conducted during a 10-month period from July 1998 until April 1999. Thirty-five acute burns patients participated in an assessor-blind clinical trial. Patients were asked twice daily to rate their discomfort from itch and pain. The amount of antihistamine requested by each patient was totalled daily. Analysis of data supplied by patients showed that the group using the product with colloidal oatmeal reported significantly less itch and requested significantly less antihistamine than those using the oil containing liquid paraffin.