Cholesterol enhances phospholipid-dependent activated protein C anticoagulant activity

The influence of cholesterol on activated protein C (APC) anticoagulant activity in plasma and on factor Va inactivation was investigated. Anticoagulant and procoagulant activities of phosphatidylcholine/phosphatidylserine (PC/PS) vesicles containing cholesterol were assessed in the presence and absence of APC using factor Xa-1-stage clotting and factor Va inactivation assays. Cholesterol at approximate physiological membrane levels (30%) in PC/PS (60%/10% w/w) vesicles prolonged the factor Xa-1-stage clotting time dose-dependently in the presence of APC but not in the absence of APC. APC-mediated cleavage of purified recombinant factor Va variants that were modified at specific APC cleavage sites (Q306/Q679-factor Va; Q506/Q679-factor Va) was studied to define the effects of cholesterol on APC cleavage at R506 and R306. When compared to control PC/PS vesicles, cholesterol in PC/PS vesicles enhanced factor Va inactivation and the rate of APC cleavage at both R506 and R306. Cholesterol also enhanced APC cleavage rates at R306 in the presence of the APC cofactor, protein S. In summary, APC anticoagulant activity in plasma and factor Va inactivation as a result of cleavages at R506 and R306 by APC is markedly enhanced by cholesterol in phospholipid vesicles. These results suggest that cholesterol in a membrane surface may selectively enhance APC activities. © 2005 International Society on Thrombosis and Haemostasis.

Building support for learning within a Doctor of Education programme

Professional doctorates were introduced in the 1990s for practitioners to research ‘real-world’ problems relevant to their respective workplace communities and contexts. An array of difficulties faces professional doctoral students as they transition from professionals to practitioner researchers. This study sought to understand the learning journey of a cohort of students at an Australian university and to assess whether the cohort approach provided the necessary support for students to reach their scholarly destinations. Throughout the first 18 months of the programme, focus group interviews and surveys were conducted to gauge students’ experiences and to evaluate developments for support within the programme. Utilising a socio-cultural perspective helped identify and explain the importance of shared practice in fostering learning, the development of academic and researcher identities, and the role of communities of practice. Challenges of managing time and overcoming the professional and academe divide were facilitated by the evolving developments of the programme.

Self-authorship in child care student teachers

Research related to personal epistemology in teacher education indicates that teachers’ beliefs about knowing and learning influence their pedagogical practices. In the current study, we interviewed 31 child care students to investigate the relationship between personal epistemology and beliefs about children’s learning as they engaged in teaching practices with young children. We drew on self authorship theory to analyze this data, which considers the evolving capacity of learners to analyze and make informed judgments about knowledge (personal epistemology)in the light of their professional identity (intrapersonal beliefs) and interdependent social relationships (interpersonal beliefs). The majority of students described practical personal epistemologies which involved either modeling, reflection on, or evaluation of practical strategies. These epistemologies have implications for child care teachers’ professional identities and their relationships with families, children, and staff in child care contexts.

Teachers' personal epistemologies and teacher education : emergent themes and future research

This chapter summarizes the responses to four questions in each of the chapters in this volume. The questions addressed the use of a conceptual framework that guides the chapter, issues of domain-generality, how personal epistemology relates to teaching, and how personal epistemologies change. We concluded that all of the chapters discussed the distinction between constructivist and transmission teaching practices, while suggesting that there are many inconsistencies in understanding the relationship between the nature of beliefs and teachers’ practices regardless of the relative sophistication of teachers’ personal epistemologies. We also summarized a multi-component instructional model for calibrating teaching practices based on suggestions in each of the chapters, and made four suggestions for future research, including the need for an integrated theory that accounts for the development and manifestations of personal pistemology in the classroom, the generalizability of fi ndings across different measurements, a set of guidelines to promote teacher epistemological change, and an explicit instructional model that explains the development and calibration of beliefs and practices. The goal of this volume was to examine the relationship between teachers’ personal epistemologies and teacher education. Sixteen different chapters addressed one or more aspects of this issue. Although each of the chapters addressed different aspects of teachers’ personal epistemologies, a number of common themes are apparent across the chapters. We believe it is useful to articulate these themes in greater detail to provide a better retrospective understanding of this volume, as well as a better prospective framework for future research and changes to teacher training programs. We divide this chapter into two main sections. The fi rst section addresses four key questions about the nature of teachers’ personal epistemologies that were discussed in the introductory chapter as part of a larger set of questions. These questions focus on how to conceptualize these beliefs as explicit models; whether beliefs are domain-specifi c or domain-general; how beliefs are related to teaching; and how beliefs change over time. We provide a summary of each chapter in terms of these four questions. The second section proposes four general suggestions for future research based on the studies reported within this volume.

