Oral immunization with a novel lipid-based adjuvant protects against genital Chlamydia infection

Oral immunization is attractive as a delivery route because it is needle-free and useful for rapid mass vaccination programs to target pandemics or bioterrorism. This potential has not been realized for human vaccination, due to the requirement of large antigen doses and toxic (to humans) adjuvants to overcome the induction of oral tolerance and potential degradation of antigens in the stomach. To date, only oral vaccines based on live attenuated organisms have been approved for human use. In this study we describe the use of a lipid-based delivery system/adjuvant, Lipid C, for oral immunization to protect mice against genital tract chlamydial infection. Lipid C is formulated from food-grade purified and fractionated triglycerides. Bacterial shedding following vaginal challenge with Chlamydia muridarum was reduced by 50% in female mice orally immunized with the chlamydial major outer membrane protein (MOMP) formulated in Lipid C, protection equivalent to that seen in animals immunized with MOMP admixed with both cholera toxin (CT) and CpG oligodeoxynucleotides (CpG-ODN). Protection was further enhanced when MOMP, CT and CpG were all combined in the Lipid C matrix. Protection correlated with production of gamma interferon (IFN) by splenic T cells, a serum MOMP-specific IgG response and low but detectable levels of MOMP-specific IgA in vaginal lavage.

Role of Human Topoisomerase I in DNA Repair and Apoptosis

Human topoisomerase I (htopoI) is an enzyme that up to now was believed to function mainly in the removal of torsional stress generated during transcription and replication. In 1998, it was found that htopoI might play another important role in the cellular response to DNA damage -- the so-called htopoI damage response. Since this initial discovery, many studies have suggested that the htopoI damage response is involved in DNA repair as well as in apoptosis. Here we discuss the earliest as well as the latest results in this field. Combining all of the published and as yet unpublished results, we suggest and discuss a model of how htopoI may function during DNA repair and apoptosis. Furthermore, numerous results show that the htopoI damage response is not a spontaneous event, but is strictly regulated by cellular signalling pathways. We discuss which pathways may be involved and how the htopoI damage response is activated. Although the htopoI damage response was discovered several years ago, research in this area is just beginning. The future will surely bring more clarity regarding the precise mechanism behind the involvement of htopoI in DNA repair and apoptosis, as well as its implications for a broader understanding of how the human organism ensures genomic stability.

Cellular stress triggers the human topoisomerase I damage response independently of DNA damage in a p53 controlled manner

The 'human topoisomerase I (htopoI) damage response' was reported to be triggered by various kinds of DNA lesions. Also, a high and persistent level of htopoI cleavage complexes correlated with apoptosis. In the present study, we demonstrate that DNA damage-independent induction of cell death using colcemid and tumor necrosis factor is also accompanied by a strong htopoI response that correlates with the onset of apoptotic hallmarks. Consequently, these results suggest that htopoI cleavage complex formation may be caused by signaling pathways independent of the kind of cellular stress. Thus, protein interactions or signaling cascades induced by DNA damage or cellular stress might lead to the formation of stabilized cleavage complexes rather than the DNA lesion itself. Finally, we show that p53 not only plays a key role in the regulation of the htopoI response to UV-C irradiation but also to treatment with colcemid.

Genome-wide DNA methylation analysis of human brain tissue from schizophrenia patients

Recent studies suggest that genetic and environmental factors do not account for all the schizophrenia risk and epigenetics also plays a role in disease susceptibility. DNA methylation is a heritable epigenetic modification that can regulate gene expression. Genome-Wide DNA methylation analysis was performed on post-mortem human brain tissue from 24 patients with schizophrenia and 24 unaffected controls. DNA methylation was assessed at over 485 000 CpG sites using the Illumina Infinium Human Methylation450 Bead Chip. After adjusting for age and post-mortem interval (PMI), 4 641 probes corresponding to 2 929 unique genes were found to be differentially methylated. Of those genes, 1 291 were located in a CpG island and 817 were in a promoter region. These include NOS1, AKT1, DTNBP1, DNMT1, PPP3CC and SOX10 which have previously been associated with schizophrenia. More than 100 of these genes overlap with a previous DNA methylation study of peripheral blood from schizophrenia patients in which 27 000 CpG sites were analysed. Unsupervised clustering analysis of the top 3 000 most variable probes revealed two distinct groups with significantly more people with schizophrenia in cluster one compared to controls (p = 1.74x10-4). The first cluster was composed of 88% of patients with schizophrenia and only 12% controls while the second cluster was composed of 27% of patients with schizophrenia and 73% controls. These results strongly suggest that differential DNA methylation is important in schizophrenia etiology and add support for the use of DNA methylation profiles as a future prognostic indicator of schizophrenia.

