Purification and membrane reconstitution of catalytically active Menkes copper-transporting P-type ATPase (MNK; ATP7A)

The MNK (Menkes disease protein; ATP7A) is a major copper- transporting P-type ATPase involved in the delivery of copper to cuproenzymes in the secretory pathway and the efflux of excess copper from extrahepatic tissues. Mutations in the MNK (ATP7A) gene result in Menkes disease, a fatal neurodegenerative copper deficiency disorder. Currently, detailed biochemical and biophysical analyses of MNK to better understand its mechanisms of copper transport are not possible due to the lack of purified MNK in an active form. To address this issue, we expressed human MNK with an N-terminal Glu-Glu tag in Sf9 [Spodoptera frugiperda (fall armyworm) 9] insect cells and purified it by antibody affinity chromatography followed by size-exclusion chromatography in the presence of the non-ionic detergent DDM (n-dodecyl b-D-maltopyranoside). Formation of the classical vanadate-sensitive phosphoenzyme by purified MNK was activated by Cu(I) [EC50=0.7 µM; h (Hill coefficient) was 4.6]. Furthermore, we report the first measurement of Cu(I)-dependent ATPase activity of MNK (K0.5=0.6 µM; h=5.0). The purified MNK demonstrated active ATP-dependent vectorial 64Cu transport when reconstituted into soya-bean asolectin liposomes. Together, these data demonstrated that Cu(I) interacts with MNK in a co-operative manner and with high affinity in the sub-micromolar range. The present study provides the first biochemical characterization of a purified full-length mammalian copper-transporting P-type ATPase associated with a human disease.

Novel data management algorithms in peer-to-peer content distribution networks

The peer-to-peer content distribution network (PCDN) is a hot topic recently, and it has a huge potential for massive data intensive applications on the Internet. One of the challenges in PCDN is routing for data sources and data deliveries. In this paper, we studied a type of network model which is formed by dynamic autonomy area, structured source servers and proxy servers. Based on this network model, we proposed a number of algorithms to address the routing and data delivery issues. According to the highly dynamics of the autonomy area, we established dynamic tree structure proliferation system routing, proxy routing and resource searching algorithms. The simulations results showed that the performance of the proposed network model and the algorithms are stable.

Estimating mean exposures from censored data: exposure to benzene in the Australian petroleum industry

A retrospective assessment of exposure to benzene was carried out for a nested case control study of lympho-haematopoietic cancers, including leukaemia, in the Australian petroleum industry. Each job or task in the industry was assigned a Base Estimate (BE) of exposure derived from task-based personal exposure assessments carried out by the company occupational hygienists. The BEs corresponded to the estimated arithmetic mean exposure to benzene for each job or task and were used in a deterministic algorithm to estimate the exposure of subjects in the study. Nearly all of the data sets underlying the BEs were found to contain some values below the limit of detection (LOD) of the sampling and analytical methods and some were very heavily censored; up to 95% of the data were below the LOD in some data sets. It was necessary, therefore, to use a method of calculating the arithmetic mean exposures that took into account the censored data. Three different methods were employed in an attempt to select the most appropriate method for the particular data in the study. A common method is to replace the missing (censored) values with half the detection limit. This method has been recommended for data sets where much of the data are below the limit of detection or where the data are highly skewed; with a geometric standard deviation of 3 or more. Another method, involving replacing the censored data with the limit of detection divided by the square root of 2, has been recommended when relatively few data are below the detection limit or where data are not highly skewed. A third method that was examined is Cohen's method. This involves mathematical extrapolation of the left-hand tail of the distribution, based on the distribution of the uncensored data, and calculation of the maximum likelihood estimate of the arithmetic mean. When these three methods were applied to the data in this study it was found that the first two simple methods give similar results in most cases. Cohen's method on the other hand, gave results that were generally, but not always, higher than simpler methods and in some cases gave extremely high and even implausible estimates of the mean. It appears that if the data deviate substantially from a simple log-normal distribution, particularly if high outliers are present, then Cohen's method produces erratic and unreliable estimates. After examining these results, and both the distributions and proportions of censored data, it was decided that the half limit of detection method was most suitable in this particular study.

Home-based resistance training is not sufficient to maintain improved glycemic control following supervised training in older individuals with type 2 diabetes

OBJECTIVE—To examine whether improvements in glycemic control and body composition resulting from 6 months of supervised high-intensity progressive resistance training could be maintained after an additional 6 months of home-based resistance training.

