Development of an oligonucleotide-based SNP detection method on lateral flow strips using hexapet tages

Detection of point mutations or single nucleotide polymorphisms (SNPs) is important in relation to disease susceptibility or detection in pathogens of mutations determining drug resistance or host range. There is an emergent need for rapid detection methods amenable to point-of-care applications. The purpose of this study was to reduce to practice a novel method for SNP detection and to demonstrate that this technology can be used downstream of nucleic acid amplification. The authors used a model system to develop an oligonucleotide-based SNP detection system on nitrocellulose lateral flow strips. To optimize the assay they used cloned sequences of the herpes simplex virus-1 (HSV-1) DNA polymerase gene into which they introduced a point mutation. The assay system uses chimeric polymerase chain reaction (PCR) primers that incorporate hexameric repeat tags ("hexapet tags"). The chimeric sequences allow capture of amplified products to predefined positions on a lateral flow strip. These "hexapet" sequences have minimal cross-reactivity and allow specific hybridization-based capture of the PCR products at room temperature onto lateral flow strips that have been striped with complementary hexapet tags. The allele-specific amplification was carried out with both mutant and wild-type primer sets present in the PCR mix ("competitive" format). The resulting PCR products carried a hexapet tag that corresponded with either a wild-type or mutant sequence. The lateral flow strips are dropped into the PCR reaction tube, and mutant sequence and wild-type sequences diffuse along the strip and are captured at the corresponding position on the strip. A red line indicative of a positive reaction is visible after 1 minute. Unlike other systems that require separate reactions and strips for each target sequence, this system allows multiplex PCR reactions and multiplex detection on a single strip or other suitable substrates. Unambiguous visual discrimination of a point mutation under room temperature hybridization conditions was achieved with this model system in 10 minutes after PCR. The authors have developed a capture-based hybridization method for the detection and discrimination of HSV-1 DNA polymerase genes that contain a single nucleotide change. It has been demonstrated that the hexapet oligonucleotides can be adapted for hybridization on the lateral flow strip platform for discrimination of SNPs. This is the first step in demonstrating SNP detection on lateral flow using the hexapet oligonucleotide capture system. It is anticipated that this novel system can be widely used in point-of-care settings.

Retinal serotonin, eye growth and myopia development in chick

Myopia (short-sightedness) is a visual problem associated with excessive eye growth and vitreous chamber expansion. Within the eye serotonin (5-hydroxytryptamine, 5-HT) appears to have a variety of effects, it alters retinal amacrine cell processing, increases intraocular pressure, constricts ocular blood vessels, and is also mitogenic. This study sought to determine the role of the retinal serotonin system in eye growth regulation. Myopia was produced in 7-day-old chicks using -15 D spectacle lenses (LIM) and form deprivation (FDM). The effect on LIM and FDM of daily intravitreal injections of a combination of 5-HT receptor antagonists (1, 10, 50 mu M), 5-HT2 selective antagonist (Mianserin 0.5, 20 mu M) were assessed. Counts were performed of serotonin and tyrosine hydroxylase positive neurons and the relative density used to account for areal changes due to eye growth. The effect of LIM and lens-induced hyperopia (LIH) on the numbers of 5-HT-containing amacrine cells in the retina were then determined. The combination of the 5-HT receptor antagonists inhibited LIM by approximately half (1 mu M RE: -7.12 +/- 1.0 D, AL: 0.38 +/- 0.06 mm vs. saline RE: -13.19 +/- 0.65 D, AL: 0.64 +/- 0.03 mm. RE: p < 0.01, AL: p < 0.01), whereas FDM was not affected (1 mu M RE: -8.88 +/- 1.10 D). These data suggest that serotonin has a stimulatory role in LIM, although high doses of serotonin were inhibitory (1 mu M RE: -9.30 +/- 1.34 D). 5-HT immunoreactivity was localised to a subset of amacrine cell bodies in the inner nuclear layer of the retina, and to two synaptic strata in the inner plexiform layer. LIM eyes had increased numbers of 5-HT-containing amacrine cells in the central retina (12.5%). Collectively, these results suggest that manipulations to the serotonin system can alter the eye growth process but the role of the transmitter system within this process remains unclear. (c) 2005 Elsevier Ltd. All rights reserved.

Post-hatching growth and development of the pectoral and pelvic limbs in the black noddy, Anous minutus

The black tern (Anous minutus) uses a semi-precocial growth strategy. Terrestrial locomotor capacity occurs soon after hatching, but pectoral limb development is delayed and flight is not possible until about post-hatching day 50. A growth series (hatchlings to fledglings) was used to explore how limb musculoskeletal development varied with body mass. In the pelvic limb, bone lengths scaled isometrically or with negative allometry. Gastrocnemius muscle mass and the failure load and stiffness of the tibiotarsus scaled isometrically. In the pectoral limb, pectoralis and supracoracoideus muscle masses increased with strong positive allometry that was mirrored by increases in wing bone strength and stiffness. Bending strength (σult) and modulus (E) remained fairly constant throughout development to fledging for all limb bones. The moment of inertia (I) scaled with negative allometry for the tibiotarsus and with strong positive allometry in the wing bones. Differences in σult and E of the tibiotarsus between pre-fledged chicks and adults was due, primarily, to increases in bone density rather than increases in the moment of inertia of the skeletal elements, whereas σult of wing bones was a function of increases in both bone density and I. Early development of functional pelvic limbs in tree-nesting birds is relatively unusual, and presumably reflects a familial trait that does not appear to compromise breeding success in this species.

