Combined in-fermenter extraction and cross-flow microfiltration for improved inclusion body processing

In this study we demonstrate a new in-fermenter chemical extraction procedure that degrades the cell wall of Escherichia coli and releases inclusion bodies (IBs) into the fermentation medium. We then prove that cross-flow microfiltration can be used to remove 91% of soluble contaminants from the released IBs. The extraction protocol, based on a combination of Triton X-100, EDTA, and intracellular T7 lysozyme, effectively released most of the intracellular soluble content without solubilising the IBs. Cross-flow microfiltration using a 0.2 mum ceramic membrane successfully recovered the granulocyte macrophagecolony stimulating factor (GM-CSF) IBs with removal of 91% of the soluble contaminants and virtually no loss of IBs to the permeate. The filtration efficiency, in terms of both flux and transmission, was significantly enhanced by infermenter Benzonase(R) digestion of nucleic acids following chemical extraction. Both the extraction and filtration methods exerted their efficacy directly on a crude fermentation broth, eliminating the need for cell recovery and re-suspension in buffer. The processes demonstrated here can all be performed using just a fermenter and a single cross-flow filtration unit, demonstrating a high level of process intensification. Furthermore, there is considerable scope to also use the microfiltration system to subsequently solubilise the IBs, to separate the denatured protein from cell debris, and to refold the protein using diafiltration. In this way refolded protein can potentially be obtained, in a relatively pure state, using only two unit operations. (C) 2004 Wiley Periodicals Inc.

Individual and societal bases of national identity - A comparative multi-level analysis

In a previous study (Jones and Smith, 1999) we established that much the same core pattern of national identity characterizes many developed countries. Using the national identity module from the 1995 International Social Survey Programme, we identified two dimensions of national identity: an ascriptive dimension resembling the concept of ethnic identity described in the historical and theoretical literature, and a voluntarist dimension closer to the notion of civic identity. Some writers view these dimensions in terms of a historical sequence but we find that both constructs coexist in the minds of individual respondents in the nations we examine (we exclude Bulgaria and the Philippines from the present but not the earlier analysis). The dataset used for the multilevel analyses reported here consists of 28 589 respondents in the remaining 21 countries included in the national identity database for the 1995 round of surveys. The macrosociological literature on national identity does not offer well-defined predictions about what precise patterns of national identification we might expect to find among the masses of the developed countries. There are, however, recurring themes from which one can construct plausible hypotheses about how countries might differ according to their level of development, broadly conceived. Thus, we hypothesize that forces such as post-industrialism and globalization tend to favour the more open voluntaristic form of national identity over the more restrictive ascribed form. We develop different multi-level models in order to evaluate specific hypotheses pertaining to such issues, by simultaneously relating individual and societal characteristics to the relative strength of individual commitment to these different types of national identity.

Reflections on onset-rime and phoneme sensitivity as predictors of beginning word reading

Rival claims have been made concerning the importance of rime sensitivity as a predictor of early word reading skill. Hulme et al. (2002) suggested that phoneme sensitivity is more strongly predictive of word reading ability than is onset-rime sensitivity. An examination of two independent data sets suggests that, although onset-rime sensitivity typically predicts school entrants' later word reading skill, phoneme sensitivity does predict more variation. However, multiple regression analyses do not reveal the level of phonological sensitivity that children need in order to understand alphabetic reading instruction. This issue is crucial to the detection of children at risk for reading failure and for the design of intervention programs for these children. A different analytic strategy is described for addressing this issue. (C) 2002 Elsevier Science (USA).

The essential work of fracture through the wall of uPVC pipe

The specific essential work of fracture, w(e), has been measured for a relatively thick walled uPVC pipe as a function of position through the wall of the pipe. w(e) was highest at the surface of the pipe and decreased significantly at the centre of the pipe wall. The variation in w(e) through the wall of the pipe correlated with the processing level of the uPVC material as measured by the critical temperature, T-c. The variability in the measured values of w(e) was substantially higher in the centre of the pipe where the processing levels were lower. This was likely to be a result of the variability in the microstructure of the material where poor processing had introduced regions of poor fusion of primary PVC particles. (C) 2002 Kluwer Academic Publishers.

