N-terminal acetylome analyses and functional insights of the N-terminal acetyltransferase NatB

Protein N-terminal acetylation (Nt-acetylation) is an important mediator of protein function, stability, sorting, and localization. Although the responsible enzymes are thought to be fairly well characterized, the lack of identified in vivo substrates, the occurrence of Nt-acetylation substrates displaying yet uncharacterized N-terminal acetyltransferase (NAT) specificities, and emerging evidence of posttranslational Nt-acetylation, necessitate the use of genetic models and quantitative proteomics. NatB, which targets Met-Glu-, Met-Asp-, and Met-Asn-starting protein N termini, is presumed to Nt-acetylate 15% of all yeast and 18% of all human proteins. We here report on the evolutionary traits of NatB from yeast to human and demonstrate that ectopically expressed hNatB in a yNatB-Δ yeast strain partially complements the natB-Δ phenotypes and partially restores the yNatB Nt-acetylome. Overall, combining quantitative N-terminomics with yeast studies and knockdown of hNatB in human cell lines, led to the unambiguous identification of 180 human and 110 yeast NatB substrates. Interestingly, these substrates included Met-Gln- N-termini, which are thus now classified as in vivo NatB substrates. We also demonstrate the requirement of hNatB activity for maintaining the structure and function of actomyosin fibers and for proper cellular migration. In addition, expression of tropomyosin-1 restored the altered focal adhesions and cellular migration defects observed in hNatB-depleted HeLa cells, indicative for the conserved link between NatB, tropomyosin, and actin cable function from yeast to human.

Comparing different approach and avoidance models of learning and personality in the prediction of work, university, and leadership outcomes

Jackson (2005) developed a hybrid model of personality and learning, known as the learning styles profiler (LSP) which was designed to span biological, socio-cognitive, and experiential research foci of personality and learning research. The hybrid model argues that functional and dysfunctional learning outcomes can be best understood in terms of how cognitions and experiences control, discipline, and re-express the biologically based scale of sensation-seeking. In two studies with part-time workers undertaking tertiary education (N equals 137 and 58), established models of approach and avoidance from each of the three different research foci were compared with Jackson's hybrid model in their predictiveness of leadership, work, and university outcomes using self-report and supervisor ratings. Results showed that the hybrid model was generally optimal and, as hypothesized, that goal orientation was a mediator of sensation-seeking on outcomes (work performance, university performance, leader behaviours, and counterproductive work behaviour). Our studies suggest that the hybrid model has considerable promise as a predictor of work and educational outcomes as well as dysfunctional outcomes.

In conversation with the Honourable Ian Callinan AC

When, in 1977, the Australian electorate provided a double majority to effect a change of section 72 of the Commonwealth Constitution requiring judges of the High Court of Australia to retire at the age of 70 years old, I doubt we understood the continuing capacity of these esteemed members of the judiciary. For the opportunity to sit and talk with Ian Callinan AC who, in compliance with that amendment, retired from the High Court in September 2007, I needed to wait until he returned from The Hague where he was sitting as a Judge ad hoc on the International Court of Justice. Although a native of Casino, New South Wales, Mr Callinan is regarded as a Queenslander. Indeed, he grew up in Brisbane, finished high school at Brisbane Grammar and graduated in law at The University of Queensland. Appointed in 1978 as a Queen’s Counsel, Mr Callinan enjoyed this period of his legal career and we discussed an aspect of the Christopher Skase case, which reinforced my belief that Mr Callinan is an incredibly skilful advocate. On 14 September 1998, ABC Four Corners broadcasted the views of some prominent Australians on the appointment of Mr Callinan to the High Court. In assessing the type of person Mr Callinan is, Tony Morris QC said: “Ian Callinan isn't a coward”, while former Commonwealth Attorney-General, Michael Lavarch, said: “He was regarded as an absolutely outstanding criminal lawyer within the Queensland legal profession, I mean really a top-notch advocate”. I was not interested in raising any of the controversial issues that Mr Callinan has encountered as an advocate in high profile matters. I wanted to know how he felt about his time on the High Court, what his thoughts are on the operation of the High Court, the IP cases he decided, the real life issues that he feels impact on counsel who are appearing before the High Court and the people he regarded as role models. During our conversation, Mr Callinan laughed often and when he did his eyes lit up, revealing his passion for life. He is an incredibly genuine Australian who loved his time as a barrister, enjoyed his role on the High Court, enjoys his current job as mediator, loves writing novels, has a great desire for continual improvement in the quality of legal education and legal advocacy and sees a need for change in IP law. When I asked: “So, what might the future hold for you?”, he laughed and said: “Well, at my age I don’t have a long horizon time”. I said: “Just enjoy the journey?”, to which Mr Callinan responded: “Exactly”.

