In vitro primary cell culture as a physiologically relevant method for preclinical testing of human oncolytic adenovirus

Ad[I/PPT-E1A] is an oncolytic adenovirus that specifically kills prostate cells via restricted replication by a prostate-specific regulatory element. Off-target replication of oncolytic adenoviruses would have serious clinical consequences. As a proposed ex vivo test, we describe the assessment of the specificity of Ad[I/PPT-E1A] viral cytotoxicity and replication in human nonprostate primary cells. Four primary nonprostate cell types were selected to mimic the effects of potential in vivo exposure to Ad[I/PPT-E1A] virus: bronchial epithelial cells, urothelial cells, vascular endothelial cells, and hepatocytes. Primary cells were analyzed for Ad[I/PPT-E1A] viral cytotoxicity in MTS assays, and viral replication was determined by hexon titer immunostaining assays to quantify viral hexon protein. The results revealed that at an extreme multiplicity of infection of 500, unlikely to be achieved in vivo, Ad[I/PPT-E1A] virus showed no significant cytotoxic effects in the nonprostate primary cell types apart from the hepatocytes. Transmission electron microscopy studies revealed high levels of Ad[I/PPT-E1A] sequestered in the cytoplasm of these cells. Adenoviral green fluorescent protein reporter studies showed no evidence for nuclear localization, suggesting that the cytotoxic effects of Ad[I/PPT-E1A] in human primary hepatocytes are related to viral sequestration. Also, hepatocytes had increased amounts of coxsackie adenovirus receptor surface protein. Active viral replication was only observed in the permissive primary prostate cells and LNCaP prostate cell line, and was not evident in any of the other nonprostate cells types tested, confirming the specificity of Ad[I/PPT-E1A]. Thus, using a relevant panel of primary human cells provides a convenient and alternative preclinical assay for examining the specificity of conditionally replicating oncolytic adenoviruses in vivo.

Gene expression profile of primary prostate epithelial and stromal cells in response to sulforaphane or iberin exposure

BACKGROUND: Broccoli consumption has been associated with a reduced risk of prostate cancer. Isothiocyanates (ITCs) derived from glucosinolates that accumulate in broccoli are dietary compounds that may mediate these health effects. Sulforaphane (SF, 4-methylsulphinylbutyl ITC) derives from heading broccoli (calabrese) and iberin (IB, 3-methylsulphinypropyl ITC) from sprouting broccoli. While there are many studies regarding the biological activity of SF, mainly undertaken with cancerous cells, there are few studies associated with IB. METHODS: Primary epithelial and stromal cells were derived from benign prostatic hyperplasia tissue. Affymetrix U133 Plus 2.0 whole genome arrays were used to compare global gene expression between these cells, and to quantify changes in gene expression following exposure to physiologically appropriate concentrations of SF and IB. Ontology and pathway analyses were used to interpret results. Changes in expression of a subset of genes were confirmed by real-time RT-PCR. RESULTS: Global gene expression profiling identified epithelial and stromal-specific gene expression profiles. SF induced more changes in epithelial cells, whereas IB was more effective in stromal cells. Although IB and SF induced different changes in gene expression in both epithelial and stromal cells, these were associated with similar pathways, such as cell cycle and detoxification. Both ITCs increased expression of PLAGL1, a tumor suppressor gene, in stromal cells and suppressed expression of the putative tumor promoting genes IFITM1, CSPG2, and VIM in epithelial cells. CONCLUSION: These data suggest that IB and SF both alter genes associated with cancer prevention, and IB should be investigated further as a potential chemopreventative agent.

Differentiating first episode substance induced and primary psychotic disorders with concurrent substance use in young people

Objective: Substance use is common in first-episode psychosis, and complicates the accurate diagnosis and treatment of the disorder. The differentiation of substance-induced psychotic disorders (SIPD) from primary psychotic disorders (PPD) is particularly challenging. This cross-sectional study compares the clinical, substance use and functional characteristics of substance using first episode psychosis patients diagnosed with a SIPD and PPD. Method: Participants were 61 young people (15-24 years) admitted to a psychiatric inpatient service with first episode psychosis, reporting substance use in the past month. Diagnosis was determined using the Psychiatric Research Interview for DSM-IV Substance and Mental disorders (PRISM-IV). Measures of clinical (severity of psychotic symptoms, level of insight, history of trauma), substance use (frequency/quantity, severity) and social and occupational functioning were also administered. Results: The PRISM-IV differentially diagnosed 56% of first episode patients with a SIPD and 44% with a PPD. Those with a SIPD had higher rates of substance use and disorders, higher levels of insight, were more likely to have a forensic and trauma history and had more severe hostility and anxious symptoms than those with a PPD. Logistic regression analysis indicated a family history of psychosis, trauma history and current cannabis dependence were the strongest predictors of a SIPD. Almost 80% of diagnostic predictions of a SIPD were accurate using this model. Conclusions: This clinical profile of SIPD could help to facilitate the accurate diagnosis and treatment of SIPD versus PPD in young people with first episode psychosis admitted to an inpatient psychiatric service.