A novel method using intranasal delivery of EdU demonstrates that accessory olfactory ensheathing cells respond to injury by proliferation

Olfactory ensheathing cells (OECs) play an important role in the continuous regeneration of the primary olfactory nervous system throughout life and for regeneration of olfactory neurons after injury. While it is known that several individual OEC subpopulations with distinct properties exist in different anatomical locations, it remains unclear how these different subpopulations respond to a major injury. We have examined the proliferation of OECs from one distinct location, the peripheral accessory olfactory nervous system, following large-scale injury (bulbectomy) in mice. We used crosses of two transgenic reporter mouse lines, S100ß-DsRed and OMP-ZsGreen, to visualise OECs, and main/accessory olfactory neurons, respectively. We surgically removed one olfactory bulb including the accessory olfactory bulb to induce degeneration, and found that accessory OECs in the nerve bundles that terminate in the accessory olfactory bulb responded by increased proliferation with a peak occurring 2 days after the injury. To label proliferating cells we used the thymidine analogue ethynyl deoxyuridine (EdU) using intranasal delivery instead of intraperitoneal injection. We compared and quantified the number of proliferating cells at different regions at one and four days after EdU labelling by the two different methods and found that intranasal delivery method was as effective as intrapeitoneal injection. We demonstrated that accessory OECs actively respond to widespread degeneration of accessory olfactory axons by proliferating. These results have important implications for selecting the source of OECs for neural regeneration therapies and show that intranasal delivery of EdU is an efficient and reliable method for assessing proliferation of olfactory glia.

The mouse model of Chlamydia genital tract infection: a review of infection, disease, immunity and vaccine development

Chlamydia trachomatis is the most common sexually transmitted bacterial infection worldwide. The impact of this pathogen on human reproduction has intensified research efforts to better understand chlamydial infection and pathogenesis. Whilst there are animal models available that mimic the many aspects of human chlamydial infection, the mouse is regarded as the most practical and widely used of the models. Studies in mice have greatly contributed to our understanding of the host-pathogen interaction and provided an excellent medium for evaluating vaccines. Here we explore the advantages and disadvantages of all animal models of chlamydial genital tract infection, with a focus on the murine model and what we have learnt from it so far.

Immunization with a MOMP-based vaccine protects mice against a Pulmonary Chlamydia challenge and identifies a disconnection between infection and pathology

Chlamydia pneumoniae is responsible for up to 20% of community acquired pneumonia and can exacerbate chronic inflammatory diseases. As the majority of infections are either mild or asymptomatic, a vaccine is recognized to have the greatest potential to reduce infection and disease prevalence. Using the C. muridarum mouse model of infection, we immunized animals via the intranasal (IN), sublingual (SL) or transcutaneous (TC) routes, with recombinant chlamydial major outer membrane protein (MOMP) combined with adjuvants CTA1-DD or a combination of cholera toxin/CpG-oligodeoxynucleotide (CT/CpG). Vaccinated animals were challenged IN with C. muridarum and protection against infection and pathology was assessed. SL and TC immunization with MOMP and CT/CpG was the most protective, significantly reducing chlamydial burden in the lungs and preventing weight loss, which was similar to the protection induced by a previous live infection. Unlike a previous infection however, these vaccinations also provided almost complete protection against fibrotic scarring in the lungs. Protection against infection was associated with antigen-specific production of IFNγ, TNFα and IL-17 by splenocytes, however, protection against both infection and pathology required the induction of a similar pro-inflammatory response in the respiratory tract draining lymph nodes. Interestingly, we also identified two contrasting vaccinations capable of preventing infection or pathology individually. Animals IN immunized with MOMP and either adjuvant were protected from infection, but not the pathology. Conversely, animals TC immunized with MOMP and CTA1-DD were protected from pathology, even though the chlamydial burden in this group was equivalent to the unimmunized controls. This suggests that the development of pathology following an IN infection of vaccinated animals was independent of bacterial load and may have been driven instead by the adaptive immune response generated following immunization. This identifies a disconnection between the control of infection and the development of pathology, which may influence the design of future vaccines.

Primary schools and the delivery of relationships and sexuality education : the experience of Queensland teachers

Primary school provides an appropriate opportunity for children to commence comprehensive relationships and sexuality education (RSE), yet many primary school teachers avoid teaching this subject area. In the absence of teacher confidence and competence, schools have often relied on health promotion professionals, external agencies and/or one-off issue related presentations rather than cohesive, systematic and meaningful health education. This study examines the implementation of a ten-lesson pilot RSE unit of work and accompanying assessment task in two primary schools in South-East Queensland, Australia. Drawing predominantly from qualitative data, this research explores the experiences of primary school teachers as they engage with RSE curriculum resources and content delivery. The results show that the provision of a high quality RSE curriculum resource grounded in contemporary educational principles and practices enables teachers to feel more confident to deliver RSE and minimises potential barriers such as parental objections and fear of mishandling sensitive content.