What predicts mental health in midlife and older women? Results from the Australian Healthy Aging of Women Study

Objectives: Previous research has linked unhealthy lifestyle with a range of negative health outcomes in women. As women age however, they may have fewer performance expectations, but may view their health more positively. Clearly, the experiences of midlife and older women in relation to health and wellbeing need further exploration. The purpose of this study is to examine the factors associated with poor health-related quality of life in midlife (HRQoL) and older Australian women. Methods: The Australian longitudinal Healthy Aging of Women (HOW) study prospectively examines HRQoL, chronic disease and modifiable lifestyle factors midlife and older women as they age. Random sampling was used to select rural and urban based women from South-East Queensland, Australia. Data were collected from 386 women at three time points over the last decade (2001, 2004 and 2011). Results: The average age of women in this study was 65 years (SD = 2.82). Almost three-quarters (73%, n = 248) of the sample were married or living as though married, nine per cent (n = 30) were separated or divorced and a small proportion were had never married (n = 13). Most (86%, n = 291) of the women sample reported being Australian born, around one quarter (34%, n = 114) had completed additional study since leaving school (university degree or diploma). Over half (55%, n = 186) of participants were retired, one quarter (25%, n = 85) were in paid employment and the remained were unemployed (1%, n = 4), unable to work because of illness (2%, n = 6) or worked within the home (17%, n = 56). Using data collected over time we examined the relationship between a range of modifiable lifestyle factors and mental health using structural equation modelling. The overall model exhibited a good fit with the data. Poor sleep quality was associated with reduced mental health while better mental health was reported in women who exercised regularly and satisfied with their currently weight. As hypothesized, past mental health was a significant mediator of current mental health. Conclusions: These findings demonstrate that the mental health of women is complex and needs to be understood not only in terms of current lifestyle but also in relation to previously reported health status.

Unpacking the role of mindfulness on conscientiousness and spirituality

The present study examined relationships between conscientiousness and intrinsic spirituality, with the proposed trait ‘mindfulness’ as mediator. The results from 161 functioning adults within an Australian context revealed that mindfulness was significantly predicted by conscientiousness. This study examined the relationship among conscientiousness, trait mindfulness and intrinsic spirituality. It was hypothesised that trait mindfulness would mediate the relationship between conscientiousness and spirituality. We found this hypothesis partially supported. Practically, these results suggest that conscientious individuals do significantly connect with mindfulness, and it was only the more mindful of conscientious individuals that also displayed high levels of intrinsic spirituality. Additional analyses also suggest that conscientious individuals connect with mindfulness through attending to current actions or regulating impulses (act aware) and have an accepting attitude towards thoughts and feelings (non-judge). Possible explanations and implications of these results are discussed in relation to theory, practice and delivery mechanisms of mindfulness.

Integrin-linked kinase as a target for ERG-mediated invasive properties in prostate cancer models