RESEARCH DESIGN AND METHODS—We performed a 12-month randomized controlled trial in 36 sedentary, overweight men and women with type 2 diabetes (aged 60–80 years) who were randomly assigned to moderate weight loss plus high-intensity progressive resistance training (RT&WL group) or moderate weight loss plus a control program (WL group). Supervised gymnasium-based training for 6 months was followed by an additional 6 months of home-based training. Glycemic control (HbA_1c), body composition, muscle strength, and metabolic syndrome abnormalities were assessed at 0, 3, 6, 9, and 12 months.

RESULTS—Compared with the WL group, HbA_1c decreased significantly more in the RT&WL group (–0.8%) during 6 months of supervised gymnasium-based training; however, this effect was not maintained after an additional 6 months of home-based training. In contrast, the greater increase in lean body mass (LBM) observed in the RT&WL group compared with the WL group (0.9 kg, P < 0.05) after the gymnasium-based training tended to be maintained after the home-based training (0.8 kg, P = 0.08). Similarly, the gymnasium-based increases in upper body and lower body muscle strength in the RT&WL group were maintained over the 12 months (P < 0.001). There were no between-group differences for changes in body weight, fat mass, fasting glucose, or insulin at 6 or 12 months.

CONCLUSIONS—In older adults with type 2 diabetes, home-based progressive resistance training was effective for maintaining the gymnasium-based improvements in muscle strength and LBM but not glycemic control. Reductions in adherence and exercise training volume and intensity seem to impede the effectiveness of home-based training for maintaining improved glycemic control.

Disease fingerprinting with cDNA microarrays reveals distinct gene expression profiles in lethal type 1 and type 2 cytokine-mediated inflammatory reactions

Development of polarized immune responses controls resistance and susceptibility to many microorganisms. However, studies of several infectious, allergic, and autoimmune diseases have shown that chronic type-1 and type-2 cytokine responses can also cause significant morbidity and mortality if left unchecked. We used mouse cDNA microarrays to molecularly phenotype the gene expression patterns that characterize two disparate but equally lethal forms of liver pathology that develop in Schistosoma mansoni infected mice polarized for type-1 and type-2 cytokine responses. Hierarchical clustering analysis identified at least three groups of genes associated with a polarized type-2 response and two linked with an extreme type-1 cytokine phenotype. Predictions about liver fibrosis,  apoptosis, and granulocyte recruitment and activation generated by the microarray studies were confirmed later by traditional biological assays. The data show that cDNA microarrays are useful not only for determining  coordinated gene expression profiles but are also highly effective for molecularly “fingerprinting” diseased tissues. Moreover, they illustrate the potential of genome-wide approaches for generating comprehensive views on the molecular and biochemical mechanisms regulating infectious  disease pathogenesis.

Effects of dietary lipid type on muscle fatty acid composition, carcass leanness, and meat toughness in lambs.

Isonitrogenous amounts of two protein sources differing in rumen degradation rate and in lipid composition were fed to sheep with or without a rapidly fermentable cereal grain. The effects on intake, carcass leanness, and muscle fatty acid (FA) composition were examined. Thirty-eight crossbred wether lambs (9 mo, 35 to 48 kg) were allocated by stratified randomization to six treatment groups: 1) basal diet of alfalfa hay:oat hay (20:80) ad libitum = basal; 2) basal + lupin (358 g DM/d) = lupin; 3) basal + fish meal (168 g DM/d) = fish meal; 4) basal + barley (358 g DM/d) = barley; 5) basal + barley + lupin (179 + 179 g DM/d) = barley/lupin; or 6) basal + barley + fish meal (179 + 84 g DM/d) = barley/ fish meal. Lambs were fed individually. Dietary treatments were imposed for 8 wk, and the supplements were offered at 2-d intervals. Daily feed intake and weekly BW of lambs were recorded. At the end of the feeding period lambs were slaughtered after an overnight fast. Hot carcass weight (HCW) and fat depth (GR; total fat and muscle tissue depth at 12th rib, 110 mm from midline) were recorded. At 24 h postmortem samples of longissimus thoracis (LT) and longissimus lumborum (LL) muscles were taken from chilled (4 deg C) carcasses for the assessment of FA composition and meat tenderness, respectively. Lambs fed lupin or fish meal with or without barley had heavier slaughter weights (P < 0.004) and HCW (P < 0.001) than lambs fed basal or barley when initial BW was included as a covariate. The lupin diet also resulted in heavier carcasses (P < 0.05) than the fish meal or barley/fish meal diets. With GR as an indicator, fish meal and barley/ fish meal diets produced leaner carcasses (P < 0.01) than lupin and barley/lupin lambs. Long-chain n-3 FA content [20:5n-3 (P < 0.001), 22:5n-3 (P < 0.003), and 22:6n-3 (P < 0.001)] in the LT muscle were substantially higher with the fish meal and barley/fish meal diets, whereas muscle total n-6 FA was increased (P < 0.003) by lupin and barley/lupin compared with all other diets. Thus, increased muscle long-chain n-3 FA content occurred without an increase in fatness measured as GR, whereas increased muscle n-6 FA content was associated with an increase in carcass fatness. Under these circumstances, a reduction in carcass fatness had no effect on meat tenderness measured as Warner-Bratzler shear force.

