Cross-protective and Infection-Enhancing Immunity in Mice Vaccinated Against Flaviviruses Belonging to the Japanese Encephalitis Virus Serocomplex

The Japanese encephalitis virus serocomplex is a group of mosquito-borne flaviviruses that cause severe encephalitic disease in humans. The recent emergence of several members of this serocomplex in geographic regions where other closely related flaviviruses are endemic has raised urgent human health issues. Thus, the impact of vaccination against one of these neurotropic virus on the outcome of infection with a second, serologically related virus is unknown. We show here that immunity against Murray Valley encephalitis virus in vaccinated mice can cross-protect but also augment disease severity following challenge with Japanese encephalitis virus. Immunepotentiation of heterologous flavivirus disease was apparent in animals immunized with a 'killed' virus preparation when humoral antiviral immunty of low magnitude was elicited. (C) 2002 Elsevier Science Ltd. All rights reserved.

Investigation of candidate division TM7, a recently recognized major lineage of the domain bacteria with no known pure-culture representatives

A molecular approach was used to investigate a recently described candidate division of the domain Bacteria, TM7, currently known only from environmental 16S ribosomal DNA sequence data, A number of TM7-specific primers and probes were designed and evaluated. Fluorescence in situ hybridization (FISH) of a laboratory scale bioreactor using two independent TM7-specific probes revealed a conspicuous sheathed-filament morphotype, fortuitously enriched in the reactor. Morphologically, the filament matched the description of the Eikelboom morphotype 0041-0675 widely associated with bulking problems in activated-sludge wastewater treatment systems. Transmission electron microscopy of the bioreactor sludge demonstrated that the sheathed-filament morphotype had a typical gram-positive cell envelope ultrastructure. Therefore, TM7 is only the third bacterial lineage recognized to have gram-positive representatives. TM7-specific FISH analysis of two full-scale wastewater treatment plant sludges, including the one used to seed the laboratory scale reactor, indicated the presence of a number of morphotypes, including sheathed filaments. TM7-specific PCR clone libraries prepared from the two full-scale sludges yielded 23 novel TM7 sequences. Three subdivisions could be defined based on these data and publicly available sequences. Environmental sequence data and TM7-specific FISH analysis indicate that members of the TM7 division are present in a variety of terrestrial, aquatic, and clinical habitats. A highly atypical base substitution (Escherichia coli position 912; C to U) for bacterial 16S rRNAs was present in almost all TM7 sequences, suggesting that TM7 bacteria, like Archaea, may be streptomycin resistant at the ribosome level.

Caenorhabditis elegans mutants resistant to phosphine toxicity show increased longevity and cross-resistance to the synergistic action of oxygen

Phosphine (hydrogen phosphide, PH3) is the fumigant most widely used to protect stored products from pest infestation. Despite the importance of this chemical, little is known about its mode of action. We have created three phosphine-resistant lines (pre-1, pre-7, pre-33) in the model organism C. elegans, with LC50 values 2, 5, and 9 times greater than the fully susceptible parental strain. Molecular oxygen was shown to be an extremely effective synergist with phosphine as, under hyperoxic conditions, 100% mortality was observed in wild-type nematodes exposed to 0.1 mg/l phosphine, a nonlethal concentration in air. All three mutants were resistant to the synergistic effects of oxygen in proportion to their resistance to phosphine with one mutant, pre-33, showing complete resistance to this synergism. We take the proportionality of cross-resistance between phosphine and the synergistic effect of oxygen to imply that all three mutants circumvent a mechanism of phosphine toxicity that is directly coupled to oxygen metabolism. Compared with the wild-type strain, all three mutants have an extended average life expectancy of from 12.5 to 25.3%. This is consistent with the proposed involvement of oxidative stress in both phosphine toxicity and ageing. Because the wild-type and mutant nematodes develop at the same rate, the longevity is unlikely to be caused by a clk-type reduction in oxidative metabolism, a potential alternative mechanism of phosphine resistance.