The salt dependence of DNA recognition by NF-kappaB p50 : a detailed kinetic analysis of the effects on affinityand specificity

The binding kinetics of NF-kappaB p50 to the Ig-kappaB site and to a DNA duplex with no specific binding site were determined under varying conditions of potassium chloride concentration using a surface plasmonresonance biosensor. Association and dissociation rate constants were measured enabling calculation of the dissociation constants. Under previously established high affinity buffer conditions, the k a for both sequences was in the order of 10(7) M-1s-1whilst the k d values varied 600-fold in a sequence-dependent manner between 10(-1) and 10(-4 )s-1, suggesting that the selectivity of p50 for different sequences is mediated primarily through sequence-dependent dissociation rates. The calculated K D value for the Ig-kappaB sequence was 16 pM, whilst the K D for the non-specific sequence was 9.9 nM. As the ionic strength increased to levels which are closer to that of the cellular environment, the binding of p50 to the non-specific sequence was abolished whilst the specific affinity dropped to nanomolar levels. From these results, a mechanism is proposed in which p50 binds specific sequences with high affinity whilst binding non-specific sequences weakly enough to allow efficient searching of the DNA.

Distamycin A affects the stability of NF-kappaB p50-DNA complexes in a sequence-dependent manner

The effect of two different DNA minor groove binding molecules, Hoechst 33258 and distamycin A, on the binding kinetics of NF-κB p50 to three different specific DNA sequences was studied at various salt concentrations. Distamycin A was shown to significantly increase the dissociation rate constant of p50 from the sequences PRDII (5′-GGGAAATTCC-3′) and Ig-κ B (5′-GGGACTTTCC-3′) but had a negligible effect on the dissociation from the palindromic target-κB binding site (5′-GGGAATTCCC-3′). By comparison, the effect of Hoechst 33258 on binding of p50 to each sequence was found to be minimal. The dissociation rates for the protein–DNA complexes increased at higher potassium chloride concentrations for the PRDII and Ig-κB binding motifs and this effect was magnified by distamycin A. In contrast, p50 bound to the palindromic target-κB site with a much higher intrinsic affinity and exhibited a significantly reduced salt dependence of binding over the ionic strength range studied, retaining a KD of less than 10 pM at 150 mM KCl. Our results demonstrate that the DNA binding kinetics of p50 and their salt dependence is strongly sequence-dependent and, in addition, that the binding of p50 to DNA can be influenced by the addition of minor groove-binding drugs in a sequence-dependent manner.

Research roadmap summary - Integrated Design and Delivery Solutions (IDDS)

Changes in the construction sector are creating opportunities in research to maximise the benefits of those changes and to continue the exciting developments in improved people skills, new processes and developing technologies. Many research centres around the world are investigating aspects of the current changes to drive their particular expertise forward. However, the CIB Integrated Design and Delivery Solutions (IDDS) priority research theme takes a higher-level view of the changes and then focuses down on a prioritised set of research targets. These targets have been investigated, re-focussed and validated over a period of four years through many workshops, conferences and meetings by a wide ranging group of representatives from approximately 90 industry and research organisations. The outcomes of such research, once put into practice should be significantly shortened timespans from conception of need to occupation of new or revised structures. As time is money, the owners will get their investments into productive use sooner, which means a shorter payback time. In addition, there will inevitably be a reduction in construction costs as productivity increases. The improvements in reliable delivery and improved quality currently being seen in relatively simplistic use of Building information Modelling (BIM) (compared to full IDDS) will inevitably continue its on-going trajectory of improvement. We should also consider the wider economic contribution to society that will stem from such improvements and, finally, and by no means unimportantly, the reliable modelling and delivery of sustainability at both the building and estate/ area scale will significantly improve carbon footprints and other sustainable outcomes. Whilst there are huge opportunities for early adopters, the primary risk will be the expansion of the gap between those working in this way and those who are not so advanced or who even refuse to progress . The opportunities to address the significant and widely varying wastes within the structure of the construction sector and within and across projects are huge and timely and industry is encouraged to become involved.

Integrated design and delivery solutions

Integrated design and delivery solutions (IDDS) is a priority theme of the International Council for Research and Innovation in Building and Construction (CIB), which will be used to drive the global research agenda forward. IDDS will use collaborative work processes and enhanced skills together with integrated data, information and knowledge management to minimize structural and process inefficiencies and to enhance the value delivered during design, build, operation, and across projects. IDDS build on building information modelling (BIM), incorporating advances in the training and employment of people, together with supporting new technologies. The successful use of IDDS involves changes in each of the project phases from conceptual planning and business case formulation to all stages of the supply chain: design, construction, commissioning, operation, retrofit and decommissioning. For each of these phases, key changes in the structure and culture of the project team across the different collaborating firms create a favourable context for IDDS. Special for IDDS thinking is the idea of adding project and whole-life value in all phases, for all stakeholders...