Reactions to positive deviance: Social identity and attribution dimensions

This research examines whether evaluations of positive deviates (i.e. high achieving group members) are influenced by the attributions they make for their performance. We argue that ingroup positive deviates who make group attributions help enhance the ingroup's image and thus attract favorable evaluations. In Experiment 1, ingroup positive deviates who made group attributions were generally evaluated more favorably than ingroup positive deviates who made individual attributions. There was also evidence that the positive deviates' attribution style influenced group and self-evaluations. Evaluations of outgroup positive deviates were not influenced by their attribution style. In Experiment 2, an ingroup positive deviate who was successful and attributed that success to the group was upgraded relative to an ingroup positive deviate who made individual attributions. Group evaluations were also higher when the positive deviate made group attributions. This pattern did not emerge when the positive deviate failed. The results are discussed from a social identity perspective.

Introducing evidence-based family assessment and therapy in child and youth mental health services: Applying systemic principles to maintain, sustain and build capacity

The purpose of this article is to overview the context of the mental health service in which we work, and family therapy's status prior to and after the impact of changes wrought by the introduction of the National Mental Health Policy. We then explore some key issues that we think contribute to the persistence of the occlusion of family therapy in child psychiatric services; and the strategies that we developed and are continuing to develop to support change, finally, we describe the use of a family assessment instrument that we believe is central to our change strategy.

Does the Triple P-Positive Parenting Program provide value for money?

Objective: The aim of the present study was to investigate the economic case for the implementation of the Triple P- Positive Parenting Program on a population basis in Queensland, Australia, in order to reduce the prevalence of conduct disorder in children. Method: Threshold analysis was undertaken together with a limited cost-effectiveness analysis. Results: The Triple P-Positive Parenting Program is a dominant intervention; that is, it costs less than the amount it saves, until the reduction in prevalence falls below 7% where net costs become positive. Conclusions: Triple P is likely to be a worthwhile use of limited health funds. The economic case is promising, but further research is required to confirm the study results.

Water, Communities and Mineral Resource Development: Understanding the Risks and Opportunities

Issues around water – its quality, use, availability and environmental value – can be a major point of tension between mineral projects and local communities. Failure to manage these issues appropriately can jeopardise the obtaining of regulatory approvals for new projects and place at risk the ‘social licence to operate’ of existing operations. Conversely, there may also be significant opportunities for companies to engage constructively with communities over water issues and to make a positive contribution to the sustainable development of these communities. Using case studies, this paper will explore the various types of social risks and opportunities associated with water and mineral resource development and identify key learnings relating to the management of these issues.

AS INTERFACES DA PROMOÇÃO DA SAÚDE E UM PROGRAMA DE APRENDIZAGEM PROFISSIONAL

O trabalho desenvolvido com adolescentes, pensando na promoção de sua saúde, bem como em seu desenvolvimento saudável por meio de espaços que atendam suas necessidades, tem sido de responsabilidade também de organizações sociais. Em especial, quando se tem o objetivo de prepará- los para o mundo do trabalho, porém com uma proposta mais ampla, ou seja, como uma preparação para suas relações na vida, pela passagem de um processo de aprendizagem, de amadurecimento, por meio da construção de ações significativas para vida. Esta é a proposta do Centro de Orientação ao Adolescente de Campinas COMEC , organização não governamental, que conta com um Progr ama de Aprendizagem Profissional- PAP . Sendo assim, esse estudo tem como objetivo caracterizar, por meio de prontuários, uma amostra de 280 adolescentes, compreender a percepção deles no processo vivido e identificar as expectativas na admissão, demissão e desligamento do programa - PAP. A pesquisa teve como participantes adolescentes de 14 a 17 anos, do gênero feminino e masculino, do ensino fundamental e médio (7ª série E.F até 3ª série E.M). Os formulários foram preenchidos em três momentos: primeiro quando o adolescente participava da seleção para ingressar no programa, segundo, quando ele iniciava o trabalho na empresa e no término do contrato do adolescente com o programa. Os resultados obtidos por meio dos prontuários dos adolescentes mostraram que a percepção que eles têm a respeito do programa é positiva. Perceberam-se sentimentos de medo, insegurança, alegrias e desejos a respeito das atividades que os preparam para a inserção no mercado formal de trabalho, e o quanto o acompanhamento pelo programa permite outro olhar para esta inclusão. Ainda que o trabalho juvenil não seja a solução para as necessidades econômicas de muitas famílias, o trabalho educativo tem sido visto como uma forma de permitir que o adolescente vivencie sua primeira experiência de trabalho, respeitando sua condição de pessoa em desenvolvimento. Porém, em alguns casos, acabam por contribuir de forma significativa no orçamento familiar.(AU)

Positive psychology approaches to public services leadership: An introduction to strengths-based leadership

To readers of the popular press, the words ‘positive psychology’ may conjure up images of happiness gurus and people having their feet massaged, their heads resting peacefully on pink, fluffy clouds. But in this article, our aim is to demonstrate how the new science of positive psychology speaks powerfully to - and has much to contribute to - the development of leadership and the practices and processes of organisations, whether in the public or private sectors. Much of our work is concerned with the applications of this new field, and particularly with building strengths-based organisations. A key pillar of this work is around enabling strengths-based leadership, and provides our focus for this article.