Preparation and characterisation of biodegradable starch-based nanocomposite materials

Different formulations of biodegradable starch-polyester blend nanocomposite materials have been film blown on a pilot scale film blowing tower. The physical properties of different films have been examined by thermal and mechanical analysis and X-ray diffraction. The results show that the addition of an organoclay (from 0 to 5 wt%) significantly improves both the processing and tensile properties over the original starch blends. Wide angle X-ray diffraction (WAXD) results indicate that the best results were obtained for 30wt% starch blends, and the level of delamination depends on the ratio of starch to polyester and amount of organoclay added. The crystallisation temperature of the nanocomposite blends is significantly lower than the base blend. This is probably due to the platelets inhibiting order, and hence crystallisation, of the starch and polyester. The mechanical and thermal properties of the blends are also sensitive to the way the clay particles are dispersed. (C) 2003 Society of Chemical Industry.

Resistance to persuasive messages as a function of majority and minority source status

Three experiments examined the extent to which attitudes following majority and minority influence are resistant to counter-persuasion. In Experiment 1, participants' attitudes were measured after being exposed to two messages which argued opposite positions (initial pro-attitudinal message and subsequent, counter-attitudinal counter-message). Attitudes following minority endorsement of the initial message were more resistant to a (second) counter-message than attitudes following majority endorsement of the initial message. Experiment 2 replicated this finding when the message direction was reversed (counter-attitudinal initial message and pro-attitudinal counter-message) and showed that the level of message elaboration mediated the amount of attitude resistance. Experiment 3 included conditions where participants received only the counter-message and showed that minority-source participants had resisted the second message (counter-message) rather than being influenced by it. These results show that minority influence induces systematic processing of its arguments which leads to attitudes which are resistant to counter-persuasion. (C) 2003 Elsevier Science (USA). All rights reserved.

Endogenous Presentation of CD8+ T Cell Epitopes from Epstein-Barr Virus–encoded Nuclear Antigen 1

Epstein-Barr virus (EBV)-encoded nuclear antigen (EBNA)1 is thought to escape cytotoxic T lymphocyte (CTL) recognition through either self-inhibition of synthesis or by blockade of proteasomal degradation by the glycine-alanine repeat (GAr) domain. Here we show that EBNA1 has a remarkably varied cell type-dependent stability. However, these different degradation rates do not correspond to the level of major histocompatibility complex class I-restricted presentation of EBNA1 epitopes. In spite of the highly stable expression of EBNA1 in B cells, CTL epitopes derived from this protein are efficiently processed and presented to CD8(+) T cells. Furthermore, we show that EBV-infected B cells can readily activate EBNA1-specific memory T cell responses from healthy virus carriers. Functional assays revealed that processing of these EBNA1 epitopes is proteasome and transporter associated with antigen processing dependent. We also show that the endogenous presentation of these epitopes is dependent on the newly synthesized protein rather than the long-lived stable EBNA1. Based on these observations, we propose that defective ribosomal products, not the full-length antigen, are the primary source of endogenously processed CD8(+) T cell epitopes front EBNA1.

Financial and economic performance of long-rotation hardwood plantation investments in Queensland, Australia

In Queensland, Australia, there is presently a high level of interest in long-rotation hardwood plantation investments for sawlog production, despite the consensus in Australian literature that such investments are not financially viable. Continuing genetics, silviculture and processing research, and increasing awareness about the ecosystem services generated by plantations, are anticipated to make future plantings profitable and socio-economically desirable in many parts of Queensland. Financial and economic models of hardwood plantations in Queensland are developed to test this hypothesis. The economic model accounts for carbon sequestration, salinity amelioration and other ecosystem service values of hardwood plantations. A carbon model estimates the value of carbon sequestered, while salinity and other ecosystem service values are estimated by the benefit transfer method. Where high growth rates (20-25 m(3) ha(-1) year(-1)) are achievable, long-rotation hardwood plantations are profitable in Queensland Hardwood Regions 1, 3 and 7 when rural land values are less than $2300/ha. Under optimistic assumptions, hardwood plantations growing at a rate of 15 in 3 ha-1 year 1 are financially viable in Hardwood Regions 2, 4 and 8, provided land values are less than $1600/ha. The major implication of the economic analysis is that long-rotation hardwood plantation forestry is socio-economically justified in most Hardwood Regions, even though financial returns from timber production may be negative. (c) 2003 Elsevier B.V. All rights reserved.

