A rapid cell counting method utilising acridine orange as a novel discriminating marker for both cultured astrocytes and microglia

Cell culture analyses of growth, morphology and apoptosis commonly require counting of different cell types stained with antibodies to discriminate between them. Previously, we reported the use of l-Leucine methyl ester (l-LME) to prepare purified cultures of type 1 astrocytes with minimal microglia, and staining by GFAP and CD antibodies, respectively. Here, we demonstrate a novel use of acridine orange (AO) for rapid discrimination between these cell types using fluorescence microscopy. AO accumulates in the lysosomes and also binds strongly to nuclear DNA and cytoplasmic/nucleolar RNA. Microglia may contain abundant lysosomes due to known roles in homeostasis and immune response. AO staining of lysosomes was tested at a range of concentrations, and 2.5 μg/mL was most suitable. In agreement with previous reports, microglia treated with AO showed very intense yellow, orange or red granular cytoplasmic staining of lysosomes. Microglia contain a substantially higher number of lysosomes than astrocytes, which have a variable amount. We measured the microglia population at 5.14 ± 0.50% in mixed cultures. Thus, these results show AO is a novel discriminatory marker, as microglia were easily observed and counted in clumps on top of the monolayer of astrocytes, providing a rapid alternative to time-consuming and costly antibody-based assays.

Korea's developmental alliance : state, capital and the politics of rapid development

South Korea is often cited as a case of miraculous transformation from poverty to prosperity. Korea’s achievement of moving from one of the world’s poorest countries as recently as the early 1960s to the ranks of the ten biggest economies only four decades later has rightly attracted interest from policymakers and scholars alike.

This book identifies the factors that shaped relations between the state and big business in Korea, the ‘developmental alliance’. These factors offer a cogent framework in which to identify and predict changes in power relations between government and business. Rather than merely offering a means of explaining the rapid-growth phase of Korean development, the politics of the developmental alliance also help us understand how and why the Korean miracle turned to crisis in 1997 and why the subsequent recovery has been so uneven. In this way, the book highlights the political power of business, which is often underplayed in discussions of the development of Korea. It also sheds light on the constraints on policymakers during modernisation, and how power is shared among a small number of powerful parties.

Illustrating the tumultuous politics of the ‘developmental alliance’ between business and government during the rise and decline of South Korea’s economic miracle, this book is an essential read for anyone interested in Korean politcs, economics and development,

Plasma amino acid response after ingestion of different whey protein fractions

Background and objectives The digestion rate of proteins and subsequent absorption of amino acids can independently modulate protein metabolism. The objective of the present study was to examine the blood amino acid response to whey protein isolate (WPI), β-lactoglobulin-enriched WPI, hydrolysed WPI and a flavour-identical control.

Methods Eight healthy adults (four female, four male) were recruited (mean±standard error of the mean: age, 27.0±0.76 years; body mass index, 23.2±0.8 kg/cm2) and after an overnight fast consumed 500 ml of each drink, each containing 25g protein, in a cross-over design. Blood was taken at rest and then every 15 min for 2 h post ingestion.

Results Ingesting the β-lactoglobulin-enriched WPI drink resulted in significantly greater plasma leucine concentrations at 45-120 min and significantly greater branched-chain amino acid concentrations at 60-105 min post ingestion compared with hydrolysed WPI. No differences were observed between WPI and β-lactoglobulin-enriched WPI, and all protein drinks resulted in elevated blood amino acids compared with flavour-identical control.

Conclusions In conclusion, whole proteins resulted in a more rapid absorption of leucine and branched-chain amino acid into the blood compared with the hydrolysed molecular form of whey protein.

The changing face of literacy policy in Victoria

In the transition from the twentieth to the twenty-first century, literacy has undergone a fundamental change in the shift from page to screen as the dominant basis for communication. In a communications environment characterised by multimodality - integration of modes of linguistic, visual, audio, gestural and spatial modes of meaning - young people require a broadened repertoire of literacy capacities.
Educational authorities with responsibility for literacy policy have responded in terms of curriculum, and assessment advice within a context of rapidly changing forms of multimodal communication. This paper details the early twenty-first century response of one educational authoríty, the Department of Education, Victoria, in reviewing early years literacy curriculum and assessment in light of the rapid developments in digital communications.

