Parasite diversity, patterns of MHC II variation and olfactory based mate choice in diverging three-spined stickleback ecotypes

Ecological speciation has been the subject of intense research in evolutionary biology but the genetic basis of the actual mechanism driving reproductive isolation has rarely been identified. The extreme polymorphism of the major histocompatibility complex (MHC), probably maintained by parasite-mediated selection, has been proposed as a potential driver of population divergence. We performed an integrative field and experimental study using three-spined stickleback river and lake ecotypes. We characterized their parasite load and variation at MHC class II loci. Fish from lakes and rivers harbor contrasting parasite communities and populations possess different MHC allele pools that could be the result of a combined action of genetic drift and parasite-mediated selection. We show that individual MHC class II diversity varies among populations and is lower in river ecotypes. Our results suggest the action of homogenizing selection within habitat type and diverging selection between habitat types. Finally, reproductive isolation was suggested by experimental evidence: in a flow channel design females preferred assortatively the odor of their sympatric male. This demonstrates the role of olfactory cues in maintaining reproductive isolation between diverging fish ecotypes.

Molecular diagnosis for heterogeneous genetic diseases with high-throughput DNA sequencing applied to retinitis pigmentosa

BACKGROUND:
The genetic heterogeneity of many Mendelian disorders, such as retinitis pigmentosa which results from mutations in over 40 genes, is a major obstacle to obtaining a molecular diagnosis in clinical practice. Targeted high-throughput DNA sequencing offers a potential solution and was used to develop a molecular diagnostic screen for patients with retinitis pigmentosa.
METHODS:
A custom sequence capture array was designed to target the coding regions of all known retinitis pigmentosa genes and used to enrich these sequences from DNA samples of five patients. Enriched DNA was subjected to high-throughput sequencing singly or in pools, and sequence variants were identified by alignment of up to 10 million reads per sample to the normal reference sequence. Potential pathogenicity was assessed by functional predictions and frequency in controls.
RESULTS AND CONCLUSIONS:
Known homozygous PDE6B and compound heterozygous CRB1 mutations were detected in two patients. A novel homozygous missense mutation (c.2957A?T; p.N986I) in the cyclic nucleotide gated channel ß1 (CNGB1) gene predicted to have a deleterious effect and absent in 720 control chromosomes was detected in one case in which conventional genetic screening had failed to detect mutations. The detection of known and novel retinitis pigmentosa mutations in this study establishes high-throughput DNA sequencing with DNA pooling as an effective diagnostic tool for heterogeneous genetic diseases.

Confined Site Construction: Issues Regarding Implementation of Health and Safety

The objective of this paper is to identify the various managerial constraints, difficulties and issues encountered and resulting strategies adopted, to aid in the management of the various and often complex health and safety concerns, which occur within a confined construction site. This is achieved through classifying the various managerial burdens encountered with the numerous strategies adopted, to ensure the successful management of such confined environments within the realm of health and safety. Through an extensive literature review and detailed interviews, a comprehensive insight into the health and safety concerns within a confined construction site environment is portrayed. The leading managerial strategies to the management of health and safety on confined construction sites may be listed as follows; (1) Traffic Management Plan, (2) Effective Resource Management Plan, (3) Temporary Facilities Management Plan, (4) Safe System of Work Plan, (5) Site Safety Plan, (6) Design Site Layout, (7) Space Management Plan, (8) Effective Program Management, and (9) Space Scheduling. Based on the research, it can be concluded, that through effective management of these issues identified coupled with implementing the various strategies highlighted; successful management of health and safety within a confined construction site environment is attainable.

Short interfering RNA-mediated knockdown of drosha and pasha in undifferentiated Meloidogyne incognita eggs leads to irregular growth and embryonic lethality