Numerical simulation of magnetite segregation in a dense medium cyclone

Numerical simulations of turbulent driven flow in a dense medium cyclone with magnetite medium have been conducted using Fluent. The predicted air core shape and diameter were found to be close to the experimental results measured by gamma ray tomography. It is possible that the Large eddy simulation (LES) turbulence model with Mixture multi-phase model can be used to predict the air/slurry interface accurately although the LES may need a finer grid. Multi-phase simulations (air/water/medium) are showing appropriate medium segregation effects but are over-predicting the level of segregation compared to that measured by gamma-ray tomography in particular with over prediction of medium concentrations near the wall. Further, investigated the accurate prediction of axial segregation of magnetite using the LES turbulence model together with the multi-phase mixture model and viscosity corrections according to the feed particle loading factor. Addition of lift forces and viscosity correction improved the predictions especially near the wall. Predicted density profiles are very close to gamma ray tomography data showing a clear density drop near the wall. The effect of size distribution of the magnetite has been fully studied. It is interesting to note that the ultra-fine magnetite sizes (i.e. 2 and 7 mu m) are distributed uniformly throughout the cyclone. As the size of magnetite increases, more segregation of magnetite occurs close to the wall. The cut-density (d(50)) of the magnetite segregation is 32 gm, which is expected with superfine magnetite feed size distribution. At higher feed densities the agreement between the [Dungilson, 1999; Wood, J.C., 1990. A performance model for coal-washing dense medium cyclones, Ph.D. Thesis, JKMRC, University of Queensland] correlations and the CFD are reasonably good, but the overflow density is lower than the model predictions. It is believed that the excessive underflow volumetric flow rates are responsible for under prediction of the overflow density. (c) 2006 Elsevier Ltd. All rights reserved.

Effect of nutrition level and diets on creatinine excretion by sheep

In this experiment, creatinine (C) excretion by sheep was measured when they were fed different diets at different levels of intake. Creatinine excretion was not affected by the level of feed intake or the addition of salt to lucerne-based diets. However, differences between individual animals were significant. Creatinine excretion was significantly affected by diets, which were formulated by combining different amounts of lucerne chaff, oaten chaff and sorghum. It was also found that there were significant diurnal changes in the ratios of purine derivatives to creatinine (PD:C) in 3 hourly urine samples when the animals were fed either once or twice daily, but the average value for the PD:C ratio of any two urine samples taken 12 h apart was close to the daily mean. The results of this experiment suggest that if separate determination of the creatinine excretion by individual animals is made and the average value of the ratio of PD:C in two spot urine samples taken 12 h apart is used to predict PD excretion by spot urine sampling, microbial nitrogen flow can be estimated more accurately than when a fixed value of creatinine excretion is used for all animals and only a single urine sample is taken. (c) 2005 Elsevier B.V. All rights reserved.

Extrusion of mixtures of starch and D-limonene encapsulated with beta-cyclodextrin: Flavour retention and physical properties

d-Limonene was encapsulated with beta-cyclodextrin to improve its retention during pre-added flavour starch extrusion. The objective of this work was to determine the effect of processing condition on the flavour retention and extrudate properties. Corn starch containing five levels of beta-cyclodextrin-d-limonene capsules (0-5%) were extruded at five different maximum barrel temperatures (133-167 degrees C) and screw speeds (158-242 rpm) using a twin screw extruder. The effect of these parameters on the flavour retention, expansion, texture, colour difference (Delta E), Water Absorption Index, Water Solubility Index, and residence time distribution (RTD) were investigated. Barrel temperature and capsule level predominantly influenced flavour retention and extrudate properties, while screw speed primarily affected extruder performances such as torque, die pressure, specific mechanical energy and RTD. (c) 2005 Elsevier Ltd. All rights reserved.