Changes in skin colour and cortisol response of Australian snapper Pagrus auratus (Bloch & Schneider, 1801) to different background colours

A two-factor experiment was carried out to investigate the change in skin colour and plasma cortisol response of cultured Australian snapper Pagrus auratus to a change in background colour. Snapper (mean weight=437 g) were held in black or white tanks and fed diets containing 39 mg unesterified astaxanthin kg⁻¹ for 49 days before being transferred from white tanks to black cages (WB) or black tanks to white cages (BW). Skin colour values [L^* (lightness), a^* (redness) and b^* (yellowness)] of all snapper were measured at stocking (t=0 days) and from cages of fish randomly assigned to each sampling time at 0.25, 0.5, 1, 2, 3, 5 and 7 days. Plasma cortisol was measured in anaesthetized snapper following colour measurements at 0, 1 and 7 days. Fish from additional black-to-black (BB) and white-to-white (WW) control treatments were also sampled for colour and cortisol at those times. Rapid changes occurred in skin lightness (L^* values) after altering background colour with maximum change in L^* values for BW and WB treatments occurring within 1 day. Skin redness (a*) of BW snapper continued to steadily decrease over the 7 days (a*=7.93 × e^{−0.051 × time}). Plasma cortisol concentrations were highest at stocking when fish were held at greater densities and were not affected by cage colour. The results of this study suggest that transferring dark coloured snapper to white cages for 1 day is sufficient to affect the greatest benefit in terms of producing light coloured fish while minimizing the reduction in favourable red skin colouration.

Beyond mimesis : place, interpretation and response

My study examines the subjective nature of artistic interpretation through the notion of mimesis as process or transformation of material. Influential factors that mediate in the artistic process, such as memory, reflection and an awareness of cultural analogy and metaphor, are examined and related to a specific project in the studio, where the mediation process is further influenced by the materials used to produce the images. My studies of the concept of mimesis have revealed an intermediary realm that exists in the space between empirical reality and its interpretation. Throughout history the process of mimesis has been integral to all forms of the arts. In Plato's time the production of an image that simulated things as they appeared to the eye was considered a desired ideal. Aristotle later introduced developments which extended this concept to include a refiguring or reforming of material derived from the original source, making new connections between existing factors and in this transformation bringing new meanings to a symbolically constituted world. This discussion of the representation of reality, the influence of a dialogue between notions of imitation and the recreation of material continues throughout the exegesis. My study emphasises the interpretive stage of the mimetic process where a consideration of these themes is most relevant and some of the factors that can influence its outcome. It is my opinion that the production of images in response to the particularities of place can be defined in three stages. Firstly, the experience of the place; secondly, the beginning and maturation of the idea or concept; where mimesis takes place, and thirdly, the production of the art work in response. This process is illustrated in Part 2 of the exegesis, where the development of the studio work is documented and linked with the themes discussed in Part 1. The geographic site or place I selected to study is adjacent to Mt. Noorat, a volcanic site in the Western district of Victoria; the surrounding plains are littered with scoria that has been thrown out of the volcano thousands of years ago. Early British, Scottish and Irish settlers to this region used the stone to construct fences reminiscent of their homeland, through this activity they cleared the land and confined and protected their stock. My interests are in factors that include - the material of the stone, notions of enclosure and safety, of boundaries and circumscribed space, and of the cultural reflection that has taken place in this reconstruction of Eurocentric vision. These walls also represent the means by which land was enclosed and property defined, moving from a situation of public access to notions of ownership and the annexation of land for individual gain. Around each point of eruption, the craggy volcanic scoria has been used to create a constructed landscape which both symbolises and mirrors the Anglo - Celtic origins of the people. I have used the legend of Narcissus to illustrate the self-reflective and introspective processes that the settlers invoked in their attempts to come to terms with a strange land. I consider that the story of Narcissus, who fell in love with his own reflection, finds a parellel in the creation of the walls. The re-creation of artifacts from their own cultural environment provided the settlers with a familiar 'face' in an alien world; a reassurance of the familiar in an unfamiliar terrain. Part of this study is an investigation of this notion of landscape as cultural reflection. Geographers have long known that landscape is a cultural construct, an historically evolving ideal manifested in painting, prints and drawings as well as poetry, gardens and parks. One can view these constructions as illustrations or images of meaning which constitute representations of cultural ideals. The neo-classical influence reflected in the paintings of artists who accompanied the early expeditions to Australia demonstrates these themes. The medium of the mirror provides the opportunity to suggest aspects of a cultural reflection and an awareness of identity that has relevance to contemporary Australian culture, therefore, I have allowed it to play a major role throughout this study. Its role in mimesis, firstly, as a reflection in an imitative sense is established, then in its refigurative role, in which the similarities between the original and the reformed rely more on correlative factors than representation. I have used examples from the history of art to illustrate this potential. The formation and development of a narrative involving reflection threads throughout the thesis, both in the visual presentation and in the exegesis. The production of a body of paintings, drawings and sculpture reflect my interpretation and response to the particular site. The correspondences between these works and my theoretical concerns is articulated in the exegesis. The metaphor implied by the use of the walls as agents of enclosure also refers to the capacity of the individual to be confined by notional boundaries and restrictive practices where totalising systems of thought dominate theoretical debate and restrict its freedom. I have used images where gaps in the walls represent the potential implicit to the concept of liminal space, where the spectator moves from one physical space to another and from one stage of development to another. The threshold of this opening in the walls becomes the site where transformation can take place, a metaphor for the mimetic process where the initial experience is translated and transformed into the final product. The paintings, drawings and other works in this series fulfil the role of marks on the surface of the mirror, separating the initial experience from the processes of memory, reflection and speculation. The works draw attention to the materiality that they represent and yet provide the opportunity for new insights and experiences, allowing the subjective nature of artistic activity to combine symbolic elements relating to the site, resulting in the production of meaning.

