Validation of an immunoassay to measure plasminogen-activator inhibitor-1 concentrations in human saliva

Introduction We have previously shown that the concentrations of D-dimer are significantly elevated in saliva compared with plasma. Saliva offers several advantages compared with blood analysis. We hypothesised that human saliva contains plasminogen activator inhibitor-1 (PAI-1) and that the concentrations are not affected by the time of saliva collection. The aim was to adopt and validate an immunoassay to quantify PAI-1 concentrations in saliva and to determine whether saliva collection time has an influence in the measurement. Materials and methods Two saliva samples (morning and afternoon) from the same day were collected from healthy subjects (N = 40) who have had no underlying heart conditions. A customized AlphaLISA® immunoassay (PerkinElmer®, MA, USA) was adopted and used to quantify PAI-1 concentrations. We validated the analytical performance of the customized immunoassay by calculating recovery of known amount of analyte spiked in saliva. Results: The recovery (95.03%), intra- (8.59%) and inter-assay (7.52%) variations were within the acceptable ranges. The median salivary PAI-1 concentrations were 394 pg/mL (interquartile ranges (IQR) 243.4-833.1 pg/mL) in the morning and 376 (129.1-615.4) pg/mL in the afternoon and the plasma concentration was 59,000 (24,000-110,000) pg/mL. Salivary PAI-1 did not correlate with plasma (P = 0.812). Conclusions The adopted immunoassay produced acceptable assay sensitivity and specificity. The data demonstrated that saliva contains PAI-1 and that its concentration is not affected by the time of saliva collection. There is no correlation between salivary and plasma PAI-1 concentrations. Further studies are required to demonstrate the utility of salivary PAI-1 in CVD risk factor studies.

Obesity and hypertension have differing oxidant handling molecular pathways in age-related chronic kidney disease

Chronic kidney disease (CKD) in ageing is a burden on health systems worldwide. Rat models of age-related CKD linked with obesity and hypertension were used to investigate alterations in oxidant handling and energy metabolism to identify gene targets or markers for age-related CKD. Young adult (3 months) and old (21–24 months) spontaneously-hypertensive (SHR), normotensive Wistar-Kyoto (WKY) and Wistar rats (normotensive, obese in ageing) were compared for renal functional and physiological parameters, renal fibrosis and inflammation, oxidative stress (hemeoxygenase-1/HO-1), apoptosis and cell injury (including Bax:Bcl-2), phosphorylated and non-phosphorylated forms of oxidant and energy sensing proteins (p66Shc, AMPK), signal transduction proteins (ERK1/2, PKB), and transcription factors (NF-κB, FoxO1). All old rats were normoglycemic. Renal fibrosis, tubular epithelial apoptosis, interstitial macrophages and myofibroblasts (all p < 0.05), p66Shc/phospho-p66 (p < 0.05), Bax/Bcl-2 ratio (p < 0.05) and NF-κB expression (p < 0.01) were highest in old obese Wistars. Expression of phospho-FoxO/FoxO was elevated in old Wistars (p < 0.001) and WKYs (p < 0.01). SHRs had high levels in young and old rats. Expression of PKB, phospho-PKB, ERK1/2 and phospho-ERK1/2 were significantly elevated in all aged animals. These results suggest that obesity and hypertension have differing oxidant handling and signalling pathways that act in the pathogenesis of age-related CKD

Into darkness: A study of deviance in Star Trek

Purpose The purpose of this chapter is to analyse the deviant behaviour of individuals in organisations. Deviants are those who depart from organisational norms. A typology of perceived deviant behaviour is developed from the deviance literature, and subsequently tested. Methodology/approach Star Trek: Into Darkness text is qualitatively analysed as a data source. Three different character arcs are analysed in relation to organisational deviance. Starfleet is the specific, fictional, organisational context. Findings We found that the typology of deviance is conceptually robust, and facilitates categorisation of different types of deviant behaviour, over time. Research limitations/implications Deviance is socially ascribed; so better categorisation of such behaviour improves our understanding of how specific behaviour might deviate from organisational norms, and how different behaviours can mean individuals can be viewed positively or negatively over time. Further research might determine management responses to the different forms of deviance, and unpack the processes where individuals eschew ‘averageness’ and become deviants. Practical implications The typology advanced has descriptive validity to describe deviant behaviour. Social implications Social institutions such as organisations ascribe individual deviants, both negatively and positively. Originality/value This chapter extends our understanding of positive and negative deviance in organisations by developing a new typology of deviant behaviour. This typology has descriptive validity in understanding deviant behaviour. Our understanding of both positive and negative deviance in organisational contexts is enhanced, as well as the utility of science fiction literature in ethical analysis.