Mechanical disruption of seagrass in the digestive tract of the dugong

The cheek teeth in dugongs are considered to be largely non-functional whereas the oral horny pads are important both in mechanical disruption of the diet and in conveying seagrass through the mouth. Particle size distributions of digesta from 41 dead stranded dugongs were examined to investigate the relationship between degree of food breakdown, gut region and functional surface area of the mouthparts. The in vitro ease of fracture of major dietary seagrass species were compared. The rate of food breakdown through the gut appears to be more closely linked to fibre level of the diet than to size or age of the dugong and its mouthparts. Low fibre seagrass, for example Halophila ovalis, breaks down at a faster rate than high fibre seagrass, for example Zostera capricorni both in dugong guts and in vitro. Several structural characteristics of seagrass, including level and arrangement of fibre, and water content, make it particularly amenable to mechanical breakdown. The soft mouthparts of the dugong are highly modified so that the entire oral cavity functions to crush low fibre seagrasses. Thus, the dugong has developed an efficient method of food ingestion and mastication that is suited to processing large quantities of soft seagrass during short dive times. The potential cost to the dugong in having lost its hard dental surfaces is that it has become restricted to a low fibre diet.

The effects of caffeine consumption on direct and indirect majority and minority influence

A study is reported that examines the effect of caffeine consumption on majority and minority influence. In a double blind procedure, 72 participants consumed an orange drink, which either contained caffeine (3.5mg per kilogram of body weight) or did not (placebo). After a 40-minute delay, participants read a counter-attitudinal message (antivoluntary euthanasia) endorsed by either a numerical majority or minority. Both direct (message issue, i.e., voluntary euthanasia) and indirect (message issue-related, i.e., abortion) change was assessed by attitude scales completed before and after exposure to the message. In the placebo condition, the findings replicated the predictions of Moscovici's (1980) conversion theory; namely, majorities leading to compliance (direct influence) and minorities leading to conversion (indirect influence). When participants had consumed caffeine, majorities not only led to more direct influence than in the placebo condition but also to indirect influence. Minorities, by contrast, had no impact on either level of influence. The results suggest that moderate levels of caffeine increase systematic processing of the message but the consequences of this vary for each source. When the source is a majority there was increased indirect influence while for a minority there was decreased indirect influence. The results show the need to understand how contextual factors can affect social influence processes.

Applying a Systems Approach to the Analysis of the Benefits of Conversion to Organic Production

Although the current level of organic production in industrialised countries amounts to little more than 1-2 percent, it is recognised that one of the major issues shaping agricultural output over the next several decades will be the demand for organic produce (Dixon et al. 2001). In Australia, the issues of healthy food and environmental concern contribute to increasing demand and market volumes for organic produce. However, in Indonesia, using more economical inputs for organic production is a supply-side factor driving organic production. For individual growers and processors, conversion from conventional to organic agriculture is often a challenging step, entailing a thorough revision of established practices and heightened market insecurity. This paper examines the potential for a systems approach to the analysis of the conversion process, to yield insights for household and community decisions. A framework for applying farming systems research to investigate the benefits of organic production in both Australia and Indonesia is discussed. The framework incorporates scope for farmer participation, crucial to the understanding of farming systems; analysis of production; and relationships to resources, technologies, markets, services, policies and institutions in their local cultural context. A systems approach offers the potential to internalise the external effects that may be constraining decisions to convert to organic production, and for the design of decision-making tools to assist households and the community. Systems models can guide policy design and serve as a mechanism for predicting the impact of changes to the policy and market environments. The increasing emphasis of farming systems research on community and environment in recent years is in keeping with the proposed application to organic production, processing and marketing issues. The approach will also facilitate the analysis of critical aspects of the Australian production, marketing and policy environment, and the investigation of these same features in an Indonesian context.

Changes in social identity when joining a new group: The role of individual-level and group-level adaptation processes

While classic intergroup theories have specified the processes explaining situational shifts in social identification, the processes whereby social identities change more profoundly and become integrated within the self have to be proposed. To this aim, the present studies investigate the processes by which group members integrate a new social identity as they are joining a new group. Combining a social identity approach and stress and coping models, this research tests if social factors (i.e., needs satisfied by fellow group members, social support), have an impact on the adaptation strategies group members use to deal with the novelty of the situation and to fit into their new group (seeking information & adopting group norms vs. disengaging). These strategies, in turn, should predict changes in level of identification with the new social group over time, as well as enhanced psychological adjustment. These associations are tested among university students over the course of their first academic year (Study 1), and among online gamers joining a newly established online community (Study 2). Path analyses provide support for the hypothesised associations. The results are discussed in light of recent theoretical developments pertaining to intraindividual changes in social identities and their integration in the self.