The influence of specific luminal factors on the colonic epithelium : high-dose butyrate and physical changes suppress early carcinogenic events in rats

INTRODUCTION Although luminal delivery of butyrate is one putative mechanism by which biology of the colonic epithelium might be influenced by changes in luminal contents, there is a paucity of supportive cause–effect evidence. This study aimed to directly establish whether distal colonic butyrate delivery is able to alter the response of the distal colonic epithelium to a carcinogen.
METHODS Groups of male Sprague-Dawley rats with chronically intubated colons received infusions of 80 mM butyrate or 0.9 percent saline into distal colon two or five times daily. Three weeks after exposure to azoxymethane (15 mg/kg subcutaneously), the density of aberrant crypts was quantified in distal colon.
RESULTS Infusions of 0.5 ml twice daily, whether containing saline or butyrate, decreased the number of aberrant crypt foci by 45 percent compared with rats receiving no infusions (P = 0.004, analysis of variance). Similar results were obtained when infusions were restricted to the postinitiation phase. When infusions were increased to 1 ml five times daily, saline infusions similarly suppressed aberrant crypt formation (38 percent), but butyrate infusions suppressed it to a greater degree (by 64 percent; P = 0.02 compared with saline infusion, t-test).
CONCLUSIONS High levels of butyrate delivery to the distal colonic lumen alter the epithelial response to a carcinogen in otherwise healthy rats. This finding directly supports the notion that the effects of butyrate on cells in vitro do occur in vivo provided a sufficient dose is delivered. The effect of infusion of liquid per se on the epithelial response highlights the potential impact physical changes alone can have on the colonic epithelium.

Prevention of a chronic progressive form of experimental autoimmune encephalomyelitis by an antibody against mucosal addressin cell adhesion molecule-1, given early in the course of disease progression.