'These are my photos of when I was little' : Locating Media Arts in the Primary School Curriculum

‘Media Arts’ has been included as a fifth area of the Arts for the Australian Curriculum which will become mandatory learning for all Australian children from pre-school to Year Six (Y6) from 2014. The current curriculum design is underpinned by an approach familiar to media educators who combine creative practice and critical response to develop students’ media literacies. Media Arts within the Australian Curriculum will place Australia at the forefront of international efforts to promote media education as an entitlement for all children. Even with this mandated endorsement, however, there remains ongoing debate about where to locate media education in school curricula. Historically, media education in Australia has been approached through diverse curriculum activities at the secondary school level. These include subject English’s critical literacy objectives; vocationally oriented media and technology education or ICTs education; and Arts courses using new media technologies for creativity. In this chapter we consider the possibilities and challenges for Media Arts, specifically for primary school student learning. We draw on empirical evidence from a research project that has trialled a Media Arts curriculum with students attending a primary school in a low socio-economic status and culturally diverse community on the outskirts of Brisbane, Queensland.

Practical insights into curricula integration for primary science

As indicated in a previous Teaching Science article, effective planning for curricula integration requires using standards from two (or more) subject areas (e.g., science and English, science and art or science and mathematics), which also becomes the assessment foci for teaching and learning. Curricula integration of standards into an activity necessitates pedagogical knowledge for developing students’ learning in both subject areas. For science education, the skills and tools for curricula integration include the use of other key learning areas (KLAs). A balance between teacher and student-centred science education programs that draw on democratic processes (e.g., Beane, 1997) can be used to make real-world links to target students’ individual needs. This article presents practical ways to commence thinking about curricula integration towards using Australian curriculum standards.

The Yonder project : literacy and social competency development in primary school students through co-creation of live performance

Since 2007 Kite Arts Education Program (KITE), based at Queensland Performing Arts Centre (QPAC), has been engaged in delivering a series of theatre-based experiences for children in low socio-economic primary schools in Queensland. The artist in residence (AIR) project titled Yonder includes performances developed by the children with the support and leadership of teacher artists from KITE for their community and parents/carers,supported by a peak community cultural institution. In 2009,Queensland Performing Arts Centre partnered with Queensland University of Technology (QUT) Creative Industries Faculty (Drama) to conduct a three-year evaluation of the Yonder project to understand the operational dynamics, artistic outputs and the educational benefits of the project. This paper outlines the research findings for children engaged in the Yonder project in the interrelated areas of literacy development and social competencies. Findings are drawn from six iterations of the project in suburban locations on the edge of Brisbane city and in regional Queensland.

Introducing ICT to a primary school in a developing country : a Fijian experience

One aspect of quality education in the 21st century is the availability of digital resources in schools. Many developing countries need to build this capability – not just in terms of technology but teacher capability as well. One of the ways to achieve such capacity is through knowledge sharing between teachers and educators in developed and developing countries. Over time such collaboration can have a lasting impact on all participants on both sides of the digital divide. This paper reports on how such collaboration can occur. It focuses on the initial stages of a long-term initiative where our primary objective is to develop models, which demonstrate how we (in developed countries) can engage productively and meaningfully with schools in developing countries to build their ICT capacity. As part of this initiative, we introduced laptops and LEGO robotics tool kits to a rural primary school in Fiji. We developed ICT activities that aligned with the curriculum in a number of subjects. In addition, we worked with the teachers over two weeks to build their expertise.

Integrating robotics in primary school activities

This article outlines the integration of robotics in two settings in a primary school. This initiative was part of an Australian Research Council project which was undertaken at this school. The article highlights how robotics was integrated in a technology unit in a year four class. It also explains how it was embedded into an after-school program which catered for students from years five to seven. From these experiences further possibilities of engaging with robotics are also discussed.