An agent-based framework for petroleum information services from distributed heterogeneous data resources

For making good decisions in the area of petroleum production, it is becoming a big problem how to timely gather sufficient and correct information, which may be stored in databases, data files, or on the World Wide Web. In this paper, Gaia methodology and Open Agent Architecture were employed to contribute a framework to solve above problem. The framework consists of three levels, namely, role mode, agent type, and agent instance. The model with five roles is analyzed. Four agent types are designed Six agent instances are developed for constructing the system of petroleum information services. The experimental results show that all agents in the system can work cooperatively to organize and retrieve relevant petroleum information. The successful implementation of the framework shows that agent-based technology can significantly facilitate the construction of complex systems in distributed heterogeneous data resource environment.

Bayesian classification of catchments using spatial data: a first step to improved modelling of catchment effects on stream ecological condition

A major challenge facing freshwater ecologists and managers is the development of models that link stream ecological condition to catchment scale effects, such as land use. Previous attempts to make such models have followed two general approaches. The bottom-up approach employs mechanistic models, which can quickly become too complex to be useful. The top-down approach employs empirical models derived from large data sets, and has often suffered from large amounts of unexplained variation in stream condition.

We believe that the lack of success of both modelling approaches may be at least partly explained by scientists considering too wide a breadth of catchment type. Thus, we believe that by stratifying large sets of catchments into groups of similar types prior to modelling, both types of models may be improved. This paper describes preliminary work using a Bayesian classification software package, ‘Autoclass’ (Cheeseman and Stutz 1996) to create classes of catchments within the Murray Darling Basin based on physiographic data.

Autoclass uses a model-based classification method that employs finite mixture modelling and trades off model fit versus complexity, leading to a parsimonious solution. The software provides information on the posterior probability that the classification is ‘correct’ and also probabilities for alternative classifications. The importance of each attribute in defining the individual classes is calculated and presented, assisting description of the classes. Each case is ‘assigned’ to a class based on membership probability, but the probability of membership of other classes is also provided. This feature deals very well with cases that do not fit neatly into a larger class. Lastly, Autoclass requires the user to specify the measurement error of continuous variables.

Catchments were derived from the Australian digital elevation model. Physiographic data werederived from national spatial data sets. There was very little information on measurement errors for the spatial data, and so a conservative error of 5% of data range was adopted for all continuous attributes. The incorporation of uncertainty into spatial data sets remains a research challenge.

The results of the classification were very encouraging. The software found nine classes of catchments in the Murray Darling Basin. The classes grouped together geographically, and followed altitude and latitude gradients, despite the fact that these variables were not included in the classification. Descriptions of the classes reveal very different physiographic environments, ranging from dry and flat catchments (i.e. lowlands), through to wet and hilly catchments (i.e. mountainous areas). Rainfall and slope were two important discriminators between classes. These two attributes, in particular, will affect the ways in which the stream interacts with the catchment, and can thus be expected to modify the effects of land use change on ecological condition. Thus, realistic models of the effects of land use change on streams would differ between the different types of catchments, and sound management practices will differ.