Challenges for integrated design and delivery solutions

A new approach of integrated design and delivery solutions (IDDS) aims to radically improve the performance of the construction industries. IDDS builds upon recent trends in the construction industries that have seen the widespread adoption of technologies such as building information modelling (BIM) and innovative processes such as integrated project delivery. However, these innovations are seen to develop in isolation, with little consideration of the overarching interactions between people, process and technology. The IDDS approach is holistic in that it recognizes that it is only through a combination of initiatives such as skill development, process re-engineering, responsive information technology, enhanced interoperability and integrating knowledge management, among others, that radical change can be achieved. To implement IDDS requires step changes in many project aspects, and this gap between current performance and that required for IDDS is highlighted. The research required to bridge the gaps is identified in four major aspects of collaborative processes, workforce skills, integrated information and knowledge management.

CIB White Paper on IDDS Integrated Design & Delivery Solutions

CIB is developing a priority theme, now termed Improving Construction and Use through Integrated Design & Delivery Solutions (IDDS). The IDDS working group for this theme adopted the following definition: Integrated Design and Delivery Solutions use collaborative work processes and enhanced skills, with integrated data, information, and knowledge management to minimize structural and process inefficiencies and to enhance the value delivered during design, build, and operation, and across projects. The design, construction, and commissioning sectors have been repeatedly analysed as inefficient and may or may not be quite as bad as portrayed; however, there is unquestionably significant scope for IDDS to improve the delivery of value to clients, stakeholders (including occupants), and society in general, simultaneously driving down cost and time to deliver operational constructed facilities. Although various initiatives developed from computer‐aided design and manufacturing technologies, lean construction, modularization, prefabrication and integrated project delivery are currently being adopted by some sectors and specialisations in construction; IDDS provides the vision for a more holistic future transformation. Successful use of IDDS requires improvements in work processes, technology, and people’s capabilities to span the entire construction lifecycle from conception through design, construction, commissioning, operation, refurbishment/ retrofit and recycling, and considering the building’s interaction with its environment. This vision extends beyond new buildings to encompass modifications and upgrades, particularly those aimed at improved local and area sustainability goals. IDDS will facilitate greater flexibility of design options, work packaging strategies and collaboration with suppliers and trades, which will be essential to meet evolving sustainability targets. As knowledge capture and reuse become prevalent, IDDS best practice should become the norm, rather than the exception.

Research roadmap report : Integrated Design and Delivery Solutions (IDDS)

Changes in the construction sector are creating opportunities in research to maximise the benefits of those changes and to continue the exciting developments in improved people skills, new processes and developing technologies. There are many research centres around the world investigating aspects of the current changes to drive their particular expertise forward. However, the CIB Integrated Design and Delivery Solutions (IDDS) priority research theme takes a higher-level view of the changes and then focuses down on a prioritised set of research targets. These targets have been investigated, re-focussed and validated over a period of four years through many workshops, conferences and meetings by a wide ranging group of representatives from approximately 90 industry and research organisations. This roadmap prioritises and details the research to be performed, why and by whom. In particular, some 25 CIB Working Commissions and Task Groups are explained as having potential roles in the delivery of this research theme. We are extremely privileged to have been urged on by such distinguished construction professionals in their forewords and the case for research. The outcomes of such research, once put into practice should be significantly shortened timespans from conception of need to occupation of new or revised structures. As time is money, the owners will get their investments into productive use sooner, which means a shorter payback time. In addition, there will inevitably be a reduction in construction costs as productivity increases. The improvements in reliable delivery and improved quality currently being seen in relatively simplistic use of Building information Modelling (BIM) (compared to full IDDS) will inevitably continue its on-going trajectory of improvement. We should also consider the wider economic contribution to society that will stem from such improvements and, finally, and by no means unimportantly, the reliable modelling and delivery of sustainability at both the building and estate/ area scale will significantly improve carbon footprints and other sustainable outcomes. Whilst there are huge opportunities for early adopters, the primary risk will be the expansion of the gap between those working in this way and those who are not so advanced or who even refuse to progress1. However, a similar issue arises between industry, clients, educators and trainers; the latter have particular challenges, having existed for many years in a sector that has had relatively few technological changes. However, the opportunities to address the significant and widely varying wastes within the structure of the construction sector and within and across projects are huge and timely. Whilst this Roadmap is specifically targeted at the Standing Commissions and Task Groups of the CIB, it is hoped that there are elements for research and applied research across academia and industry.