Micro-(mi)RNAs play a pivotal role in the developmental regulation of plants and animals. We reasoned that disruption of normal heterochronic activity in differentiating Meloidogyne incognita eggs may lead to irregular development, lethality and by extension, represent a novel target for parasite control On silencing the nuclear RNase III enzyme drosha, a critical effector of miRNA maturation in animals, we found a significant inhibition of normal development and hatching in short interfering (sORNA-soaked M incognita eggs Developing juveniles presented with highly irregular tissue patterning within the egg, and we found that unlike our previous gene silencing efforts focused on FMRFamide (Phe-Met-Arg-Phe-NH2)-like peptides (FLPs), there was no observable phenotypic recovery following removal of the environmental siRNA. Aberrant phenotypes were exacerbated over time, and drosha knockdown proved embryonically lethal Subsequently, we identified and silenced the drosha cofactor pasha, revealing a comparable inhibition of normal embryonic development within the eggs to that of drosha-silenced eggs, eventually leading to embryonic lethality To further probe the link between normal embryonic development and the M. incognita RNA interference (RNAi) pathway, we attempted to examine the impact of silencing the cytosolic RNase III enzyme dicer. Unexpectedly, we found a substantial up-regulation of dicer transcript abundance, which did not impact on egg differentiation or hatching rates. Silencing of the individual transcripts in hatched J2s was significantly less successful and resulted in temporary phenotypic aberration of the J2s. which recovered within 24 h to normal movement and posture on washing out the siRNA. Soaking the J2s in dicer siRNA resulted in a modest decrease in dicer transcript abundance which had no observable impact on phenotype or behaviour within 48 h of initial exposure to siRNA. We propose that drosha, pasha and their ancillary factors may represent excellent targets for novel nematicides and/or in planta controls aimed at M incognita, and potentially other parasitic nematodes, through disruption of miRNA-directed developmental pathways. In addition, we have identified a putative Mi-en-I transcript which encodes an RNAi-inhibiting siRNA exonuclease We observe a marked up-regulation of MI-en-I transcript abundance in response to exogenously introduced siRNA, and reason that this may impact on the interpretation of RN/NI-based reverse genetic screens in plant parasitic nematodes. (C) 2010 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Does atypical work help the jobless? Evidence from a CAEAS/CPS cohort analysis

Atypical employment, such as temporary, on-call and contract work, has been found disproportionately to attract the jobless. But there is no consensus in the literature as to the labour market consequences of such job choice by unemployed individuals. Using data from the Current Population Survey, we investigate the implications of the initial job-finding strategies pursued by the jobless for their short- and medium-term employment stability. At first sight, it appears that taking an offer of regular employment provides the greatest degree of employment continuity for the jobless. However, closer inspection indicates that the jobless who take up atypical employment are not only more likely to be employed 1 month and 1 year later than those who continue to search, but also to enjoy employment continuity that is not less favourable than that offered by regular, open-ended employment.

Crystal Creatures: Context for the Dublin Blaschka Congress

The ‘Dublin Blaschka Congress’ was conceived as a gathering to bring together the diverse scholarly disciplines that are uniquely, if eccentrically, joined in the study of scientific glass models. Leopold and Rudolf Blaschka are best known for the ‘Glass Flowers’ of Harvard but in the nineteenth century they also invented techniques to sculpt anatomically accurate marine invertebrates in glass. In the course of preparing the Congress and a coordinated temporary exhibition, much new information was uncovered about the collections of Blaschka objects in Ireland, including a total of nearly 800 surviving models. The history of the artists shows a clever business model that was designed to tap a niche market in the contemporary fascination with natural history, and improved through the course of several decades with input from clients and their own passion for understanding their biological subjects. From a modern perspective, a single Blaschka glass model of a marine invertebrate can embody biology, the history of science, craftsmanship, glass chemistry, aesthetics and art. This ability to cross interdisciplinary bridges is a singular strength of the Blaschka works, and is evident in the published proceedings of the Congress.

Welfare epidemiology as a tool to assess the welfare impact of inherited defects on the pedigree dog population

The effect that breed standards and selective breeding practices have on the welfare of pedigree dogs has recently come under scrutiny from both the general public and scientific community. Recent research has suggested that breeding for particular aesthetic traits, such as tightly curled tails, highly domed skulls and short muzzles predisposes dogs with these traits to certain inherited defects, such as spina bifida, syringomyelia and brachycephalic airway obstruction syndrome, respectively. Further to this, there is a very large number of inherited diseases that are not related to breed standards, which are thought to be prevalent, partly as a consequence of inbreeding and restricted breeding pools. Inherited diseases, whether linked to conformation or not, have varying impact on the individuals affected by them, and affect varying proportions of the pedigree dog population. Some diseases affect few breeds but are highly prevalent in predisposed breeds. Other diseases affect many breeds, but have low prevalence within each breed. In this paper, we discuss the use of risk analysis and severity diagrams as means of mapping the overall problem of inherited disorders in pedigree dogs and, more specifically, the welfare impact of specific diseases in particular breeds.

