Visualising the activity of the cystine-glutamate antiporter in glial cells using antibodies to aminoadipic acid, a selectively transported substrate

The cystine-glutamate antiporter is a transport system that facilitates the uptake of cystine, concomitant with the release of glutamate. The cystine accumulated by this transporter is generally considered for use in the formation of the cysteine-containing antioxidant glutathione, which is abundant in many glial cells. This study used the simple strategy of generating an antibody to aminoadipic acid, a selective substrate for the cystine-glutamate antiporter. Stereospecific accumulation of aminoadipic acid into specific cell types in rat brain slice preparations was detected immunocytochemically. Strong accumulation was detected in astroglial cells in all brain regions studied including those in white matter tracts. Strong accumulation into radial glial cells, including the retinal Muller cells and the Bergmann glial cells was also observed. Glial accumulation was observed not only in cells within the blood brain barrier, but also outside such; anterior pituitary folliculostellate cell and intermediate lobe pituitary glial cells exhibited strong accumulation of aminoadipic acid. Interestingly, some glial cells such as the posterior pituitary glial cells (pituicytes) exhibited very little if any accumulation of aminoadipic acid. Within the brain labelling was not uniform. Particularly strong labelling was noted in some regions, such as the glial cells surrounding the CA1 pyramidal cells. By contrast, neurons never exhibited uptake of aminoadipic acid. Because cystine uptake is associated with glutamate release, it is suggested that this antiporter might contribute to release of glutamate from glial cells under some pathophysiological conditions. (C) 2001 Wiley-Liss, Inc.

Investigating technical and scale efficiencies of Australian universities through data envelopment analysis

Performance indicators in the public sector have often been criticised for being inadequate and not conducive to analysing efficiency. The main objective of this study is to use data envelopment analysis (DEA) to examine the relative efficiency of Australian universities. Three performance models are developed, namely, overall performance, performance on delivery of educational services, and performance on fee-paying enrolments. The findings based on 1995 data show that the university sector was performing well on technical and scale efficiency but there was room for improving performance on fee-paying enrolments. There were also small slacks in input utilisation. More universities were operating at decreasing returns to scale, indicating a potential to downsize. DEA helps in identifying the reference sets for inefficient institutions and objectively determines productivity improvements. As such, it can be a valuable benchmarking tool for educational administrators and assist in more efficient allocation of scarce resources. In the absence of market mechanisms to price educational outputs, which renders traditional production or cost functions inappropriate, universities are particularly obliged to seek alternative efficiency analysis methods such as DEA.

Vascular smooth muscle cell phenotypic modulation in culture is associated with reorganisation of contractile and cytoskeletal proteins

Smooth muscle cells (SMC) exhibit a functional plasticity, modulating from the mature phenotype in which the primary function is contraction, to a less differentiated state with increased capacities for motility, protein synthesis, and proliferation. The present study determined, using Western analysis, double-label immunofluorescence and confocal microscopy, whether changes in phenotypic expression of rabbit aortic SMC in culture could be correlated with alterations in expression and distribution of structural proteins. Contractile state SMC (days 1 and 3 of primary culture) showed distinct sorting of proteins into subcellular domains, consistent with the theory that the SMC structural machinery is compartmentalised within the cell. Proteins specialised for contraction (alpha -SM actin, SM-MHC, and calponin) were highly expressed in these cells and concentrated in the upper central region of the cell. Vimentin was confined to the body of the cell, providing support for the contractile apparatus but not co-localising with it. In line with its role in cell attachment and motility, beta -NM actin was localised to the cell periphery and basal cortex. The dense body protein alpha -actinin was concentrated at the cell periphery, possibly stabilising both contractile and motile apparatus. Vinculin-containing focal adhesions were well developed, indicating the cells' strong adhesion to substrate. In synthetic state SMC (passages 2-3 of culture), there was decreased expression of contractile and adhesion (vinculin) proteins with a concomitant increase in cytoskeletal proteins (beta -non-muscle [NM] actin and vimentin). These quantitative changes in structural proteins were associated with dramatic chan-es in their distribution. The distinct compartmentalisation of structural proteins observed in contractile state SMC was no longer obvious, with proteins more evenly distributed throughout die cytoplasm to accommodate altered cell function. Thus, SMC phenotypic modulation involves not only quantitative changes in contractile and cytoskeletal proteins, but also reorganisation of these proteins. Since the cytoskeleton acts as a spatial regulator of intracellular signalling, reorganisation of the cytoskeleton may lead to realignment of signalling molecules, which, in turn, may mediate the changes in function associated with SMC phenotypic modulation. (C) 2001 Wiley-Liss, Inc.

