Minimal mutation of the cytoplasmic tail inhibits the ability of E-cadherin to activate Rac but not phosphatidylinositol 3-kinase - Direct evidence of a role for cadherin-activated Rac signaling in adhesion and contact formation

Classic cadherins are adhesion-activated cell signaling receptors. In particular, homophilic cadherin ligation can directly activate Rho family GTPases and phosphatidylinositol 3-kinase (PI3-kinase), signaling molecules with the capacity to support the morphogenetic effects of these adhesion molecules during development and disease. However, the molecular basis for cadherin signaling has not been elucidated, nor is its precise contribution to cadherin function yet understood. One attractive hypothesis is that cadherin-activated signaling participates in stabilizing adhesive contacts ( Yap, A. S., and Kovacs, E. M. ( 2003) J. Cell Biol. 160, 11-16). We now report that minimal mutation of the cadherin cytoplasmic tail to uncouple binding of p120-ctn ablated the ability of E-cadherin to activate Rac. This was accompanied by profound defects in the capacity of cells to establish stable adhesive contacts, defects that were rescued by sustained Rac signaling. These data provide direct evidence for a role of cadherin-activated Rac signaling in contact formation and adhesive stabilization. In contrast, cadherin-activated PI3-kinase signaling was not affected by loss of p120-ctn binding. The molecular requirements for E-cadherin to activate Rac signaling thus appear distinct from those that stimulate PI3-kinase, and we postulate that p120-ctn may play a central role in the E-cadherin-Rac signaling pathway.

Changing student teachers' attitudes towards disability and inclusion

A total of 274 preservice teacher education students were surveyed at the beginning and end of a one-semester unit on Human Development and Education which combined formal instruction with structured fieldwork experiences. The latter included interviewing community members regarding their knowledge of Down syndrome and opinions on inclusive education, and writing an associated report. At the end of semester, not only had student teachers acquired more accurate knowledge of Down syndrome, together with more positive attitudes towards the inclusive education of children with Down syndrome, but their attitudes towards disability in general had also changed, and they reported greater ease when interacting with people with disabilities. The study illustrated the value of combining information-based instruction with structured fieldwork experiences in changing attitudes towards disability and inclusion. It also demonstrated that raising awareness of one disability may lead to changes in attitudes towards disability in general.

Partnerships in mental health: Addressing barriers to social inclusion

Current trends in mental health services emphasize working in partnership with consumers and other government and non-government community organizations for improvement in quality of life for service users. People with a mental illness experience social exclusion, thereby limiting their ability to participate fully in community life. Occupational therapists have a substantial role to play in helping service users to overcome barriers to their community inclusion. Partnerships need to be formed to increase access to community resources and participation in activities that are enjoyed by other members of the community. Such partnerships have a health promotion emphasis and foster the relationship between mental-health services and the wider community, thus shifting the focus from direct occupational therapy service delivery to community-based rehabilitation interventions.This article describes the development, implementation and evaluation of an Australian healthy lifestyle course devised to meet the identified rehabilitation goals of people with mental illness to lose weight, get fit, commence vocational study and get a job. The course was run in partnership between mental health consumers, occupational therapists from the St George Mental Health Rehabilitation Service and staff and students from the Sutherland College of Technical and Further Education.

Relation between cell disruption conditions, cell debris particle size, and inclusion body release

The efficiency of physical separation of inclusion bodies from cell debris is related to cell debris size and inclusion body release and both factors should be taken into account when designing a process. In this work, cell disruption by enzymatic treatment with lysozyme and cellulase, by homogenization, and by homogenization with ammonia pretreatment is discussed. These disruption methods are compared on the basis of inclusion body release, operating costs, and cell debris particle size. The latter was measured with cumulative sedimentation analysis in combination with membrane-associated protein quantification by SDS-PAGE and a spectrophotometric pepticloglycan quantification method. Comparison of the results obtained with these two cell debris quantification methods shows that enzymatic treatment yields cell debris particles with varying chemical composition, while this is not the case with the other disruption methods that were investigated. Furthermore, the experiments show that ammonia pretreatment with homogenization increases inclusion body release compared to homogenization without pretreatment and that this pretreatment may be used to control the cell debris size to some extent. The enzymatic disruption process gives a higher product release than homogenization with or without ammonia pretreatment at lower operating costs, but it also yields a much smaller cell debris size than the other disruption process. This is unfavorable for centrifugal inclusion body purification in this case, where cell debris is the component going to the sediment and the inclusion body is the floating component. Nevertheless, calculations show that centrifugal separation of inclusion bodies from the enzymatically treated cells gives a high inclusion body yield and purity. (C) 2004 Wiley Periodicals, Inc.