From vision to reality : views of primary school principals on inclusive education in New South Wales, Australia

This paper discusses the findings of a research study that used semi-structured interviews to explore the views of primary school principals on inclusive education in New South Wales, Australia. Content analysis of the transcript data indicates that principals’ attitudes towards inclusive education and their success in engineering inclusive practices within their school are significantly affected by their own conception of what “inclusion” means, as well as the characteristics of the school community, and the attitudes and capacity of staff. In what follows, we present two parallel conversations that arose from the interview data to illustrate the main conceptual divisions existing between our participants’ conceptions of inclusion. First, we discuss the act of “being inclusive” which was perceived mainly as an issue of culture and pedagogy. Second, we consider the mechanics of “including,” which reflected a more instrumentalist position based on perceptions of individual student deficit, the level of support they may require and the amount of funding they can attract.

Problem solving in the primary school (K-2)

This article focuses on problem solving activities in a first grade classroom in a typical small community and school in Indiana. But, the teacher and the activities in this class were not at all typical of what goes on in most comparable classrooms; and, the issues that will be addressed are relevant and important for students from kindergarten through college. Can children really solve problems that involve concepts (or skills) that they have not yet been taught? Can children really create important mathematical concepts on their own – without a lot of guidance from teachers? What is the relationship between problem solving abilities and the mastery of skills that are widely regarded as being “prerequisites” to such tasks?Can primary school children (whose toolkits of skills are limited) engage productively in authentic simulations of “real life” problem solving situations? Can three-person teams of primary school children really work together collaboratively, and remain intensely engaged, on problem solving activities that require more than an hour to complete? Are the kinds of learning and problem solving experiences that are recommended (for example) in the USA’s Common Core State Curriculum Standards really representative of the kind that even young children encounter beyond school in the 21st century? … This article offers an existence proof showing why our answers to these questions are: Yes. Yes. Yes. Yes. Yes. Yes. And: No. … Even though the evidence we present is only intended to demonstrate what’s possible, not what’s likely to occur under any circumstances, there is no reason to expect that the things that our children accomplished could not be accomplished by average ability children in other schools and classrooms.

Thai family support (TFS) : working with families of young substance users in primary health care

Living with substance users negatively impacts upon family members in many ways, and distress is common. Despite these deep and wide-ranging impacts, supportive interventions for family members in their own right are rarely available. Thailand has substantial and growing problems with substance use, and there is very little support or family members of drug users, especially in community setting. The Thai Family Support (TFS) program was designed for implementation in primary health care units (PCUs) in Thailand. TFS was based on two approaches with existing empirical support in Western contexts—the 5-step method and CRAFT—with adaptations to a Thai setting that included integration with Buddhist practices. Its aims were to increase well-being of family members, reduce mental distress, improve family relationships between family members, and engage substance users in behaviour change. A small-scale randomised controlled trial on TFS with a Delayed Treatment control was conducted, with assessments at 8 weeks (Post 1) and 20-24 weeks (Post 2). Structured interviews with participants and PCU staff and an examination of five case studies augmented the quantitative results. Mixed Model Analyses were applied to quantitative outcomes, and thematic analysis was used for qualitative data. Thirty-six participants (18 in each of Immediate and Delayed Conditions) were recruited. A significant difference at Baseline between the two conditions was observed on the Thai GHQ-28 and Gender, but it was not possible to statistically control for these effects. There was a significant Time by Condition interaction on the Thai GHQ-28, WHOQOL-BREF-THAI and FAS, reflecting greater improvements in the Immediate condition by Post 1, but with the Delayed condition meeting or exceeding that effect by Post 2. On FES Cohesion and Conflict, there were falls across conditions at Post 2, but only Cohesion also showed a Time by Condition interaction, and that effect was consistent with a delayed impact of treatment. Overall, TFS by PCU staff in the Delayed Condition gave similar results to TFS conducted by the researcher, supporting the viability of its dissemination to standard health services. Qualitative data also confirmed the quantitative results. Most participants reported physiological and psychological improvements even though their substance-using relative did not change their drug use behaviour. After completing TFS, participants reported increased knowledge, group support and sharing feeling, having positive patient-professional relationship, having greater knowledge of substance abuse and social support. In particular, they changed their behaviour towards the substance user, resulting in improvements to family relationships. PCU staff gave similar responses on the efficacy of TFS, and saw it as feasible for routine use, although some implementation challenges were identified. The cultural adaptation and in particular the religious activities, were recognised by participants and PCU staff as an important component of TFS to support psychological health and well-being. Findings from this study showed the impact of substance use on family members and difficulties that they experienced when living with the substance users, resulting distresses and burden that may develop severe mental health disease. Drug use policies should be modified to support family members and response to their needs effectively for early prevention. This study also gave preliminary support for application of the TFS program in rural primary care settings and identified some policies that will be required for it to be disseminated more broadly.