Regulation, compliance and the Australian forensic accounting profession

Legacies of the Global Financial Crisis and major domestic corporate collapses – such as HIH Insurance Pty Ltd and One.Tel Ltd (telecommunications) – have significantly changed Australia‟s financial regulatory landscape. Legal requirements for auditors have attracted particular attention as have practice standards more broadly around disclosure and conflict of interest. Conversely, although successful detection and prosecution of breaches may rest in significant part on forensic accounting activities, Australia‟s practitioners in this field have no minimum training or qualifications standards other than the baseline requirements mandated by the country‟s three professional accounting bodies. For those unaffiliated with these organizations, no professional oversight exists. In Australia, growth in the forensic accounting industry has been in direct response to public demand for expertise in a broad range of fraud, forensic and business analytics areas in order to improve the corporate governance practices of Australian organizations. During the 1990s, Australian forensic accounting firms expanded and diversified into a number of different areas going well beyond just the examination of financial documents and involvement in financial litigation disputes. “Big 4” accounting firms such as PriceWaterhouseCoopers, KPMG, Deloitte and Ernst and Young formed independent forensic accounting or forensic services units; a number of mid-tier and „boutique‟ forensic accounting firms similarly expanded into forensic investigative, analytical and advisory services. By 2008, 800 forensic accountants were registered with the country‟s largest specialist forensic accounting group, the Forensic Accounting Special Interest Group (FASIG) of the ICAA1. Currently, obtaining more precise figures on numbers of forensic accounting practitioners is problematic: professional accounting bodies either do not keep a register or have ceased registering their forensic accounting members; lack of formal recognition, admission or certification processes complicate identification of candidates; and diversity of the skills sets the industry requires has meant the influx of non-accounting based specialists.

Significance of drying periods on nitrate removal in experimental biofilters

Nitrogen is an important nutrient that can impact the quality of aquatic environments when present in high concentration. Even though low concentration levels of ammonium-nitrogen have been observed in laboratory studies in bioretention basins, poor removal or even the production of nitrate-nitrogen within the filter is often recorded in such studies. Ten Perspex biofilter columns of 94 mm (internal diameter) were packed with a filter layer, transition layer and a gravel layer. While the filter layer was packed to a height of 800 mm, transition and gravel layers were packed to a composite height of 220 mm and operated with simulated stormwater in the laboratory. The filter layer contained 8% organic material by weight. A free board of 350 mm provided detention storage and head to facilitate infiltration. The columns were operated with different antecedent dry days (0 d to 21 d) and constant inflow concentration at a feed rate of 100 mL/min. Samples were collected from the outflow at different time intervals, between 2 min and 150 min from the start of outflow, and were tested for nitrate-nitrogen and total organic carbon. Washoff of organic carbon from the filter layer was observed to occur for 30 min of outflow. This indicated washoff of organic carbon from the filter itself. At the same time, a very low concentration of nitrate-nitrogen was recorded at the beginning of the outflow, indicating the effective removal of nitrate-nitrogen. We conclude that the removal of nitrate-nitrogen is insignificant during the wetting phase of a rainfall event and the process of denitrification is more pronounced during the drying phase of a rainfall event. Thus intermittent wetting and drying is crucial for the removal of nitrate-nitrogen in bioretention basins.

Deriving a preference-based utility measure for cancer patients from the European Organisation for the Research and Treatment of Cancer's Quality of Life Questionnaire C30 : a confirmatory versus exploratory approach