A comparison of the effects of a chlamydial vaccine administered during or after a C. muridarum urogenital infection of female mice

There are approximately 92 million new chlamydial infections of the genital tract in humans diagnosed each year, costing health care systems billions of dollars in treatment not only of acute infections, but also of associated inflammatory sequelae, such as pelvic inflammatory disease (PID) and ectopic pregnancy. These numbers are increasing at a steady rate and, due to the asymptomatic nature of infections, the incidence may be underestimated and the costs of treatment therefore higher. Over the previous few decades there has been a large amount of research into the development of an efficacious vaccine against genital tract chlamydial infections. The majority of this research has focused on females, due to the high rate of development of associated diseases, including PID, which can lead to ectopic pregnancy and infertility. In light of the increasing infection rates that have occurred despite the availability of antibiotics, and the asymptomatic nature of chlamydial infections, it is imperative that an efficacious vaccine that protects against infection and associated pathology be developed.

Gonadotropin-induced ovarian cancer cell migration and proliferation require extracellular signal-regulated kinase 1/2 activation regulated by calcium and protein kinase Cδ

The gonadotropin hypothesis proposes that elevated serum gonadotropin levels may increase the risk of epithelial ovarian cancer (EOC). We have studied the effect of treating EOC cell lines (OV207 and OVCAR-3) with FSH or LH. Both gonadotropins activated the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase 1/2 (ERK1/2) pathway and increased cell migration that was inhibited by the MAPK 1 inhibitor PD98059. Both extra- and intracellular calcium ion signalling were implicated in gonadotropin-induced ERK1/2 activation as treatment with either the calcium chelator EGTA or an inhibitor of intracellular calcium release, dantrolene, inhibited gonadotropin-induced ERK1/2 activation. Verapamil was also inhibitory, indicating that gonadotropins activate calcium influx via L-type voltage-dependent calcium channels. The cAMP/protein kinase A (PKA) pathway was not involved in the mediation of gonadotropin action in these cells as gonadotropins did not increase intracellular cAMP formation and inhibition of PKA did not affect gonadotropin-induced phosphorylation of ERK1/2. Activation of ERK1/2 was inhibited by the protein kinase C (PKC) inhibitor GF 109203X as well as by the PKCδ inhibitor rottlerin, and downregulation of PKCδ was inhibited by small interfering RNA (siRNA), highlighting the importance of PKCδ in the gonadotropin signalling cascade. Furthermore, in addition to inhibition by PD98059, gonadotropin-induced ovarian cancer cell migration was also inhibited by verapamil, GF 109203X and rottlerin. Similarly, gonadotropin-induced proliferation was inhibited by PD98059, verapamil, GF 109203X and PKCδ siRNA. Taken together, these results demonstrate that gonadotropins induce both ovarian cancer cell migration and proliferation by activation of ERK1/2 signalling in a calcium- and PKCδ-dependent manner.

Grading : harmonising standards and stakeholder expectations

This paper suggests that when a course is planned within one culture for delivery to members of another culture, appropriate quality control of assessment becomes an issue of major proportions. Based on their experience of presenting an Aid Agency-funded Masters course in a developing country in the Pacific, the authors describe the processes to address the needs and wants of all the stakeholders, with different cultural expectations. Maintaining a balance between domestic and Pacific student cohorts regarding resources and opportunities for study was especially challenging. However, grounding grades in course curriculum and clearly stated objectives permitted the teaching team to meet external requirements while maintaining their professional and academic freedom.

Microarray expression profiling in melanoma reveals a BRAF mutation signature

We have used microarray gene expression profiling and machine learning to predict the presence of BRAF mutations in a panel of 61 melanoma cell lines. The BRAF gene was found to be mutated in 42 samples (69%) and intragenic mutations of the NRAS gene were detected in seven samples (11%). No cell line carried mutations of both genes. Using support vector machines, we have built a classifier that differentiates between melanoma cell lines based on BRAF mutation status. As few as 83 genes are able to discriminate between BRAF mutant and BRAF wild-type samples with clear separation observed using hierarchical clustering. Multidimensional scaling was used to visualize the relationship between a BRAF mutation signature and that of a generalized mitogen-activated protein kinase (MAPK) activation (either BRAF or NRAS mutation) in the context of the discriminating gene list. We observed that samples carrying NRAS mutations lie somewhere between those with or without BRAF mutations. These observations suggest that there are gene-specific mutation signals in addition to a common MAPK activation that result from the pleiotropic effects of either BRAF or NRAS on other signaling pathways, leading to measurably different transcriptional changes.