Collaborative research into the affordances of place for primary school children’s literacy learning

In the context of culturally diverse high poverty areas of Australia, we have conducted collaborative research with teachers and students in a primary school for more than a decade. Teachers have been exploring the affordances of place‐based pedagogies (Gruenewald & Smith, 2008) for the development of students’ spatial literacies and their understandings of the politics of places and built environments (Comber, Nixon, Ashmore, Loo & Cook, 2006; Comber, Thomson and Wells, 2001). This paper reports on a project in which the affordances of placedbased pedagogy are being explored through teacher inquiries and classroom‐based design experiments (Cobb, Confrey, di Sessa, Lehrer & Schauble, 2003). Located within a large‐scale urban renewal project in which houses are being demolished and families relocated, the original school has been replaced by a larger school that serves a population from a wider area. In this paper we draw on the study to consider the challenges of working with teachers and primary school students to study innovative ideas and practices in educational research. Specifically we consider issues raised by collaborative studies of the affordances of cross curricular projects focusing on social and environmental change to engage students in academic learning and expand their literate repertoires in a changing policy climate.

Preliminary evaluation of a primary care intervention for cry-fuss behaviours in the first 3-4 months of life (‘The Possums Approach’) : effects on cry-fuss behaviours and maternal mood

Problem crying in the first few months of life is both common and complex, arising out of multiple interacting and co-evolving factors. Parents whose babies cry and fuss a lot receive conflicting advice as they seek help from multiple health providers and emergency departments, and may be admitted into tertiary residential services. Conflicting advice is costly, and arises out of discipline-specific interpretations of evidence. An integrated, interdisciplinary primary care intervention (‘The Possums Approach’) for cry-fuss problems in the first months of life was developed from available peer-reviewed evidence. This study reports on preliminary evaluation of delivery of the intervention. A total of 20 mothers who had crying babies under 16 weeks of age (average age 6.15 weeks) completed questionnaires, including the Crying Patterns Questionnaire and the Edinburgh Postnatal Depression Scale, before and 3-4 weeks after their first consultation with trained primary care practitioners. Preliminary evaluation is promising. The Crying Patterns Questionnaire showed a significant decrease in crying and fussing duration, by 1 h in the evening (P = 0.001) and 30 min at night (P = 0.009). The median total amount of crying and fussing in a 24-h period was reduced from 6.12 to 3 h. The Edinburgh Postnatal Depression Scale showed a significant improvement in depressive symptoms, with the median score decreasing from 11 to 6 (P = 0.005). These findings are corroborated by an analysis of results for the subset of 16 participants whose babies were under 12 weeks of age (average age 4.71 weeks). These preliminary results demonstrate significantly decreased infant crying in the evening and during the night and improved maternal mood, validating an innovative interdisciplinary clinical intervention for cry-fuss problems in the first few months of life. This intervention, delivered by trained health professionals, has the potential to mitigate the costly problem of health professionals giving discipline-specific and conflicting advice post-birth.

An ensemble method for predicting subnuclear localizations from primary protein structures

Background Predicting protein subnuclear localization is a challenging problem. Some previous works based on non-sequence information including Gene Ontology annotations and kernel fusion have respective limitations. The aim of this work is twofold: one is to propose a novel individual feature extraction method; another is to develop an ensemble method to improve prediction performance using comprehensive information represented in the form of high dimensional feature vector obtained by 11 feature extraction methods. Methodology/Principal Findings A novel two-stage multiclass support vector machine is proposed to predict protein subnuclear localizations. It only considers those feature extraction methods based on amino acid classifications and physicochemical properties. In order to speed up our system, an automatic search method for the kernel parameter is used. The prediction performance of our method is evaluated on four datasets: Lei dataset, multi-localization dataset, SNL9 dataset and a new independent dataset. The overall accuracy of prediction for 6 localizations on Lei dataset is 75.2% and that for 9 localizations on SNL9 dataset is 72.1% in the leave-one-out cross validation, 71.7% for the multi-localization dataset and 69.8% for the new independent dataset, respectively. Comparisons with those existing methods show that our method performs better for both single-localization and multi-localization proteins and achieves more balanced sensitivities and specificities on large-size and small-size subcellular localizations. The overall accuracy improvements are 4.0% and 4.7% for single-localization proteins and 6.5% for multi-localization proteins. The reliability and stability of our classification model are further confirmed by permutation analysis. Conclusions It can be concluded that our method is effective and valuable for predicting protein subnuclear localizations. A web server has been designed to implement the proposed method. It is freely available at http://bioinformatics.awowshop.com/snlpr​ed_page.php.