Functional interaction between Epstein-Barr virus replication protein Zta and host DNA damage response protein 53BP1

Epstein-Barr virus (EBV; human herpesvirus 4) poses major clinical problems worldwide. Following primary infection, EBV enters a form of long-lived latency in B lymphocytes, expressing few viral genes, and it persists for the lifetime of the host with sporadic bursts of viral replication. The switch between latency and replication is governed by the action of a multifunctional viral protein Zta (also called BZLF1, ZEBRA, and Z). Using a global proteomic approach, we identified a host DNA damage repair protein that specifically interacts with Zta: 53BP1. 53BP1 is intimately connected with the ATM signal transduction pathway, which is activated during EBV replication. The interaction of 53BP1 with Zta requires the C-terminal ends of both proteins. A series of Zta mutants that show a wild-type ability to perform basic functions of Zta, such as dimer formation, interaction with DNA, and the transactivation of viral genes, were shown to have lost the ability to induce the viral lytic cycle. Each of these mutants also is compromised in the C-terminal region for interaction with 53BP1. In addition, the knockdown of 53BP1 expression reduced viral replication, suggesting that the association between Zta and 53BP1 is involved in the viral replication cycle

After hours nurse staffing, work intensity and quality of care - Missed Care Study: South Australia

During November, 2012, the Flinders University After Hours Nurse Staffing, Work Intensity and Quality of Care project team, in collaboration with the Australian Nursing and Midwifery Federation, SA Branch (ANMFSA), administered the MISSCARE survey to a sample of 354 nurse/midwife members of ANMFSA. The survey contained 13 demographic questions, 28 questions that explored working conditions, 96 questions concerning missed nursing care (defined as care that is omitted, postponed, or incomplete) and 17 questions concerning perceived reasons care is omitted in the settings in which the nurse/midwives practice. In addition, respondents were asked to add comments of their own concerning nursing care that is missed and why

Nurses and midwives perceptions of missed nursing care – A South Australian study

Author version made available in accordance with the publisher's policy for non-mandated open access submission. Under Elsevier's copyright, non-mandated authors are permitted to make work available in an institutional repository. NOTICE: this is the author’s version of a work that was accepted for publication in Collegian. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in COLLEGIAN, [2014] DOI:10.1016/ j.colegn.2014.09.001Background Budgetary restrictions and shorter hospital admission times have increased demands upon nursing time leading to nurses missing or rationing care. Previous research studies involving perceptions of missed care have predominantly occurred outside of Australia. This paper reports findings from the first South Australian study to explore missed nursing care. Aim To determine and explore nurses’ perceptions of reasons for missed care within the South Australian context and across a variety of healthcare settings. Method The survey was a collaborative venture between the Flinders University of South Australia, After Hours Nurse Staffing Work Intensity and Quality of Care project team and the Australian Nursing and Midwifery Federation, SA Branch. Electronic invitations using Survey Monkey were sent to randomly selected nurses and midwives and available online for two months. Three hundred and fifty four nurses and midwives responded. Recurring issues were identified from qualitative data within the survey and three main reasons for missed care emerged. Findings Three main reasons for missed care were determined as: competing demands that reduce time for patient care; ineffective methods for determining staffing levels; and skill mix including inadequate staff numbers. These broad issues represented respondents’ perceptions of missed care. Conclusion Issues around staffing levels, skill mix and the ability to predict workload play a major role in the delivery of care. This study identified the increasing work demands on nurses/midwifes. Solutions to the rationing of care need further exploration

Methylated DNA recognition during the reversal of epigenetic silencing is regulated by cysteine and cerine residues in the Epstein-Barr Virus lytic switch protein