A small number of catchments were assigned to their primary class with relatively low probability. These catchments lie on the boundaries of groups of catchments, with the second most likely class being an adjacent group. The locations of these ‘uncertain’ catchments show that the Bayesian classification dealt well with cases that do not fit neatly into larger classes.

Although the results are intuitive, we cannot yet assess whether the classifications described in this paper would assist the modelling of catchment scale effects on stream ecological condition. It is most likely that catchment classification and modelling will be an iterative process, where the needs of the model are used to guide classification, and the results of classifications used to suggest further refinements to models.

Abstract art or the politics and pedagogies of getting read

The writing of academic abstracts is more than a tiresome necessity of scholarly life. It is a practice which goes beyond genre and technique to questions of identity and the promotional economies of academic work. In this paper we deconstruct a series of abstracts from a variety of refereed journals and conferences and develop a set of questions that allow us to 'read' the representation of data, argument, methodology and significance. We argue that the rules of abstract engagement are fluid and increasingly important with the advent of online journals and global citation indices. We suggest that abstract art is now an obligatory aspect of postgraduate supervision.

Interactive dance performance : movement data and the 'materiality' of the system

This paper describes a recent performance work I made using dance and live feed video
processing, 1 + x: mid-range projections, commissioned by the Seoul Contemporary Dance
Company and first performed in Melbourne in July 2005. This work forms a basis for discussing
my interest in creating performance images that reveal 'interiority'. I am interested in how you
embed the 'feel' of the human systematically in an interactive structure, and how that process
can produce a poetic that arises from the detailed and nuanced play between real and virtual
images on the same screen. How do you abstract and play with a performer's movement, play
with it in real and virtual time, so that it gives the work an emotional charge? Its like playing with
the process of 'becoming virtual' - and I'm being deliberately Deleuzian about that - how do you
'become virtual' in the sense of melding performer and image so that the meaning exists
between - in the connection between the two?

This quest to get the energy, the 'lived', 'felt' quality of the movement into the imagery gives rise
to research questions about how 'presence' is perceived in movement. What elements of the
raw movement data do you need to keep and what can you throwaway, and still keep the
personality, the emotion, the 'life' of that movement? How do you make a virtual, interactive
performance system that has its own 'materiality'?

Interpolatory type construction of general aggregation operators

In this chapter we examine a number of methods to construct aggregation operators of interpolatory type for specific applications. The construction is based on the desired values of the aggregation operator at certain prototypical points, and on other desired properties, such as, conjuctive, disjunctive or averaging behaviour, symmetry and marginals.

The mother-daughter guilt dynamic : effects of type 1 diabetes during life transitions

Aim. The aim of the study was to explore and describe the strategies young women with type 1 diabetes used to manage life transitions. The paper describes one aspect of how guilt dynamic often operates between mothers and daughters and how the women managed the guilt dynamic to create stability in their lives.
Background. When a child is diagnosed with diabetes, major transitional changes occur in the relationships between the mother and her child. The changes affect the psychological and social aspects of their lives and have a major impact on how young women manage their diabetes. A guilt dynamic between mothers and young women with diabetes emerged as a major theme in a larger study that investigated how young women with diabetes managed life transitions. Although the literature indicates that mothers of chronically ill children experience guilt feelings towards their children, little research was identified that addressed the emotional dynamics between mothers and daughters with diabetes.
Design. Using grounded theory method, interviews were conducted with 20 women with type 1 diabetes and five mothers during 2002 and 2003. Constant comparative analysis was used to analyse the data and develop an in-depth understanding of the experience of living with diabetes during life transitions.
Findings. The findings revealed that guilt feelings created a two-way dependency between mothers and their daughters with diabetes. The two-way dependency involved feelings of being a burden to each other, difficulty balancing responsibilities for diabetes management, difficulty relinquishing emotional and social dependency especially during life transitions. In addition, these issues were rarely discussed openly with each other or with health professionals. The findings provide additional information about the human experience of the mother–daughter relationship and the effect on coping with diabetes in the context of life transitions.
Conclusions. Understanding the impact diabetes has on the emotional and social well being of both women with type 1 diabetes and their mothers is critical in planning appropriate support for both groups. Most importantly, it is critical to understand the guilt dynamic that operates during young women with diabetes' life transitions when the daughters' dependency on their mother's control and responsibility for diabetes management undergo changes resulting in emotional responses, especially guilt feelings.
Relevance to clinical practice. Health professionals need to understand the emotional and social impact of the guilt dynamics between young women with type 1 diabetes and their mothers. Adequate and appropriate support can minimize the guilt feelings and enhance stability and quality of life for both mothers and their daughters, especially during major life transitions, such as motherhood.