A role for α4 and β7 integrins in mediating leucocyte entry into the central nervous system in the multiple sclerosis (MS)-like disease experimental autoimmune encephalomyelitis (EAE) has been demonstrated. However, the individual contributions of their respective ligands mucosal addressin cell adhesion molecule-1 (MAdCAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-cadherin expressed on the blood-brain barrier has not been determined. In the present paper, it is shown that an antibody directed against MAdCAM-1, the preferential ligand for α4β7, effectively prevented the development of a progressive, non-remitting, form of EAE, actively induced by injection of myelin oligodendrocyte glycoprotein peptide (MOG(_35-55)) autoantigen. Combinational treatment with both anti-MAdCAM-1, VCAM-1, and intercellular adhesion molecule-1 (ICAM-1) (ligand for integrin lymphocyte function-associated antigen (LFA)-1) mAbs led to more rapid remission than that obtained with anti-MAdCAM-1 antibody alone. However, neither MAdCAM-1 monotherapy, nor combinational antibody blockade was preventative when administered late in the course of disease progression. In conclusion, MAdCAM-1 plays a major contributory role in the progression of chronic EAE and is a potential therapeutic target for the treatment of MS. Critically, antivascular addressin therapy must be given eaA role for alpha4 and beta7 integrins in mediating leucocyte entry into the central nervous system in the multiple sclerosis (MS)-like disease experimental autoimmune encephalomyelitis (EAE) has been demonstrated. However, the individual contributions of their respective ligands mucosal addressin cell adhesion molecule-1 (MAdCAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-cadherin expressed on the blood-brain barrier has not been determined. In the present paper, it is shown that an antibody directed against MAdCAM-1, the preferential ligand for alpha4beta7, effectively prevented the development of a progressive, non-remitting, form of EAE, actively induced by injection of myelin oligodendrocyte glycoprotein peptide (MOG(35-55)) autoantigen. Combinational treatment with both anti-MAdCAM-1, VCAM-1, and intercellular adhesion molecule-1 (ICAM-1) (ligand for integrin lymphocyte function-associated antigen (LFA)-1) mAbs led to more rapid remission than that obtained with anti-MAdCAM-1 antibody alone. However, neither MAdCAM-1 monotherapy, nor combinational antibody blockade was preventative when administered late in the course of disease progression. In conclusion, MAdCAM-1 plays a major contributory role in the progression of chronic EAE and is a potential therapeutic target for the treatment of MS. Critically, antivascular addressin therapy must be given early in the course of disease prior to the establishment of irreversible damage if it is to be effective, as a single treatment modality.

Evaluation of an early intervention program for childhood conduct problems

Treatment outcomes of an early intervention program for childhood conduct problems were evaluated. Results provided support for the longitudinal effectiveness of the program in reducing conduct problems, social difficulties, and psychosocial impairment for young primary school aged children. Children's experience and expression of anger also reduced across the program's duration. The portfolio presents four case studies whose results provide further support for Exposure and Response Prevention (ERP) as an effective and clinically useful treatment for paediatric OCD in everyday clinical contexts.

Well-being achievement in Pacific Asia

The world has undergone rapid and tremendous change in recent decades. While many nations have achieved ever-higher per capita incomes, and higher well-being according to traditional measures, they have also experienced profound internal change. This change has lead to widespread concerns regarding social exclusion, human security, levels of personal satisfaction and happiness. Other countries have faired much less well, as according to many well-being measures they are worse off than they were 10 or 20 years ago. Life expectancies, for example, have fallen dramatically in many countries and are likely to fall substantially in others. The incidence of income poverty is higher today in many countries than it was ten years ago. Worldwide, more than a billion people currently live on less than one dollar per day. Social science research on living standards, human well-being and quality of life has come a long way over recent years, altering in response to changing global conditions, new research priorities, new conceptualisations and improved data resources. Twenty five years ago, national well-being achievement comparisons relied very heavily, and in some circles exclusively, on measures of income per capita. The same exercise would today be based a range of indicators, including summary measures of human well-being such as the well-known Human Development Index (UNDP, 2005). This is consistent with the commonly accepted view that human well-being is best treated as a multidimensional concept along the lines advocated by Sen (1985, 1993), Stewart (1985), Doyal & Gough (1991), Ramsay (1992), Cummins (1996), Narayan et al. (2000) or Nussbaum (2000) and others, as summarised in Alkire (2002). This view tends not to reject the relevance of income based or economic measures per se, simply positing that there is more to well-being achievement than simply increasing incomes. The widespread acceptance that well-being is multidimensional has more recently been accompanied by another important recognition. This relates not so much to current levels of well-being, but to the likelihood of declines in future levels. This recognition has spawned a rapidly growing literature on what is now termed as ‘vulnerability’. The vulnerability literature has primarily been concerned with the likelihood of individuals falling below the poverty line, be it defined in terms of income, consumption or health. Among the influential early vulnerability studies are Ravallion (1998), Jalan & Ravallion (1998) and Dercon & Krishnan (1999), each of which distinguished between transient and chronic poverty.

Transient response analysis of steel in concrete

A new approach is presented for the analysis of galvanostatically induced transients allowing for the rapid evaluation of the corrosion activity of steel in concrete. This method of analysis is based on the iterative fitting of a non-exponential model based on a modified KWW (Kohlrausch–Williams–Watt) formalism. This analysis yields values for the parameters related to corrosion such as the concrete resistance, polarisation resistance, interfacial capacitance and β, the non-ideality exponent.

