Innate transcriptional networks activated in bladder in response to uropathogenic Escherichia coli drive diverse biological pathways and rapid synthesis of IL-10 for defense against bacterial urinary tract infection

Early transcriptional activation events that occur in bladder immediately following bacterial urinary tract infection (UTI) are not well defined. In this study, we describe the whole bladder transcriptome of uropathogenic Escherichia coli (UPEC) cystitis in mice using genome-wide expression profiling to define the transcriptome of innate immune activation stemming from UPEC colonization of the bladder. Bladder RNA from female C57BL/6 mice, analyzed using 1.0 ST-Affymetrix microarrays, revealed extensive activation of diverse sets of innate immune response genes, including those that encode multiple IL-family members, receptors, metabolic regulators, MAPK activators, and lymphocyte signaling molecules. These were among 1564 genes differentially regulated at 2 h postinfection, highlighting a rapid and broad innate immune response to bladder colonization. Integrative systems-level analyses using InnateDB (http://www.innatedb.com) bioinformatics and ingenuity pathway analysis identified multiple distinct biological pathways in the bladder transcriptome with extensive involvement of lymphocyte signaling, cell cycle alterations, cytoskeletal, and metabolic changes. A key regulator of IL activity identified in the transcriptome was IL-10, which was analyzed functionally to reveal marked exacerbation of cystitis in IL-10–deficient mice. Studies of clinical UTI revealed significantly elevated urinary IL-10 in patients with UPEC cystitis, indicating a role for IL-10 in the innate response to human UTI. The whole bladder transcriptome presented in this work provides new insight into the diversity of innate factors that determine UTI on a genome-wide scale and will be valuable for further data mining. Identification of protective roles for other elements in the transcriptome will provide critical new insight into the complex cascade of events that underpin UTI.

Project-based civil engineering courses

Problem solving is an essential element of civil engineering education. It has been observed that students are best able to understand civil engineering theory when there is a practical application of it. Teaching theory alone has led to lower levels of comprehension and motivation and a correspondingly higher rate of failure and “drop-out”. This paper analyses the effectiveness of introducing practical design projects at an early stage within a civil engineering undergraduate program at Queensland University of Technology. In two of the essential basic subjects, Engineering Mechanics and Steel Structures, model projects which simulate realistic engineering exercises were introduced. Students were required to work in small groups to analyse, design and build the lightest / most efficient model bridges made of specific materials such as spaghetti, drinking straw, paddle pop sticks and balsa wood and steel columns for a given design loading/target capacity. The paper traces the success of the teaching strategy at each stage from its introduction through to the final student and staff evaluation.

Governing civil society: How literacy, education and security were brought together

This study questions how the categories of security, education and literacy were brought together as related elements of a whole-of-government strategy in the production of civil society. Drawing on an analysis of key political texts, the study argues that the categories of education and literacy have been used in diverse ways in the production of national, social, economic and geopolitical security interests. As dialogue about security has intensified, rationalisations about the national interest have engaged notions of security leading to the legitimation of a diverse set of policy instruments, strategically used to contain the rise of complex social forces and protect homogenous cultural values.

Time and cost performance of design–build projects

The design-build (DB) delivery method has been widely used in the United States due to its reputed superior cost and time performance. However, rigorous studies have produced inconclusive support and only in terms of overall results, with few attempts being made to relate project characteristics with performance levels. This paper provides a larger and more finely grained analysis of a set of 418 DB projects from the online project database of the Design-Build Institute of America (DBIA), in terms of the time-overrun rate (TOR), early start rate (ESR), early completion rate (ECR) and cost overrun rate (COR) associated with project type (e.g., commercial/institutional buildings and civil infrastructure projects), owners (e.g., Department of Defense and private corporations), procurement methods (e.g., ‘best value with discussion’ and qualifications-based selection), contract methods (e.g., lump sum and GMP) and LEED levels (e.g., gold and silver). The results show ‘best value with discussion’ to be the dominant procurement method and lump sum the most frequently used contract method. The DB method provides relatively good time performance, with more than 75% of DB projects completed on time or before schedule. However, with more than 50% of DB projects cost overrunning, the DB advantage of cost saving remains uncertain. ANOVA tests indicate that DB projects within different procurement methods have significantly different time performance and that different owner types and contract methods significantly affect cost performance. In addition to contributing to empirical knowledge concerning the cost and time performance of DB projects with new solid evidence from a large sample size, the findings and practical implications of this study are beneficial to owners in understanding the likely schedule and budget implications involved for their particular project characteristics.