Approximately half of prostate cancers (PCa) carry TMPRSS2-ERG translocations; however, the clinical impact of this genomic alteration remains enigmatic. Expression of v-ets erythroblastosis virus E26 oncogene like (avian) gene (ERG) promotes prostatic epithelial dysplasia in transgenic mice and acquisition of epithelial-to-mesenchymal transition (EMT) characteristics in human prostatic epithelial cells (PrECs). To explore whether ERG-induced EMT in PrECs was associated with therapeutically targetable transformation characteristics, we established stable populations of BPH-1, PNT1B and RWPE-1 immortalized human PrEC lines that constitutively express flag-tagged ERG3 (fERG). All fERG-expressing populations exhibited characteristics of in vitro and in vivo transformation. Microarray analysis revealed >2000 commonly dysregulated genes in the fERG-PrEC lines. Functional analysis revealed evidence that fERG cells underwent EMT and acquired invasive characteristics. The fERG-induced EMT transcript signature was exemplified by suppressed expression of E-cadherin and keratins 5, 8, 14 and 18; elevated expression of N-cadherin, N-cadherin 2 and vimentin, and of the EMT transcriptional regulators Snail, Zeb1 and Zeb2, and lymphoid enhancer-binding factor-1 (LEF-1). In BPH-1 and RWPE-1-fERG cells, fERG expression is correlated with increased expression of integrin-linked kinase (ILK) and its downstream effectors Snail and LEF-1. Interfering RNA suppression of ERG decreased expression of ILK, Snail and LEF-1, whereas small interfering RNA suppression of ILK did not alter fERG expression. Interfering RNA suppression of ERG or ILK impaired fERG-PrEC Matrigel invasion. Treating fERG-BPH-1 cells with the small molecule ILK inhibitor, QLT-0267, resulted in dose-dependent suppression of Snail and LEF-1 expression, Matrigel invasion and reversion of anchorage-independent growth. These results suggest that ILK is a therapeutically targetable mediator of ERG-induced EMT and transformation in PCa.

The hunt

"Information Thru Play: In 2010, responding to the success of The Threshold, Juxt Interactive again asked No Mimes Media, to partner in creating a transmedia experience to entertain and inform Cisco's Global Sales Force. The Hunt put employees at the center of a thriller where characters sent and responded to their emails, left phone messages, communicated through Facebook and Twitter, even asked them to retrieve items from a dead drop and to send them photographs and information. And while helping fictional characters Isabel and Keith escape an ancient secret organization, the sales force also learned about new Cisco technologies coming to market. Cisco had new demands for the 2010 experience. A geographically and culturally dispersed sales force raises challenges when it comes to introducing dozens of new products and technologies each year. Cisco wanted The Hunt to have global reach, to educate, to build collaboration, and to be fun. This demanded new ways of storytelling and new ways of thinking. The Hunt was quick and intense, unfolding in real time in just two weeks. Many experienced players were poised to participate and expectations were high. Many of the mechanics of the previous year's experience were repeated, and the audience ripped through the opening, discovering video clips and websites in minutes. The surprise was discovering Facebook and Twitter accounts, where characters responded to player postings and comments in real time. The Hunt involved audience members from countries around the world, including China, India, Netherlands, Germany, Norway, Pakistan, Japan, the United Kingdom, and the United States. It highlighted new Cisco technologies like Pulse and Mediator, painlessly engaging the audience in what those technologies do and how they work. Players collaborated across silos, creating networks of cross-disciplinary experts. The Hunt pushed the boundaries of storytelling, with events unfolding on Twitter and Facebook, and in the real world where the audience had to use social engineering to find and secure a package with vital information. With thousands of players highly engaged around the world, The Hunt once again proved that transmedia experiences can effectively be used to not only meet the goals of a brand, but entertain their audience as well."

DNA methylation at the novel CpG sites in the promoter of MED15/PCQAP gene as a biomarker for head and neck cancers

Head and neck cancers (HNCs) represent a significant and ever-growing burden to the modern society, mainly due to the lack of early diagnostic methods. A significant number of HNCs is often associated with drinking, smoking, chewing beetle nut, and human papilloma virus (HPV) infections. We have analyzed DNA methylation patterns in tumor and normal tissue samples collected from head and neck squamous cell carcinoma (HNSCC) patients who were smokers. We have identified novel methylation sites in the promoter of the mediator complex subunit 15 (MED15/PCQAP) gene (encoing a co-factor important for regulation of transcription initiation for promoters of many genes), hypermethylated specifically in tumor cells. Two clusters of CpG dinucleotides methylated in tumors, but not in normal tissue from the same patients, were identified. These CpG methylation events in saliva samples were further validated in a separate cohort of HNSCC patients (who developed cancer due to smoking or HPV infections) and healthy controls using methylation-specific PCR (MSP). We used saliva as a biological medium because of its non-invasive nature, close proximity to the tumors, easiness and it is an economically viable option for large-scale screening studies. The methylation levels for the two identified CpG clusters were significantly different between the saliva samples collected from healthy controls and HNSCC individuals (Welch's t-test returning P, 0.05 and Mann-Whitney test P, 0.01 for both). The developed MSP assays also provided a good discriminative ability with AUC values of 0.70 (P, 0.01) and 0.63 (P, 0.05). The identified novel CpG methylation sites may serve as potential non-invasive biomarkers for detecting HNSCC. © the authors.