An expert system for the development of a health and safety policy/risk assessment in the plastics industry

Health and safety policies may be regarded as the cornerstone for positive prevention of occupational accidents and diseases. The Health and Safety at Work, etc Act 1974 makes it a legal duty for employers to prepare and revise a written statement of a general policy with respect to the health and safety at work of employees as well as the organisation and arrangements for carrying out that policy. Despite their importance and the legal equipment to prepare them, health and safety policies have been found, in a large number of plastics processing companies (particularly small companies), to be poorly prepared, inadequately implemented and monitored. An important cause of these inadequacies is the lack of necessary health and safety knowledge and expertise to prepare, implement and monitor policies. One possible way of remedying this problem is to investigate the feasibility of using computers to develop expert system programs to simulate the health and safety (HS) experts' task of preparing the policies and assisting companies implement and monitor them. Such programs use artificial intelligence (AI) techniques to solve this sort of problems which are heuristic in nature and require symbolic reasoning. Expert systems have been used successfully in a variety of fields such as medicine and engineering. An important phase in the feasibility of development of such systems is the engineering of knowledge which consists of identifying the knowledge required, eliciting, structuring and representing it in an appropriate computer programming language.

Foreign direct investment and economic development:an interactive relationship

With the increasing importance of Foreign Direct Investment (FDI), there have been substantial studies on this issue, both empirically and theoretically. However, most existing studies focus on either the impacts of FDI presence or the determinants of FDI inflows, ignoring the fact that inward FDI and economic development may simultaneously affect each other. This thesis sets out to examine the interactive effects between FDI and economic development. The whole thesis is composed of five chapters. Chapter One is an overall introduction to the thesis. Chapter Two presents a theoretical study and chapter Two and Three provide two empirical studies. Chapter Five concludes. Chapter Two presents a theoretical two-sector model that features the importance of human capital in attracting foreign investment. This model theoretically explains why FDI is more likely to occur among countries that are similar in terms of human capital and technology. On the other hand, MNCs must train local employees to work with firm-specific technology and hence improve the technological skills of local workers. In Chapter Two, an empirical model is constructed to detect whether the productivities of foreign and local firms impact each other. The model is tested on China’s data at the industry level. The results indicate that productivity growth of local and foreign firms are jointly determined. Evidence also suggests that the extent to which spillovers occur varies with difference technology levels of local firms. Chapter Four investigates the relationship between FDI and economic grown based on a panel of data for 84 countries over the period 1970-1999. Both equations of FDI inflow and GDP growth are examined. The results indicate that FDI not only directly promotes economic growth by itself, but also indirectly does so via its interaction terms. There is a strong positive interaction effect of FDI with human capital and a strong negative interaction effect of FDI with technology gap on economic growth in developing countries.

The impact of brand owner on consumers' brand perceptions : a development of Heider's Balance Theory

Studies have shown that the brand “owner” is very influential in positioning the brand and when the brand “owner” ceases his or her active role the brand will be perceived differently by the consumers. Balance Theory (HBT), a cognitive psychological theory, studies the triadic relationships between two persons and an entity and predicts that when a person’s original perception of the relationship is disturbed, the person restructures to a new balanced perception. Consequently, this research was undertaken to: conceptualize the brand owner’s impact on consumer’s brand perception; test the applicability of both the static and dynamic predictions of the Heider’s Balance Theory in brand owner-consumer-brand relation (OCB); construct and test a model of brand owner-consumer-brand relation; and examine if personality has an influence on OCB. A discovery-oriented approach was taken to understand the selected market segment, the ready-to-wear and diffusion lines of international designer labels. Chinese Brand Personality Scale, fashion proneness and hedonic and utilitarian shopping scales were developed, and validated. 51 customers were surveyed. Both traditional and extended methods used in the Balance Theory were employed in this study. Responses to liked brand have been used to test and develop the model, while those for disliked brand were used for test and confirmation. A “what if’ experimental approach was employed to test the applicability of dynamic HBT theory in OCB Model. The hypothesized OCB Model has been tested and validated. Consumers have been found to have separate views on the brand and the brand owner; and their responses to contrasting ethical and non-ethical news of the brand owner are different. Personality has been found to have an influence and two personality adapted models have been tested and validated. The actual results go beyond the prediction of the Balance Theory. Dominant triple positive balance mode, dominant negative balance mode, and mode of extreme antipathy have been found. It has been found that not all balanced modes are good for the brand. Contrary to Heider’s findings, simply liking may not necessarily lead to unit relation in the OCB Model.