On the electrochemical response and interfacial properties of steel–Ca(OH)2 and the steel–concrete system measured using galvanostatic pulses

The interfacial properties of the steel–concrete system are examined via a new approach for evaluation of galvanostatic pulse data. This methodology allows for rapid determination of the corrosion activity of steel, and readily yields values for parameters related to corrosion such as the polarisation resistance and interfacial capacitance. The method of analysis is based on the iterative fitting of a non-exponential model based on a modified Kohlrausch–Williams–Watt (KWW) formalism. The transient behaviour of steel in concrete is non-exponential in its form and, when analysed this way, an exponent β can be determined characterising the exponential non-ideality of the transient. This non-ideality parameter is found to differ significantly for actively corroding and passive specimens, thereby serving as a useful index to the level of corrosion being experienced. Furthermore, the investigation of the interfacial characteristics of the system, previously unobtainable in a reproducible manner via other electrochemical methods, reveal information regarding the kinetic factors governing corrosion of steel in concrete.

Evidence and implications for early intervention in bipolar disorder

Aims: To review the evidence that supports early intervention in the treatment of bipolar disorder.

Background: Bipolar disorder is a pleomorphic condition, with varying manifestations that are determined by a number of complex factors including the ‘‘stage’’ of illness. It is consequently a notoriously difficult illness to diagnose and as a corollary is associated with lengthy delays in recognition and the initiation of suitable treatment.

Methods: A literature search was conducted using MEDLINE augmented by a manual search.

Results: Emerging neuroimaging data suggests that, in contrast to schizophrenia, where at the time of a first-episode of illness there is already discernible volume loss, in bipolar disorder, gross brain structure is relatively preserved, and it is only with recurrences that there is a sequential, but marked loss of brain volume. Recent evidence suggests that both pharmacotherapy and psychotherapy are more effective if instituted early in the course of bipolar disorder, and that with multiple episodes and disease progression there is a noticeable decline in treatment response.

Conclusions: Such data supports the notion of clinical staging, and the tailored implementation of treatments according to the stage of illness. The progressive nature of bipolar disorder further supports the concept that the first episode is a period that requires energetic broad-based treatment, with the hope that this could alter the temporal trajectory of the illness. It also raises hope that prompt treatment may be neuroprotective and that this perhaps attenuates or even prevents the neurostructural and neurocognitive changes seen to emerge with chronicity. This highlights the need for early identification at a population level and the necessity of implementing treatments and services at a stage of the illness where prognosis is optimal.

The disappearing mammal fauna of northern Australia : context, cause, and response

This article provides a context to, attempts an explanation for, and proposes a response to the recent demonstration of rapid and severe decline of the native mammal fauna of Kakadu National Park. This decline is consistent with, but might be more accentuated than, declines reported elsewhere in northern Australia; however, such a comparison is constrained by the sparse information base across this region. Disconcertingly, the decline has similarities with the earlier phase of mammal extinctions that occurred elsewhere in Australia. We considered four proximate factors (individually or interactively) that might be driving the observed decline: habitat change, predation (by feral cats), poisoning (by invading cane toads), and novel disease. No single factor readily explains the current decline. The current rapid decline of mammals in Kakadu National Park and northern Australia suggests that the fate of biodiversity globally might be even bleaker than evident in recent reviews, and that the establishment of conservation reserves alone is insufficient to maintain biodiversity. This latter conclusion is not new; but the results reported here further stress the need to manage reserves far more intensively, purposefully, and effectively, and to audit regularly their biodiversity conservation performance.

Identification and genetic determination of an early life risk disposition for depressive disorder : atypical stress-related behaviour in early childhood

Progress in psychiatric genetics has been slow despite evidence of high heritability for most mental disorders. We argue that greater use of early detectable intermediate traits (endophenotypes) with the highest likely aetiological significance to depression, rather than complex clinical phenotypes, would be advantageous. Longitudinal data from the Western Australian Pregnancy Cohort (Raine) Study were used to identify an early life behavioural endophenotype for atypical hypothalamic-pituitaryadrenocortical function in adolescence, a neurobiological indicator of anxiety and depression. A set of descriptors representing rigid and reactive behaviour at age 1 year discriminated those in the top 20% of the free salivary cortisol exposure at age 17 years. Genetic association analysis revealed a male-sensitive effect to variation in three specific single nucleotide polymorphisms within selected genes underpinning the overall stress response. Furthermore, support for a polygenic effect on stress-related behaviour in childhood is presented.