Background Multi attribute utility instruments (MAUIs) are preference-based measures that comprise a health state classification system (HSCS) and a scoring algorithm that assigns a utility value to each health state in the HSCS. When developing a MAUI from a health-related quality of life (HRQOL) questionnaire, first a HSCS must be derived. This typically involves selecting a subset of domains and items because HRQOL questionnaires typically have too many items to be amendable to the valuation task required to develop the scoring algorithm for a MAUI. Currently, exploratory factor analysis (EFA) followed by Rasch analysis is recommended for deriving a MAUI from a HRQOL measure. Aim To determine whether confirmatory factor analysis (CFA) is more appropriate and efficient than EFA to derive a HSCS from the European Organisation for the Research and Treatment of Cancer’s core HRQOL questionnaire, Quality of Life Questionnaire (QLQ-C30), given its well-established domain structure. Methods QLQ-C30 (Version 3) data were collected from 356 patients receiving palliative radiotherapy for recurrent/metastatic cancer (various primary sites). The dimensional structure of the QLQ-C30 was tested with EFA and CFA, the latter informed by the established QLQ-C30 structure and views of both patients and clinicians on which are the most relevant items. Dimensions determined by EFA or CFA were then subjected to Rasch analysis. Results CFA results generally supported the proposed QLQ-C30 structure (comparative fit index =0.99, Tucker–Lewis index =0.99, root mean square error of approximation =0.04). EFA revealed fewer factors and some items cross-loaded on multiple factors. Further assessment of dimensionality with Rasch analysis allowed better alignment of the EFA dimensions with those detected by CFA. Conclusion CFA was more appropriate and efficient than EFA in producing clinically interpretable results for the HSCS for a proposed new cancer-specific MAUI. Our findings suggest that CFA should be recommended generally when deriving a preference-based measure from a HRQOL measure that has an established domain structure.

Double-strand breaks and the concept of short- and long-term epigenetic memory

Double-strand breaks represent an extremely cytolethal form of DNA damage and thus pose a serious threat to the preservation of genetic and epigenetic information. Though it is well-known that double-strand breaks such as those generated by ionising radiation are among the principal causative factors behind mutations, chromosomal aberrations, genetic instability and carcino-genesis, significantly less is known about the epigenetic consequences of double-strand break formation and repair for carcinogenesis. Double-strand break repair is a highly coordinated process that requires the unravelling of the compacted chromatin structure to facilitate repair machinery access and then restoration of the original undamaged chromatin state. Recent experimental findings have pointed to a potential mechanism for double-strand break-induced epigenetic silencing. This review will discuss some of the key epigenetic regulatory processes involved in double-strand break (DSB) repair and how incomplete or incorrect restoration of chromatin structure can leave a DSB-induced epigenetic memory of damage with potentially pathological repercussions

God Bless come back : new experiments with nostalgia in Indonesian rock

This article discusses the production of an Indonesian rock past through a case study of the 1970s rock band God Bless, which has been gradually ‘coming back’ since the middle of the 2000s. In doing so, the article documents this comeback, analyses shifts in the band’s position vis-à-vis nationality, and places these shifts in the context of the industrial and aesthetic transformation of Indonesian popular music over the past decade or so. Furthermore, it considers how the range of nostalgic productions associated with the comeback might be understood not only in light of the scholarship on nostalgia, but also the political environment it inhabits.

Teacher education for high-poverty schools : keeping the bar high

For some time now, there has been a focus, both in Australia and internationally, on quality teaching as a fundamental component that affects the educational outcomes of all students. The question of how teacher education programs in Australia prepare effective teachers to work across all school settings-including low-SES schools-has been elevated to national prominence by data from the 20 12 Programme for International Student Assessment (PIS A), which revealed a fall in Australian students' world ranking across Mathematics, Reading and Science. Education is commonly acknowledged as a "foundation capability" that improves a person's life chances, including employment prospects, and it is widely understood to be a "route out of disadvantage" (McLachlan, Gilfillan, and Gordon 20 13). The Australian Bureau of Statistics 201 1- 12 data suggest that around 2.6 million (11.8%) Australians currently live under the poverty line (Phillips et a!. 2012, 8). According to the Organisation for Economic Cooperation and Development (OECD), despite the significant effects teachers have on student performance, disadvantaged schools are not always staffed with the highest quality teachers (see Darling-Hammond, 2006).

Visual rehabilitation in low-moderate keratoconus : intracorneal ring segment implantation followed by same-day topography-guided photorefractive keratectomy and collagen cross linking