Characterization of Numb expression in astrocytomas

During early neurodevelopment, asymmetric segregation of Numb in mitotic progenitor cells influences the fate of daughter cells, whereby one daughter retains the progenitor phenotype while the other proceeds along a differentiation pathway. Numb has also been reported to function as a tumor suppressor in breast cancers and medulloblastomas. Given its role in maintaining neural progenitor pools in animal models and its reported role as a tumor suppressor, Numb could potentially contribute to astrocytoma oncogenesis. We characterized Numb expression in both human astrocytoma tissue samples and glioblastoma cell lines. We found that Numb is expressed in all grades of astrocytomas, being predominantly cytoplasmic in higher-grade astrocytomas but nuclear in pilocytic astrocytomas. Numb is also present in normal neurons, but not in normal astrocytes. In cultured glioblastoma cells, Numb concentrates in the perinuclear region and process tips. Numb expression in astrocytomas recapitulates that of progenitor cells during neurodevelopment, and suggests a role for Numb in astrocytoma oncogenesis.

Resilience and Vulnerability in the Midst of Socio-Political Violence in Northern Ireland: One Family’s Experience of a Paramilitary Style Assault.

This article focuses on the experience of one particular family living amidst the socio-political violence in Northern Ireland to illustrate the impact of a particular traumatic event – a paramilitary assault due to mistaken identity. These attacks are often colloquially referred to as a ‘punishment shootings’ or ‘beatings’. The therapeutic process is described in narrative terms, providing a framework for; understanding the systemic effect on family relationships of the initial problematic ‘storying’ of the event, and the process of ‘re-storying’ a new more coherent narrative that integrates the trauma experience. Thus, temporary family vulnerability becomes transformed into increased family resilience. This process has general applicability in work with traumatized families.

‘Towards a Green New Deal on the Island of Ireland: from economic crisis to a new political economy of sustainability’

This article begins from the assumption (which may seem controversial to many) that anyone who thinks that our current economic crisis is a temporary blip until ‘normal service’ (i.e. a return to ‘business as usual’) is resumed, profoundly misunderstands the severity and significance of what’s happening to the global economy and its impacts on the future prosperity of the island of Ireland. The economic recession represents nothing short of a re-structuring of the global economy and the creation of a new dispensation between governments, markets and citizens. The full implications of the re-regulation of the market, with the state bailing out and part nationalising the financial sector in both jurisdictions on the island (as in other parts of the world) have yet to be seen, but what we are witnessing is the emergence of a new economic model. Those who think we can, or even ought to, return to the pre-2008 economic model, are gravely mistaken. The current economic downturn marks the end of the ‘neo-liberal’ model and the beginnings of the transition (an inevitable transition, this article will argue) towards a new low carbon, renewable, green and sustainable economy and society.

Working across boundaries to improve health outcomes: a case study of a housing support and outreach service for homeless people living with HIV

This paper reports the findings of an evaluation of the ‘Housing Support, Outreach and Referral’ service developed to support people living with HIV who were homeless or at risk of homelessness. The service was set up as part of the Supporting People Health Pilot programme established to demonstrate the policy links between housing support services and health and social care services by encouraging the development of integrated services. The paper considers the role of housing support in improving people's health, and considers the challenges of working across housing, health and social care boundaries. The evaluation of the health pilot employed two main sources of data collection: quarterly project evaluation reports, which collected process data as well as reporting progress against aims and objectives, and semi-structured interviews with professionals from all key stakeholder groups and agencies, and with people who used services. Over the course of 15 months, 56 referrals were received of which 27 were accepted. Fifteen people received tenancy support of whom 12 were helped to access temporary accommodation. At the end of the 15 months, all of the tenancies had been maintained. In addition, 18 people registered with a general practitioner and 13 registered with an HIV clinic. Interviews with professionals emphasised the importance of the local joint working context, the involvement of the voluntary sector and the role of the support workers as factors that accounted for these outcomes. Those using services placed most emphasis on the flexibility of the support worker role. Importantly, interviews with professionals and those using services suggest that the role of support worker incorporates two dimensions – those of networker/navigator as well as advocate – and that both dimensions are important in determining the effectiveness of the service.