Infection and cellular defense dynamics in a novel 17beta-estradiol murine model of chronic human group B streptococcus genital tract colonization reveal a role for hemolysin in persistence and neutrophil accumulation

Genital tract carriage of group B streptococcus (GBS) is prevalent among adult women; however, the dynamics of chronic GBS genital tract carriage, including how GBS persists in this immunologically active host niche long term, are not well defined. To our knowledge, in this study, we report the first animal model of chronic GBS genital tract colonization using female mice synchronized into estrus by delivery of 17β-estradiol prior to intravaginal challenge with wild-type GBS 874391. Cervicovaginal swabs, which were used to measure bacterial persistence, showed that GBS colonized the vaginal mucosa of mice at high numbers (106–107 CFU/swab) for at least 90 d. Cellular and histological analyses showed that chronic GBS colonization of the murine genital tract caused significant lymphocyte and PMN cell infiltrates, which were localized to the vaginal mucosal surface. Long-term colonization was independent of regular hormone cycling. Immunological analyses of 23 soluble proteins related to chemotaxis and inflammation showed that the host response to GBS in the genital tract comprised markers of innate immune activation including cytokines such as GM-CSF and TNF-α. A nonhemolytic isogenic mutant of GBS 874391, Δcyle9, was impaired for colonization and was associated with amplified local PMN responses. Induction of DNA neutrophil extracellular traps, which was observed in GBS-infected human PMNs in vitro in a hemolysin-dependent manner, appeared to be part of this response. Overall, this study defines key infection dynamics in a novel murine model of chronic GBS genital tract colonization and establishes previously unknown cellular and soluble defense responses to GBS in the female genital tract.

Vaccination of koalas (Phascolarctos cinereus) with a recombinant chlamydial major outer membrane protein adjuvanted with poly I:C, a host defense peptide and polyphosphazine, elicits strong and long lasting cellular and humoral immune responses

Chlamydial infections are wide spread in koalas across their range and a solution to this debilitating disease has been sought for over a decade. Antibiotics are the currently accepted therapeutic measure, but are not an effective treatment due to the asymptomatic nature of some infections and a low efficacy rate. Thus, a vaccine would be an ideal way to address this infectious disease threat in the wild. Previous vaccine trials have used a three-dose regimen; however this is very difficult to apply in the field as it would require multiple capture events, which are stressful and invasive processes for the koala. In addition, it requires skilled koala handlers and a significant monetary investment. To overcome these challenges, in this study we utilized a polyphosphazine based poly I:C and a host defense peptide adjuvant combined with recombinant chlamydial major outer membrane protein (rMOMP) antigen to induce long lasting (54 weeks) cellular and humoral immunity in female koalas with a novel single immunizing dose. Immunized koalas produced a strong IgG response in plasma, as well as at mucosal sites. Moreover, they showed high levels of C. pecorum specific neutralizing antibodies in the plasma as well as vaginal and conjunctival secretions. Lastly, Chlamydia-specific lymphocyte proliferation responses were produced against both whole chlamydial elementary bodies and rMOMP protein, over the 12-month period. The results of this study suggest that a single dose rMOMP vaccine incorporating a poly I:C, host defense peptide and polyphosphazine adjuvant is able to stimulate both arms of the immune system in koalas, thereby providing an alternative to antibiotic treatment and/or a three-dose vaccine regime.

Medical negligence, causation and 'exceptional cases' under the civil liability legislation

Under the civil liability legislation enacted in most Australian jurisdictions, factual causation will be established if, on the balance of probabilities, the claimant can prove that the defendant's negligence was 'a necessary condition of the occurrence of the [claimant's] harm'. Causation will then be satisfied by showing that the harm would not have occurred 'but for' the defendant's breach of their duty of care. However, in an exceptional or appropriate case, sub-section 2 of the legislation provides that if the 'but for' test is not met, factual causation may instead be determined in accordance with other 'established principles'. In such a case, 'the court is to consider (amongst other relevant things) whether or not and why responsibility for the harm should be imposed' on the negligent party.

Civil service HRM reforms in a new democracy: The case of Bhutan

This research investigates Bhutan Civil Service Human Resource Management strategies, policies and practices, and their contribution to achieving the national goal of Gross National Happiness. The study finds that the HRM of the Bhutanese civil service is meeting its strategic objective of contributing to GNH. The civil service in Bhutan plays an important role in socio-economic development, influences private sector practices, strengthens good governance and provides continuity to the government. Participants in the study were government ministers and senior, highly experienced civil servants. A model of civil service HRM in Bhutan is developed.