Cross-border insolvency: Adoption of CLERP 8 as an evolution of Australian insolvency law

In 1997 the United Nations adopted the UNCITRAL Model Law on Cross-Border Insolvency and recommended that member states adopt it as part of domestic legislation. In 2002 Australia, an active participant in UNCITRAL's Working Group on Insolvency Law, announced that the next phase of the Commonwealth Government's Corporate Law Economic Reform Program would be a review of cross-border insolvency law. CLERP 8 seeks feedback on the proposed enactment of the Model Law by a separate Commonwealth statute. This article places such a development within the context of Australian cross-border insolvency law as it has evolved from early English bankruptcy legislation through case law arising from the banking collapses of the late 19th century to the more recent jurisprudence produced by corporate collapses of the late 1980s to early 1990s and current high-profile insolvencies.

Human melanoblasts in culture: Expression of BRN2 and synergistic regulation by fibroblast growth factor-2, stem cell factor, and endothelin-3

The BRN2 transcription factor (POU3F2, N-Oct-3) has been implicated in development of the melanocytic lineage and in melanoma. Using a low calcium medium supplemented with stem cell factor, fibroblast growth factor-2, endothelin-3 and cholera toxin, we have established and partially characterised human melanocyte precursor cells, which are unpigmented, contain immature melanosomes and lack L-dihydroxyphenylalanine reactivity. Melanoblast cultures expressed high levels of BRN2 compared to melanocytes, which decreased to a level similar to that of melanocytes when cultured in medium that contained phorbol ester but lacked endothelin-3, stem cell factor and fibroblast growth factor-2. This decrease in BRN2 accompanied a positive L-dihydroxyphenylalanine reaction and induction of melanosome maturation consistent with melanoblast differentiation seen during development. Culture of primary melanocytes in low calcium medium supplemented with stem cell factor, fibroblast growth factor-2 and endothelin-3 caused an increase in BRN2 protein levels with a concomitant change to a melanoblast-like morphology. Synergism between any two of these growth factors was required for BRN2 protein induction, whereas all three factors were required to alter melanocyte morphology and for maximal BRN2 protein expression. These finding implicate BRN2 as an early marker of melanoblasts that may contribute to the hierarchy of melanocytic gene control.

Effects of physical activity on emotional well-being among older Australian women - Cross-sectional and longitudinal analyses

Objective: To explore relationships between physical activity and mental health cross-sectionally and longitudinally in a large cohort of older Australian women. Method: Women in their 70s participating in the Australian Longitudinal Study on Women's Health responded in 1996 (aged 70-75) and in 1999 (aged 73-78). Cross-sectional data were analyzed for 10,063 women and longitudinal data for 6472. Self-reports were used to categorize women into four categories of physical activity at each time point as well as to define four physical activity transition categories across the 3-year period. Outcome variables for the cross-sectional analyses were the mental health component score (MCS) and mental health subscales of the Medical Outcomes Study Short Form (SF-36). The longitudinal analyses focused on changes in these variables. Confounders included the physical health component scale (PCS) of the SF-36, marital status, body mass index (BMI) and life events. Adjustment for baseline scores was included for the longitudinal analyses. Results: Cross-sectionally, higher levels of physical activity were associated with higher scores on all dependent variables, both with and without adjustment for confounders. Longitudinally, the effects were weaker, but women who had made a transition from some physical activity to none generally showed more negative changes in emotional well-being than those who had always been sedentary, while those who maintained or adopted physical activity had better outcomes. Conclusion: Physical activity is associated with emotional well-being among a population cohort of older women both cross-sectionally and longitudinally, supporting the need for the promotion of appropriate physical activity in this age group. (C) 2003 Elsevier Science Inc. All rights reserved.