Work domain analysis and sensors II: Pasteurizer II case study

In this paper we use sensor-annotated abstraction hierarchies (Reising & Sanderson, 1996, 2002a,b) to show that unless appropriately instrumented, configural displays designed according to the principles of ecological interface design (EID) might be vulnerable to misinterpretation when sensors become unreliable or are unavailable. Building on foundations established in Reising and Sanderson (2002a) we use a pasteurization process control example to show how sensor-annotated AHs help the analyst determine the impact of different instrumentation engineering policies on a configural display that is part of an ecological interface. Our analyses suggest that configural displays showing higher-order properties of a system are especially vulnerable under some conservative instrumentation configurations. However, sensor-annotated AHs can be used to indicate where corrective instrumentation might be placed. We argue that if EID is to be effectively employed in the design of displays for complex systems, then the information needs of the human operator need to be considered while instrumentation requirements are being formulated. Rasmussen's abstraction hierarchy-and particularly its extension to the analysis of information captured by sensors and derived from sensors-may therefore be a useful adjunct to up-stream instrumentation design. (C) 2002 Elsevier Science Ltd. All rights reserved.

Work domain analysis and sensors I: principles and simple example

In this paper we establish a foundation for understanding the instrumentation needs of complex dynamic systems if ecological interface design (EID)-based interfaces are to be robust in the face of instrumentation failures. EID-based interfaces often include configural displays which reveal the higher-order properties of complex systems. However, concerns have been expressed that such displays might be misleading when instrumentation is unreliable or unavailable. Rasmussen's abstraction hierarchy (AH) formalism can be extended to include representations of sensors near the functions or properties about which they provide information, resulting in what we call a sensor-annotated abstraction hierarchy. Sensor-annotated AHs help the analyst determine the impact of different instrumentation engineering policies on higher-order system information by showing how the data provided from individual sensors propagates within and across levels of abstraction in the AH. The use of sensor-annotated AHs with a configural display is illustrated with a simple water reservoir example. We argue that if EID is to be effectively employed in the design of interfaces for complex systems, then the information needs of the human operator need to be considered at the earliest stages of system development while instrumentation requirements are being formulated. In this way, Rasmussen's AH promotes a formative approach to instrumentation engineering. (C) 2002 Elsevier Science Ltd. All rights reserved.

Opalescence in Australian-grown pecan kernels: Occurrence and causes

Opalescence is an unattractive browning of the interior of the pecan kernel compared to the white interior of normal kernels. The discoloration is due to the presence of free oil, resulting from decompartmentalization in the endosperm of opalescent,pecans. Using a subjective scoring system, approximately 70% of Australian-grown pecan kernels tested were found to exhibit opalescence to some degree. Evaluation of kernels for opalescence during the harvesting-processing chain showed that opalescence first becomes evident in kernels after mechanical cracking. Opalescent kernels were found to have lower levels of calcium and higher amounts of oil compared to nonoptalescent kernels. Differential scanning calorimetry showed that kernels do not freeze at -18 degreesC.

Friction factors and rheological behavior of Australian honey in a straight pipe

The pumping characteristics of four Australian honey samples were investigated in a straight pipe. Six flow rates (100-500 kg h(-1)) were studied at three temperatures (35-50degreesC). The pressure loss increased with an increase in the length of the pipe, as the low rate was increased and as the temperature was reduced. In the 25.4 mm-pipe, the Reynolds number ranged from 0.2-32.0 and are substantially less than the critica value (2040-2180) for laminar condition in the system. The relationship between the wall shear stress and shear rate approximated power-law behaviour, and the power-law index was not significantly (p>0.05) different from 1.0. The honey samples exhibited Newtonian behaviour at all the temperatures and this was confirmed by rheometric studies using Couette geometry. A friction chart was generated independent of temperature and the type of honey. An equation was developed to predict the pressure loss of the honey in a typical pipeline at any temperature once the viscosity-temperature relationship had been established.

Vasodilator mechanisms in the dorsal aorta of the giant shovelnose ray, Rhinobatus typus (Rajiformes; Rhinobatidae)

This study investigated the nature of vasodilator mechanisms in the dorsal aorta of the giant shovelnose ray, Rhinobatus typus. Anatomical techniques found no evidence for an endothelial nitric oxide synthase, but neural nitric oxide synthase was found to be present in the perivascular nerve fibres of the dorsal aorta and other arteries and veins using both NADPH-diaphorase staining and immunohistochemistry with a specific neural NOS antibody. Arteries and veins both contained large nNOS-positive nerve trunks from which smaller nNOS-positive bundles branched and formed a plexus in the vessel wall. Single, varicose nNOS-positive nerve fibres were present in both arteries and veins. Within the large bundles of both arteries and veins, groups of nNOS-positive cell bodies forming microganglia were observed. Double-labelling immunohistochemistry using an antibody to tyrosine hydroxylase showed that nearly all the NOS nerves were not sympathetic. Acetylcholine always caused constriction of isolated rings of the dorsal aorta and the nitric oxide donor, sodium nitroprusside, did not mediate any dilation. Addition of nicotine (3 x 10(-4) M) to preconstricted rings caused a vasodilation that was not affected by the nitric oxide synthase inhibitor, L-NNA (10(-4) M), nor the soluble guanylyl cyclase inhibitor, ODQ (10(-5) M). This nicotine-mediated vasodilation was, therefore, not due to the synthesis and release of NO. Disruption of the endothelium significantly reduced or eliminated the nicotine-mediated vasodilation. In addition. indomethacin (10(-5) M), an inhibitor of cyclooxygenases, significantly increased the time period to maximal dilation and reduced, but did not completely inhibit the nicotine-mediated vasodilation. These data support the hypothesis that a prostaglandin is released from the vascular endothelium of a batoid ray, as has been described previously in other groups of fishes. The function of the nitrergic innervation of the blood vessels is not known because nitric oxide does not appear to regulate vascular tone. (C) 2003 Elsevier Inc. All rights reserved.