Submission on NSW Attorney-General's Discussion Paper on Amendment of Limitation of Actions Act 1969 to remove time limit for civil claims for child abuse

The New South Wales Attorney-General and Justice Policy Division released a Discussion Paper about reform of the Limitation of Actions Act 1969. The key question was whether and how to amend the statute to better provide access to justice for civil claimants in child abuse cases. This submission draws on published literature and multidisciplinary research to support the Discussion Paper's Option A, namely, to abolish the time limit for civil claims for injuries in criminal child abuse cases, and for this to be made retrospective.

Civil Procedure — Commentary and Materials, 6th Edition

This book provides a comprehensive analysis of the practical and theoretical issues encountered in Australian civil procedure, including alternative dispute resolution. Each chapter features in-depth questions and notes together with lists of further reading to aid understanding of the issue. It also examines and discusses each substantive and procedural step in the trial process. Topics include jurisdiction of a court to consider a matter, alternative dispute resolution, limitations of actions, commencing proceedings, group proceedings, pleading, summary disposition, gathering evidence, affidavits, interlocutory procedures, settlement, trial and appeal, costs Each of the state, territory and federal procedures is covered.

Models for engaging public-private partnerships in civil infrastructure projects: A case of ‘having your cake and eating it too’?

Public-Private Partnerships (PPP) are established globally as an important mode of procurement and the features of PPP, not least of which the transfer of risk, appeal to governments and particularly in the current economic climate. There are many other advantages of PPP that are claimed as outweighing the costs of PPP and affording Value for Money (VfM) relative to traditionally financed projects or non-PPP. That said, it is the case that we lack comparative whole-life empirical studies of VfM in PPP and non-PPP. Whilst we await this kind of study, the pace and trajectory of PPP seem set to continue and so in the meantime, the virtues of seeking to improve PPP appear incontrovertible. The decision about which projects, or parts of projects, to offer to the market as a PPP and the decision concerning the allocation or sharing risks as part of engagement of the PPP consortium are among the most fundamental decisions that determine whether PPP deliver VfM. The focus in the paper is on latter decision concerning governments’ attitudes towards risk and more specifically, the effect of this decision on the nature of the emergent PPP consortium, or PPP model, including its economic behavior and outcomes. This paper presents an exploration into the extent to which the seemingly incompatible alternatives of risk allocation and risk sharing, represented by the orthodox/conventional PPP model and the heterodox/alliance PPP model respectively, can be reconciled along with suggestions for new research directions to inform this reconciliation. In so doing, an important step is taken towards charting a path by which governments can harness the relative strengths of both kinds of PPP model.

Genome-wide mapping of cystitis due to Streptococcus agalactiae and Escherichia coli in mice identifies a unique bladder transcriptome that signifies pathogen-specific antimicrobial defense against urinary tract infection

The most common causes of urinary tract infections (UTIs) are Gram-negative pathogens such as Escherichia coli; however, Gram-positive organisms including Streptococcus agalactiae, or group B streptococcus (GBS), also cause UTI. In GBS infection, UTI progresses to cystitis once the bacteria colonize bladder, but the host responses triggered in the bladder immediately following infection are largely unknown. Here, we used genome-wide expression profiling to map the bladder transcriptome of GBS UTI in mice infected transurethrally with uropathogenic GBS that was cultured from a 35 year-old women with cystitis. RNA from bladders was applied to Affymetrix Gene-1.0ST microarrays; qRT-PCR was used to analyze selected gene responses identified in array datasets. A surprisingly small significant gene list of 172 genes was identified at 24h; this compared to 2507 genes identified in a side-by-side comparison with uropathogenic E. coli (UPEC). No genes exhibited significantly altered expression at 2h in GBS-infected mice according to arrays despite high bladder bacterial loads at this early time point. The absence of a marked early host response to GBS juxtaposed with broad-based bladder responses activated by UPEC at 2h. Bioinformatics analyses including integrative systems-level network mapping revealed multiple activated biological pathways in the GBS cystitis transcriptome that regulate leukocyte activation, inflammation, apoptosis, and cytokine-chemokine biosynthesis. These findings define a novel, minimalistic type of bladder host response triggered by GBS UTI, which comprises collective antimicrobial pathways that differ dramatically from those activated by UPEC. Overall, this study emphasizes the unique nature of bladder immune activation mechanisms triggered by distinct uropathogens.