AIM: To present the results of same-day topography-guided photorefractive keratectomy (TG-PRK) and corneal collagen crosslinking (CXL) after previous intrastromal corneal ring segment (ISCR) implantation for keratoconus. METHODS: An experimental clinical study on twenty-one eyes of 19 patients aged, 27.1±6.6 years (range: 19 – 43 years), with low to moderate keratoconus who were selected to undergo customized TG-PRK immediately followed by same-day CXL, 9 months after ISCR implantation in a university ophthalmology clinic. Refraction, uncorrected (UDVA) and corrected distance visual acuities (CDVA), keratometry (K) values, central corneal thickness (CCT) and coma were assessed 3 months after TG/PRK and CXL. RESULTS: After TG-PRK/CXL: the mean UDVA (logMAR) improved significantly from 0.66±0.41 to 0.20±0.25 (P<0.05); K flat value decreased from: 48.44±3.66 D to 43.71±1.95 D; K steep value decreased from 45.61±2.40 D to 41.56±2.05D; K average also decreased from 42.42±2.07 D to 47.00±2.66 D (P<0.05 for all). The mean sphere and cylinder decreased significantly post-surgery from, -3.10±2.99 D to -0.11±0.93 D and from, -3.68±1.53 to -1.11±0.75D respectively, while the CDVA, CCT and coma showed no significant changes. Compared to post-ISCR, significant reductions (P ˂ 0.05 or all) in all K-values, sphere and cylinder were observed after TG-PRK/CXL. CONCLUSION: Same-day combined topography-guided PRK and corneal crosslinking following placement of ICRS is a safe and potentially effective option in treating low-moderate keratoconus. It significantly improved all visual acuity, reduced keratometry, sphere and astigmatism, but caused no change in central corneal thickness and coma.

Computational epigenetic profiling of CpG islets in MTHFR

Computational epigenetics is a new area of research focused on exploring how DNA methylation patterns affect transcription factor binding that affect gene expression patterns. The aim of this study was to produce a new protocol for the detection of DNA methylation patterns using computational analysis which can be further confirmed by bisulfite PCR with serial pyrosequencing. The upstream regulatory element and pre-initiation complex relative to CpG islets within the methylenetetrahydrofolate reductase gene were determined via computational analysis and online databases. The 1,104 bp long CpG island located near to or at the alternative promoter site of methylenetetrahydrofolate reductase gene was identified. The CpG plot indicated that CpG islets A and B, within the island, contained 62 and 75 % GC content CpG ratios of 0.70 and 0.80–0.95, respectively. Further exploration of the CpG islets A and B indicates that the transcription start sites were GGC which were absent from the TATA boxes. In addition, although six PROSITE motifs were identified in CpG B, no motifs were detected in CpG A. A number of cis-regulatory elements were found in different regions within the CpGs A and B. Transcription factors were predicted to bind to CpGs A and B with varying affinities depending on the DNA methylation status. In addition, transcription factor binding may influence the expression patterns of the methylenetetrahydrofolate reductase gene by recruiting chromatin condensation inducing factors. These results have significant implications for the understanding of the architecture of transcription factor binding at CpG islets as well as DNA methylation patterns that affect chromatin structure.

Butterfly catastrophe for fronts in a three-component reaction–diffusion system

We study the dynamics of front solutions in a three-component reaction–diffusion system via a combination of geometric singular perturbation theory, Evans function analysis, and center manifold reduction. The reduced system exhibits a surprisingly complicated bifurcation structure including a butterfly catastrophe. Our results shed light on numerically observed accelerations and oscillations and pave the way for the analysis of front interactions in a parameter regime where the essential spectrum of a single front approaches the imaginary axis asymptotically.

The influence of internal communication on employee engagement: A pilot study

Internal communication is an important concept and integral to internal public relations with links to positive organizational and employee outcomes such as employee engagement. While scholars emphasize the importance of the relationship between internal communication and employee engagement, the association has not been empirically tested. Using surveys and regression analysis this study confirms that internal organizational communication and internal supervisor communication support workplace relationships based on meaning and worth, and have a significant part to play in developing and maintaining optimal employee engagement.

Experimental Study of crack propagation in carbon steel using acoustic emission

Acoustic emission technique has become a significant and powerful structural health monitoring tool for structures. Researches to date have been done on crack location, fatigue crack propagation in materials and severity assessment of failure using acoustic emission technique. Determining severity of failure in steel structures using acoustic emission technique is still a challenge to accurately determine the relationship between the severity of crack propagation and acoustic emission activities. In this study three point bending test on low carbon steel samples along with acoustic emission technique have been used to determine crack propagation and severity. A notch is introduced at the tension face of the loading point to the samples to initiate the crack. The results show that the percentage of load drop of the steel specimen has a reciprocal relationship with the crack opening i.e. crack opening zones are influenced by the loading rate. In post yielding region, common acoustic emission signal parameters such as, signal strength, energy and amplitudes are found to be higher than those at pre-yielding and at yielding.