Wolbachia-induced cytoplasmic incompatibility as a means for insect pest population control

Biological control is the purposeful introduction of parasites, predators, and pathogens to reduce or suppress pest populations. Wolbachia are inherited bacteria of arthropods that have recently attracted attention for their potential as new biocontrol agents. Wolbachia manipulate host reproduction by using several strategies, one of which is cytoplasmic incompatibility (CI) [Stouthamer, R., Breeuwer, J. A. J. & Hurst, G. D. D. (1999) Annu. Rev. Microbiol. 53,71-102]. We established Wolbachia-infected lines of the medfly Ceratitis capitata using the infected cherry fruit fly Rhagoletis cerasi as donor. Wolbachia induced complete CI in the novel host. Laboratory cage populations were completely suppressed by single releases of infected males, suggesting that Wolbachia-induced CI could be used as a novel environmentally friendly tool for the control of medfly populations. The results also encourage the introduction of Wolbachia into pest and vector species of economic and hygenic relevance to suppress or modify natural populations.

Membrane insertion of anthrax protective antigen and cytoplasmic delivery of lethal factor occur at different stages of the endocytic pathway

The protective antigen (PA) of anthrax toxin binds to a cell surface receptor, undergoes heptamerization, and binds the enzymatic subunits, the lethal factor (LF) and the edema factor (EF). The resulting complex is then endocytosed. Via mechanisms that depend on the vacuolar ATPase and require membrane insertion of PA, LF and EF are ultimately delivered to the cytoplasm where their targets reside. Here, we show that membrane insertion of PA already occurs in early endosomes, possibly only in the multivesicular regions, but that subsequent delivery of LF to the cytoplasm occurs preferentially later in the endocytic pathway and relies on the dynamics of internal vesicles of multivesicular late endosomes.

Implications of social capital for the inclusion of people with disabilities and families in community life

This paper seeks to ascertain the usefulness of the theory of social capital as a framework for developing and sustaining the inclusion of people with disabilities and families in community life. We discuss the theoretical elements of social capital and assess its relevance when understanding both the experiences of people with disabilities and their families and the possible implications for policy and programme efforts to promote inclusion. Preliminary findings from two studies of the experiences and social networks of people with disabilities and their families in communities in regional and rural Australia are presented. It is argued that to date, people with disabilities and their families have largely been excluded from the broader social capital debate and that social capital thinking has had minimal influence on efforts to achieve the inclusion of people with disabilities into community life. It is further argued that new paradigms of support are needed that build capacity and social capital through working alongside individuals and families to influence not only outcomes for them, but also for the communities on which they live. The local area coordination model as it has developed in Australia since 1989 provides some instructive signposts for integrating individual, family and community approaches. It is concluded that social capital theory can make a contribution to inclusion theory and practice but we should use it with circumspection.