The MyD88+ phenotype is an adverse prognostic factor in epithelial ovarian cancer

The prognosis of epithelial ovarian cancer is poor in part due to the high frequency of chemoresistance. Recent evidence points to the Toll-like receptor-4 (TLR4), and particularly its adaptor protein MyD88, as one potential mediator of this resistance. This study aims to provide further evidence that MyD88 positive cancer cells are clinically significant, stem-like and reproducibly detectable for the purposes of prognostic stratification. Expression of TLR4 and MyD88 was assessed immunohistochemically in 198 paraffin-embedded ovarian tissues and in an embryonal carcinoma model of cancer stemness. In parallel, expression of TLR4 and MyD88 mRNA and regulatory microRNAs (miR-21 and miR-146a) was assessed, as well as in a series of chemosensitive and resistant cancer cells lines. Functional analysis of the pathway was assessed in chemoresistant SKOV-3 ovarian cancer cells. TLR4 and MyD88 expression can be reproducibly assessed via immunohistochemistry using a semi-quantitative scoring system. TLR4 expression was present in all ovarian epithelium (normal and neoplastic), whereas MyD88 was restricted to neoplastic cells, independent of tumour grade and associated with reduced progression-free and overall survival, in an immunohistological specific subset of serous carcinomas, p<0.05. MiR-21 and miR-146a expression was significantly increased in MyD88 negative cancers (p<0.05), indicating their participation in regulation. Significant alterations in MyD88 mRNA expression were observed between chemosensitive and chemoresistant cells and tissue. Knockdown of TLR4 in SKOV-3 ovarian cells recovered chemosensitivity. Knockdown of MyD88 alone did not. MyD88 expression was down-regulated in differentiated embryonal carcinoma (NTera2) cells, supporting the MyD88+ cancer stem cell hypothesis. Our findings demonstrate that expression of MyD88 is associated with significantly reduced patient survival and altered microRNA levels and suggest an intact/functioning TLR4/MyD88 pathway is required for acquisition of the chemoresistant phenotype. Ex vivo manipulation of ovarian cancer stem cell (CSC) differentiation can decrease MyD88 expression, providing a potentially valuable CSC model for ovarian cancer.

Children's advertising literacy for advergames : perception of the game as advertising

This study examined whether children recognized advergames as a type of advertising and the efficacy of an advertising literacy program. Results indicated that without the advertising literacy education, about three-quarters of the children did not recognize advergames as a type of advertising. However, those with advertising literacy education showed a significantly enhanced understanding. Also, a series of mediation tests showed that recognition of advertising was an indirect-only mediator between the advertising literacy and skeptical attitudes toward advertising. Only those who viewed the advergame as a type of advertising demonstrated more skeptical attitudes toward it.

Electrocatalytic and SERS activity of Pt rich Pt-Pb nanostructures formed via the utilisation of in-situ underpotential deposition of lead

The controlled synthesis of nanostructured materials remains an ongoing area of research, especially as the size, shape and composition of nanomaterials can greatly influence their properties and applications. In this work we present the electrodeposition of highly dendritic platinum rich platinum-lead nanostructures, where lead acetate acts as an inorganic shape directing agent via underpotential deposition on the growing electrodeposit. It was found that these nanomaterials readily oxidise at potentials below monolayer oxide formation, which significantly impacts on the methanol electrooxidation reaction and correlates with the incipient hydrous oxide adatom mediator (IHOAM) model of electrocatalysis. Additionally these materials were tested for their surface enhanced Raman scattering (SERS) activity, where the high density of sharp tips provides promise for their application as SERS substrates.