Predictors of timely doctoral student completions by type of attendance: the utility of a pragmatic approach

Federal government changes to the funding of doctoral students have focused the attention of university management on their completion rates. The aims of this paper are to inform the allocation of institutional resources in a manner that improves the likelihood of timely doctoral completions and to highlight a process that can also be used for analyses of other key indicators of progression and attrition. The analyses and model development used national data readily available to all universities, which are collected in a standard approach through the Graduate Destinations Survey (GDS). The findings show that the most important variable for timely completion was attendance (full-time compared with part-time), whereby in terms of full-time equivalent (FTE) years of study, part-time students were far more likely to complete quickly than full-time students. For the full-time students, the key predictors of timely completion were residency, field of study and English-speaking background (ESB). The timeliness of part-time students was predicted by field of study and ESB. This study confirms that there is considerable variation by discipline for timely doctoral completions. The pragmatic application and prospective test of the derived models present a variety of opportunities for research student administrators. For example, those full-time students scoring highly represented a concentration of timely graduates more than 7.5 times higher than the lowest-scoring group - almost an order of magnitude of difference. In short, university management could gain tremendous value from more widely using the data available.

Predicting spatio-temporal distribution for eastern Australian birds using birds Australia's atlas data : survey method, habitat and seasonal effects

Faunal atlases are landscape-level survey collections that can be used for describing spatial and temporal patterns of distribution and densities. They can also serve as a basis for quantitative analysis of factors that may influence the distributions of species. We used a subset of Birds Australia’s Atlas of Australian Birds data (January 1998 to December 2002) to examine the spatio-temporal distribution patterns of 280 selected species in eastern Australia (17–37°S and 136–152°E). Using geographical information systems, this dataset was converted into point coverage and overlaid with a vegetation polygon layer and a half-degree grid. The exploratory data analysis involved calculating species-specific reporting rates spatially, per grid and per vegetation unit, and also temporally, by month and year. We found high spatio-temporal variability in the sampling effort. Using generalised linear models on unaggregated point data, the influences of four factors – survey method and month, geographical location and habitat type – were analysed for each species. When counts of point data were attributed to grid-cells, the total number of species correlated with the total number of surveys, while the number of records per species was highly variable. Surveys had high interannual location fidelity. The predictive values of each of the four factors were species-dependent. Location and habitat were correlated and highly predictive for species with restricted distribution and strong habitat preference. Month was only of importance for migratory species. The proportion of incidental sightings was important for extremely common or extremely rare species. In conclusion, behaviour of species differed sufficiently to require building a customized model for each species to predict distribution. Simple models were effective for habitat specialists with restricted ranges, but for generalists with wide distributions even complex models gave poor predictions.

Productivity growth and efficiency change in Malaysian manufacturing : recent evidence from disaggregated data

This paper estimates productivity growth in Malaysian manufacturing over the period 1983-1999. Malmquist productivity Indices (MPIs) have been computed using non parametric Data Envelopment Analysis (DEA) type linear programming, which show productivity growth sourced from efficiency change and growth in technology. Unlike previous studies, this study identifies the Malaysian manufacturing industries at the five digit breakdown of Malaysian Standard Industrial Classification (MSIC) thereby revealing more industry specific efficiency and technical growth patterns. Results indicate that two third of the industries (76 out of total 114 categories) experienced average annual
productivity improvement ranging from 0.1% to 7.8% over the sampled period. Average annual technical progress was recorded by 95 industry categories while technical efficiency improvement was achieved by 53 industries. Overall yearly average indicated relatively low productivity growth from the mid 1990’s onwards caused by either efficiency decline or technical regress. Summary results for industries reveal that some of the high rates of productivity growth have been recorded in glass and glass products (7.3%), Petroleum and coal (7.2%), industrial chemicals (4.9%) contributed from both efficiency improvement and technical progress ranging from 0.8% to 5.4% and from 1.7% to 4.1%, respectively. These results are expected to have some implications for ongoing and future strategic policy reform in Malaysian manufacturing generating a more sustainable growth for specific industry categories.