Model-based analysis of anaerobic acetate uptake by a mixed culture of polyphosphate-accumulating and glycogen-accumulating organisms

An increasing number of studies shows that the glycogen-accumulating organisms (GAOs) can survive and may indeed proliferate under the alternating anaerobic/aerobic conditions found in EBPR systems, thus forming a strong competitor of the polyphosphate-accumulating organisms (PAOs). Understanding their behaviors in a mixed PAO and GAO culture under various operational conditions is essential for developing operating strategies that disadvantage the growth of this group of unwanted organisms. A model-based data analysis method is developed in this paper for the study of the anaerobic PAO and GAO activities in a mixed PAO and GAO culture. The method primarily makes use of the hydrogen ion production rate and the carbon dioxide transfer rate resulting from the acetate uptake processes by PAOs and GAOs, measured with a recently developed titration and off-gas analysis (TOGA) sensor. The method is demonstrated using the data from a laboratory-scale sequencing batch reactor (SBR) operated under alternating anaerobic and aerobic conditions. The data analysis using the proposed method strongly indicates a coexistence of PAOs and GAOs in the system, which was independently confirmed by fluorescent in situ hybridization (FISH) measurement. The model-based analysis also allowed the identification of the respective acetate uptake rates by PAOs and GAOs, along with a number of kinetic and stoichiometric parameters involved in the PAO and GAO models. The excellent fit between the model predictions and the experimental data not involved in parameter identification shows that the parameter values found are reliable and accurate. It also demonstrates that the current anaerobic PAO and GAO models are able to accurately characterize the PAO/GAO mixed culture obtained in this study. This is of major importance as no pure culture of either PAOs or GAOs has been reported to date, and hence the current PAO and GAO models were developed for the interpretation of experimental results of mixed cultures. The proposed method is readily applicable for detailed investigations of the competition between PAOs and GAOs in enriched cultures. However, the fermentation of organic substrates carried out by ordinary heterotrophs needs to be accounted for when the method is applied to the study of PAO and GAO competition in full-scale sludges. (C) 2003 Wiley Periodicals, Inc.

Growth of papaya nodal and rooted shoot cultures as affected by ACC, STS, culture vessel size and incubation conditions

Nodal shoot cultures of 'Clone 003', a selected Australian papaya cultivar, were cultured on modified De Fossard medium supplemented with chemicals that either promote ethylene evolution or inhibit action while in culture. Nodal shoot cultures grown in the presence of 1-aminocyclopropane carboxylic acid (ACC, 1.0 mM) resulted in a significant reduction in percent fresh and dry weights, shoot length, leaf area, petiole length and chlorophyll content, but leaf development was significantly increased. In contrast, nodal cultures grown in the presence of silver thiosulphate (STS, 0.5 mM) significantly produced the highest percentage of fresh and dry weights, shoot length, leaf production, leaf area expansion, petiole length and leaf chlorophyll content. Nodal cultures and rooted whole plantlets placed in medium-sized (125 mL) culture vessels had significantly better growth than those cultures placed in small (70 mL) or in large (250 mL) vessels. Cultures grown in medium-sized vessels had higher fresh and dry weights, longer shoots, more leaves and larger leaf area than those cultures placed in smaller or larger vessels. Similarly, values for said growth parameters and for chlorophyll content of the nodal and rooted whole plantlets were higher when they were incubated under high light intensity of 120 mumol m(-2)s(-1) at a prevailing temperature of either 20+/-1 C or 25+/-1 C.

Performing race, culture, and gender in an Indigenous Australian women's music and dance classroom

One perpetual concern among Indigenous Australian peoples is authenticity of voice. Who has the right to speak for, and to make representations about, the knowledges and cultures of Indigenous Australian peoples? Whose voice is more authentic, and what happens to these ways of knowing when they make the journey into mainstream Western academic classrooms? In this paper, I examine these questions within the politics of “doing” Indigenous Australian studies by focusing on my own experiences as a lecturer in the Aboriginal and Torres Strait Islander Studies Unit at the University of Queensland. My findings suggest that representation is a matter of problematizing positionality and, from a pedagogical standpoint, being aware of, and willing to address, the ways in which power, authority, and voice are performed and negotiated as teachers and learners of Indigenous Australian studies.