ROLE OF INTRACELLULAR AND EXTRACELLULAR CALCIUM IN HISTAMINE-RELEASE FROM RAT PERITONEAL MAST-CELLS

The present study provides evidence for a number of calcium pools important in histamine secretion from the mast cell. Firstly, calcium loosely bound to the cell membrane, and in rapid equilibrium with the extracellular environment, may be utilized for histamine release induced by most secretagogues. Secondly, all inducers are able to mobilize deeply buried or internal stores of calcium to initiate exocytosis. Finally, calcium bound to regulatory sites in the membrane may modulate the secretory process, Removal of calcium from the latter sites by brief treatment with chelating agents markedly enhances the secretory response in the absence of extracellular calcium, probably by facilitating the mobilization of bound stores of the ion, Saturation of these sites in the presence of excess calcium inhibits the release process and may restrict influx of the cation.

Critical factors affecting management of site operatives and personal in confined construction site

The aim of this paper is to identify and classify the numerous managerial issues encountered in the management of personnel in confined site construction. For the purpose of this research, a confined construction site is defined as a site where permanent works fit the site footprint, extending to levels above and/or below ground level, leaving spatial restrictions for other operations (e.g. plant and material movements, materials storage and temporary accommodation etc.) and require effective resource co-ordination beyond normal on-site management input. A literature review and analysis, case studies incorporating interviews and focus groups along with a questionnaire survey were used in order to gain a comprehensive insight into the issues in the management of personnel in a confined construction site environment. The following are the top five leading issues highlighted in the management of personnel in confined site construction; (1) Accidents due to an untidy site, (2) One contractor holding up another because of the lack of space, (3) A risk to personnel because of vehicular traffic on-site, (4) Difficult to facilitate several contractors at one work location, and (5) Numerous personnel working within the one space. In today’s modern environment, spatial restrictions are quickly becoming the norm in the industry. Therefore, the management of personnel on-site becomes progressively more difficult with the decrease in available space on-site. Where such environments exist, acknowledging the numerous issues highlighted above, aids site management in the supervision and co-ordination of personnel on-site, thus reducing accidents, increasing productivity and increase profit margins, in spatially restricted environments. As on-site management professionals successfully identify, acknowledge and counteract the numerous issues illustrated, the successful management of personnel on a confined construction site is achievable. By identifying the numerous issues, on-site management can proactively mitigate such issues through adopting counteractive measures and through successful identification of the traits identified.

Norberg-Schulz’ Elements of Space in Dark City

Alex Proyas' science fiction film Dark City (1998) puts forth questions about the building process of a city. The aliens in the film constantly alter each street, building and room to create the right environment for humans to dwell. The ‘strangers’ believe that they need to study humans in their spaces to understand human nature. They use bits and pieces of people's memories to reconstruct the city.

Christian Norberg-Schulz identifies four elements of space: physical, perceptual, existential and conceptual. Physical space is physical existence as it is. Perceptual space is the temporary space the user perceives. Existential space, for instance, the meaning of the concept of home, is abstract and permanent; it does not change with changing conditions. Finally, conceptual space, in his spatial philosophy, is the space concept of specialists like architects, economists and mathematicians.

This article analyses the future noir environment of Dark City from a spatial perspective. The notion of building is studied as to physical, perceptual, existential and conceptual spaces of Norberg-Schulz through concepts of home, identity, belonging, and alienation with reference to the architecture of the city.

Identification of Burkholderia cenocepacia genes required for bacterial survival in vivo

Burkholderia cenocepacia (formerly Burkholderia cepacia complex genomovar III) causes chronic lung infections in patients with cystic fibrosis. In this work, we used a modified signature-tagged mutagenesis (STM) strategy for the isolation of B. cenocepacia mutants that cannot survive in vivo. Thirty-seven specialized plasposons, each carrying a unique oligonucleotide tag signature, were constructed and used to examine the survival of 2,627 B. cenocepacia transposon mutants, arranged in pools of 37 unique mutants, after a 10-day lung infection in rats by using the agar bead model. The recovered mutants were screened by real-time PCR, resulting in the identification of 260 mutants which presumably did not survive within the lungs. These mutants were repooled into smaller pools, and the infections were repeated. After a second screen, we isolated 102 mutants unable to survive in the rat model. The location of the transposon in each of these mutants was mapped within the B. cenocepacia chromosomes. We identified mutations in genes involved in cellular metabolism, global regulation, DNA replication and repair, and those encoding bacterial surface structures, including transmembrane proteins and cell surface polysaccharides. Also, we found 18 genes of unknown function, which are conserved in other bacteria. A subset of 12 representative mutants that were individually examined using the rat model in competition with the wild-type strain displayed reduced survival, confirming the predictive value of our STM screen. This study provides a blueprint to investigate at the molecular level the basis for survival and persistence of B. cenocepacia within the airways.