An SECM study on the influence of cationic, membrane-active peptides on a gold-supported self-assembled monolayer

The influence of the membrane active peptides, Tat44–57 (activator in HIV-1) and melittin (active content of bee venom), on self-assembled monolayers of 6-mercaptohexanoic acid (MHA) on gold electrodes has been studied with scanning electrochemical microscopy (SECM). It was found that MHA, when deprotonated at physiological pH, significantly affected the relative rates of electron transfer between the [Fe(CN)6]4− solution based mediator and the underlying gold electrode, predominantly by the electrostatic interaction between the mediator and MHA. Upon the introduction of Tat44–57 ormelittin to the electrolyte, the relative rate of electron transfer through the MHA layer could be increased or decreased depending on the mediator used. However, in all cases it was found that these peptides have the ability to be incorporated into synthetic SAMs, which has implications for future electrochemical studies carried out using cell mimicking membranes immobilised on such layers.

Childhood atopic dermatitis : a cross-sectional study of relationships between child and parent factors, atopic dermatitis management, and disease severity

Background: Successful management of atopic dermatitis poses a significant and ongoing challenge to parents of affected children. Despite frequent reports of child behaviour problems and parenting difficulties, there is a paucity of literature examining relationships between child behaviour and parents' confidence and competence with treatment. Objectives: To examine relationships between child, parent, and family variables, parents' self-efficacy for managing atopic dermatitis, self-reported performance of management tasks, observed competence with providing treatment, and atopic dermatitis severity. Design: Cross-sectional study design. Participants A sample of 64 parent-child dyads was recruited from the dermatology clinic of a paediatric tertiary referral hospital in Brisbane, Australia. Methods: Parents completed self-report questionnaires examining child behaviour, parents' adjustment, parenting conflict, parents' relationship satisfaction, and parents' self-efficacy and self-reported performance of key management tasks. Severity of atopic dermatitis was assessed using the Scoring Atopic Dermatitis index. A routine home treatment session was observed, and parents' competence in carrying out the child's treatment assessed. Results: Pearson's and Spearman's correlations identified significant relationships (p< .05) between parents' self-efficacy and disease severity, child behaviour difficulties, parent depression and stress, parenting conflict, and relationship satisfaction. There were also significant relationships between each of these variables and parents' self-reported performance of management tasks. More profound child behaviour difficulties were associated with more severe atopic dermatitis and greater parent stress. Using multiple linear regressions, significant proportions of variation in parents' self-efficacy and self-reported task performance were explained by child behaviour difficulties and parents' formal education. Self-efficacy emerged as a likely mediator for relationships between both child behaviour and parents' education, and self-reported task performance. Direct observation of treatment sessions revealed strong relationships between parents' treatment competence and parents' self-efficacy, outcome expectations, and self-reported task performance. Less competent task performance was also associated with greater parent-reported child behaviour difficulties, parent depression and stress, parenting conflict, and relationship dissatisfaction. Conclusion: This study revealed the importance of child behaviour to parents' confidence and practices in the context of atopic dermatitis management. Children with more severe atopic dermatitis are at risk of presenting with challenging behaviour problems and their parents struggle to manage the condition successfully.

The trustworthy brand : effects of conclusion explicitness and persuasion awareness on consumer judgments

Two studies examine how a consumer’s awareness of marketing tactics influences the effectiveness of conclusion explicitness advertising (implicit, open-ended or explicit, closed-ended conclusions). Study 1 shows that persuasion awareness and conclusion explicitness influence brand evaluations. Persuasion aware consumers prefer implicit conclusions in comparative advertising that allow them to decide which brand is superior, rather than explicit conclusions which state the superior brand. Persuasion unaware consumers show no difference for conclusion explicitness. Brand trust mediates the results. Study 2 demonstrates the robustness of these effects. Research contributions include persuasion awareness as a moderator of conclusion explicitness effects and the role of trust as a mediator. For managers, results show how implicit conclusions can improve the brand evaluations of persuasion aware consumers.

Starting with strengths : an Indigenous early years intervention

Aboriginal protocol usually links the right to tell a story with a declaration of involvement or connection to the story. I am Aboriginal . . . I am a woman, daughter, sister, aunty and wife. I am also a mother to three beautiful children aged 6, 4 and 2 years. To my children at this point in their lives, I am their provider, nurturer, teacher, cook, taxi driver, mediator, stylist, Elder, slave, and expert on all there is to know in the world. Being the centre of the universe to three impressionable young minds is a role that I cherish deeply, and I take the responsibilities of it very seriously. I love the job of parenting. As any parent would agree, it is the most challenging and difficult job of all, but the opportunity to bring a life into the world and shape and mould a little person into a big person brings rewards that no career can.