Sex- and race-related differences in cross-sectional geometry and bone density of the femoral mid-shaft in older adults

Background : Femoral shaft fracture incidence increases in older adults and is associated with low-energy trauma. Apart from bone density, the distribution and size of bone contributes to its strength. Aim : To examine if bone geometry and density of the femoral mid-shaft in older adults differs by sex and race, we studied 197 White women, 225 Black women, 242 White men, and 148 Black men aged 70-79 years participating in the Health, Aging, and Body Composition study; a prospective cohort study in the USA. A secondary purpose of the study was to examine the association of site-specific muscle and fat to bone geometry and density. Subjects and methods : Subjects were community-dwelling and reported no difficulty walking one-quarter of a mile or climbing stairs. Mid-femoral volumetric bone mineral density (vBMD, mg cm -3 ), total area (TA), cortical area (CA), medullary area (MA), cross-sectional moments of inertia (CSMI: I x , I y , J ), and muscle and fat areas (cm 2 ) were determined by computed tomography (CT; GE CT-9800, 10 mm slice thickness). Results : vBMD was greater in men than women with no difference by race ( p < 0.001). Bone areas and area moments of inertia were also greater in men than women ( p < 0.001), with Black women having higher values than White women for TA and CA. Standardizing geometric parameters for body size differences by dividing by powers of femur length did not negate the sex difference for TA and MA. Significant differences ( p < 0.05) among the four groups also remained for I x and J . Mid-thigh muscle area was an independent contributor to TA in all groups (Std beta = 0.181-0.351, p < 0.05) as well as CA in women (Std beta = 0.246-0.254, p < 0.01) and CSMI in White women (Std beta = 0.175-0.185, p < 0.05). Further, muscle area was a significant contributor to vBMD in Black women. Conclusion : These results indicate that bone geometry and density of the femoral diaphysis differs primarily by sex, rather than race, in older well-functioning adults. In addition, site-specific muscle area appears to have a potential contributory role to bone geometry parameters, especially in women.

Effect of time of harvest of budded virus on the selection of baculovirus FP mutants in cell culture

Rapid formation and selection of FP (few polyhedra) mutants occurs during serial passaging of Helicoverpa armigera nucleopolyhedrovirus (HaSNPV) in insect cell culture. The production of HaSNPV for use as biopesticides requires the passaging of the virus over a number of passages to produce enough virus inoculum for large-scale fermentation. During serial passaging in cell culture, FP mutants were rapidly selected, resulting in declined productivity and reduced potency of virus. Budded virus (BV) is usually harvested between 72 and 96 h postinfection (hpi) in order to obtain a high titer virus stock. In this study, the effect of tine of harvest (TOH) for BV on the selection rate of HaSNPV FP mutants during serial passaging was investigated. BV were harvested at different times postinfection, and each series was serially passaged for six passages. The productivity and percentage of FP mutants at each passage were determined. It was found that the selection of FP mutants can he reduced by employing an earlier TOH for BV. Serial passaging with BV harvested at 48 hpi showed a slower accumulation of FP mutants compared to that of BV harvested after 48 hpi. Higher cell specific yields were also maintained when BV were harvested at 48 hpi. When BV that were formed between 48 and 96 hpi were harvested and serially passaged, FP mutants quickly dominated the virus population. This suggests that the V formed and released between 48 and 96 hpi are most likely from FP mutant infected cells.

Cross-linking of PFA by electron beam irradiation

The effect of electron beam radiation on a perfluoroalkoxy (PFA) resin was examined using solid-state high-speed magic angle spinning F-19 NMR spectroscopy and FT-IR spectroscopy. Samples were prepared for analysis by subjecting them to electron beam radiation in the dose range 0.5-2.0 MGy at 633 K, which is above the crystalline melting temperature. The new structures were identified and include new saturated chain ends, short and long branches, unsaturated groups, and cross-links. The radiation chemical yield (G value) of new long branch points was greater than the G value of new chain ends, suggesting that cross-linking is the net radiolytic process. This conclusion was supported by an observed decrease in the crystallinity and an increase in the optical clarity of the polymer.