Social inclusion, new regionalism and associational governance: The Queensland experience

In recent years there has been a resurgence of decentralized social governance concerned with the spatial dimensions of disadvantage. This article examines aspects of this resurgence in the Australian state of Queensland where, after the hasty birth of 'place management' in response to the rise of 'Hansonism', a plethora of 'joined-up' policy initiatives were undertaken in relation to the regional dimensions of poverty. We propose that these trends reflect in part new ways of thinking about the spatial aspects of disadvantage which have emerged in recent years and which have the potential to take regional policy beyond the narrow confines imposed by neoliberal economic orthodoxy. These new ways of thinking have arisen in social policy through the refraining of disadvantage in terms of social exclusion and in regional economic policy through the influence of the so-called 'new regionalism'. The article shows how together these bodies of theory point us towards a new model of 'associational governance'. The article reviews recent Queensland experience and indicates those features of 'associational governance' which have become characteristic of locality-based social policy ideas in Queensland. 'Joined-up' and regional policy aspirations of the Queensland State government have shown the influence of these new approaches. The political and policy sustainability of these trends, however, is uncertain. The lingering shadow of managerialism and neoliberal policy frameworks remains a significant barrier to the innovation and viability of these approaches. More directly, the inherent limits of the 'local' or 'regional' initiatives in the face of broader national and global factors will significantly constrain the capacity of associational governance systems to deliver positive democratic, social and economic outcomes. The article examines recent Queensland policy refors in light of this complex set of factors and concludes by offering directions for future research and policy development.

Genetic variation for carbon isotope discrimination in sunflower: Association with transpiration efficiency and evidence for cytoplasmic inheritance

Plants accumulate isotopes of carbon at different rates because of discrimination against C-13 relative to C-12. In plants that fix carbon by the C-3 pathway, the amount of discrimination correlates negatively with transpiration efficiency (TE) where TE is the amount of dry matter accumulated per unit water transpired. Therefore, carbon isotope discrimination (Delta) has become a useful tool for selecting genotypes with improved TE and performance in dry environments. Surveys of 161 sunflower (Helianthus spp.) genotypes of diverse origin revealed a large and unprecedented range of genetic variation for Delta (19.5-23.8parts per thousand). A strong negative genetic correlation (r(g)) between TE and Delta (r(g) = -0.87, P < 0.001) was observed in glasshouse studies. Gas exchange measurements of field grown plants indicated that Delta was strongly correlated with stomatal conductance to water vapor (g), (r(g) 0.64, P < 0.01), and the ratio of net assimilation rate (A) to g, (r(g) = 0.86, P < 0.001), an instantaneous measure of TE. Genotype CMSHA89MAX1 had the lowest TE (and highest Delta) of all genotypes tested in these studies and low yields in hybrid combination. Backcrossing studies showed that the TE of this genotype was due to an adverse effect of the MAX1 cytoplasm, which was inherited from the diploid perennial H. maximiliani Schrader. Overall, these studies suggested that there is an excellent opportunity for breeders to develop sunflower germplasm with improved TE. This can be achieved, in part, by avoiding cytoplasms such as the MAX1 cytoplasm.

Heterozygote effects in mice with partial truncations in the growth hormone receptor cytoplasmic domain: Assessment of growth parameters and phenotype

The GH receptor (GHR) is essential for normal postnatal growth and development, and the molecular basis of GHR action has been studied intensively. Clinical case studies and more recently mouse models have revealed the extensive phenotype of impaired GH action. We recently reported two new mouse models, possessing cytoplasmic truncations at position 569 (plus Y539/545-F) and 391, which were created to identify functional subdomains within the cytoplasmic signaling domain. In the homozygous state, these animals show progressively impaired postnatal growth coupled with complex changes in gene expression. We describe here an extended phenotype analysis encompassing the heterozygote state to identify whether single copies of these mutant receptors bring about partial or dominant-negative phenotypes. It appears that the retention of the ubiquitin-dependent endocytosis motif the N-terminal cytoplasmic domain permits turnover of these mutant receptors because no dominant-negative phenotype is seen. Nonetheless, we do observe partial impairment of postnatal growth in heterozygotes supporting limited haploinsufficiency. Reproductive function is impaired in these models in a progressive manner, in parallel with loss of signal transducer and activator of transcription-5 activation ability. In summary, we describe a more comprehensive phenotypic analysis of these mouse models, encompassing overall and longitudinal body growth, reproductive function, and hormonal status in both the heterozygote and homozygote state. Our results suggest that patients expressing single copies of similarly mutated GHRs would not display an obvious clinical phenotype.