Rapid quantification of hepatitis B virus DNA by real-time PCR using efficient TaqMan probe and extraction of virus DNA

Aim: To rapidly quantify hepatitis B virus (HBV) DNA by real-time PCR using efficient TaqMan probe and extraction methods of virus DNA. Methods: Three standards were prepared by cloning PCR products which targeted S, C and X region of HBV genome into pGEM-T vector respectively. A pair of primers and matched TaqMan probe were selected by comparing the copy number and the Ct values of HBV serum samples derived from the three different standard curves using certain serum DNA. Then the efficiency of six HBV DNA extraction methods including guanidinium isothiocyanate, proteinase K, NaI, NaOH lysis, alkaline lysis and simple boiling was analyzed in sample A, B and C by real-time PCR. Meanwhile, 8 clinical HBV serum samples were quantified. Results: The copy number of the same HBV serum sample originated from the standard curve of S, C and X regions was 5.7 × 104/ mL, 6.3 × 102/mL and 1.6 × 103/mL respectively. The relative Ct value was 26.6, 31.8 and 29.5 respectively. Therefore, primers and matched probe from S region were chosen for further optimization of six extraction methods. The copy number of HBV serum samples A, B and C was 3.49 × 109/mL, 2.08 × 106/mL and 4.40 × 107/mL respectively, the relative Ct value was 19.9, 30 and 26.2 in the method of NaOH lysis, which was the efficientest among six methods. Simple boiling showed a slightly lower efficiency than NaOH lysis. Guanidinium isothiocyanate, proteinase K and NaI displayed that the copy number of HBV serum sample A, B and C was around 105/ mL, meanwhile the Ct value was about 30. Alkaline failed to quantify the copy number of three HBV serum samples, Standard deviation (SD) and coefficient variation (CV) were very low in all 8 clinical HBV serum samples, showing that quantification of HBV DNA in triplicate was reliable and accurate. Conclusion: Real-time PCR based on optimized primers and TaqMan probe from S region in combination with NaOH lysis is a simple, rapid and accurate method for quantification of HBV serum DNA. © 2006 The WJG Press. All rights reserved.

Fluidization characteristics of moist food particles

Changes in fluidization behaviour of green peas particulates with change in moisture content during drying were investigated using a fluidized bed dryer. All drying experiments were conducted at 50 + 2 0C and 13 + 2 % RH using a heat pump dehumidifier system. Fluidization experiments were undertaken for the bedheights of 100, 80, 60 and 40 mm and at 10 moisture content levels. Fluidization behaviour was best fitted to the linear model of Umf = A + B m. A generalized model was also formulated using the height variation. Also generalized equation and Ergun equation was used to compare minimum fluidization velocity. Copyright ©2006 The Berkeley Electronic Press. All rights reserved.

Doom and gloom or a time for optimism: Potential aspirants' views about school leadership - now and for the future

Principal recruitment has attracted national and international attention in recent years (eg. Barty et al, 2005 in Australia; Earley et al, 2002 in the UK; Brooking et al, 2003 in New Zealand; Williams, 2003 in Canada). Importantly, Australian research in both state and non-state schools suggests that potential principal aspirants are less enthusiastic than might be expected in their desire to become principals (D’Arbon et al, 2002; Cranston et al, 2004; Lacey, 2002). Given the importance of ensuring we have quality leaders for our schools in the future, the research reported here (which is on-going) examined the views of potential aspirants (primary and secondary deputy principals) from one large government education system in Australia about the principalship and their intentions in seeking promotion (or otherwise) to such positions and the reasons driving these intentions. Data were collected via the Aspiring Principals Questionnaire (APQ) – especially developed for the study – comprising 38 closed items mainly of a Likert-type format, 5 open-ended items linked to particular closed items allowing participants to add their own suggestions/ideas, expand/elaborate on responses; and 4 further more general open-ended items. A number of system-level policy and practice recommendations have been developed from the findings.