Epstein-Barr virus (EBV) causes infectious mononucleosis and is associated with various malignancies, including Burkitt's lymphoma and nasopharyngeal carcinoma. Like all herpesviruses, the EBV life cycle alternates between latency and lytic replication. During latency, the viral genome is largely silenced by host-driven methylation of CpG motifs and, in the switch to the lytic cycle, this epigenetic silencing is overturned. A key event is the activation of the viral BRLF1 gene by the immediate-early protein Zta. Zta is a bZIP transcription factor that preferentially binds to specific response elements (ZREs) in the BRLF1 promoter (Rp) when these elements are methylated. Zta's ability to trigger lytic cycle activation is severely compromised when a cysteine residue in its bZIP domain is mutated to serine (C189S), but the molecular basis for this effect is unknown. Here we show that the C189S mutant is defective for activating Rp in a Burkitt's lymphoma cell line. The mutant is compromised both in vitro and in vivo for binding two methylated ZREs in Rp (ZRE2 and ZRE3), although the effect is striking only for ZRE3. Molecular modeling of Zta bound to methylated ZRE3, together with biochemical data, indicate that C189 directly contacts one of the two methyl cytosines within a specific CpG motif. The motif's second methyl cytosine (on the complementary DNA strand) is predicted to contact S186, a residue known to regulate methyl-ZRE recognition. Our results suggest that C189 regulates the enhanced interaction of Zta with methylated DNA in overturning the epigenetic control of viral latency. As C189 is conserved in many bZIP proteins, the selectivity of Zta for methylated DNA may be a paradigm for a more general phenomenon

Brain Morphometry and Cognitive Outcomes in Adolescents and Adults with Complex Congenital Heart Disease

This thesis examines long-term cognitive outcomes and brain morphometry in adolescents and adults with complex CHD. Consideration is given to the broader impact of these challenges on psychosocial outcomes, and we attempt to advance our understanding of potentially modifiable risk factors. Strategies to address the insufficiencies in current clinical resources are explored. In Chapter 3 we demonstrate that young people with a Fontan circulation experience marked reductions in cognitive functioning in comparison to age- and sex-matched individuals with transposition of the great arteries and healthy controls. Structural brain injury was not indicative of worse cognitive outcomes. Whereas global brain volumes appear to be a key structural underpin contributing to cognitive dysfunction and are associated with lower resting oxygen saturations. Chapter 4 advances these findings and demonstrates that reduced resting and peak exercise oxygen saturations are significantly associated with worse white matter microstructural integrity, indicating that persisting hypoxaemia may be a modifiable risk factor contributing to long-term brain abnormality. In Chapter 5 we show that lower socioeconomic status is a risk factor for worse cognitive outcomes in adults with complex CHD, who may not recognise or report their cognitive challenges. Psychological distress is common though was not a strong correlate of cognitive functioning. Chapter 6 demonstrates that Cogstate may be a useful screening tool to identify individuals with CHD who are most in need of a formal evaluation. Combined, this work adopts the biopsychosocial approach to wellness and investigates biological, psychological, and social factors that contribute to cognitive dysfunction in complex CHD. The research serves to guide future clinical practice and the development of theory driven intervention strategies to optimise long-term outcomes for those at risk

The reversal of epigenetic silencing of the EBV genome is regulated by viral bZIP protein

EBV (Epstein-Barr virus) alternates between latency and lytic replication. During latency, the viral genome is largely silenced by host-driven methylation of CpG motifs and in the switch to the lytic cycle this epigenetic silencing is overturned. A key event is the activation of the viral protein Zta with three ZREs (Zta-response elements) from the BRLF1 promoter (referred to as Rp). Two of these ZREs contain CpG motifs and are methylated in the latent genome. Biochemical analyses and molecular modelling of Zta bound to methylated RpZRE3 indicate the precise contacts made between a serine and a cysteine residue of Zta with methyl cytosines. A single point mutant of Zta, C189S, is defective in binding to the methylated ZREs both in vitro and in vivo. This was used to probe the functional relevance of the interaction. ZtaC189S was not able to activate Rp in a B-cell line, demonstrating the relevance of the interaction with methylated ZREs. This demonstrates that Zta plays a role in overturning the epigenetic control of viral latency. © The Authors Journal compilation © 2008 Biochemical Society

Mapping social processes at work in nursing knowledge development

Harvey, CL ORCiD: 0000-0001-9016-8840In this paper, we suggest a blueprint for combining bibliometrics and critical analysis as a way to review published scientific works in nursing. This new approach is neither a systematic review nor meta-analysis. Instead, it is a way for researchers and clinicians to understand how and why current nursing knowledge developed as it did. Our approach will enable consumers and producers of nursing knowledge to recognize and take into account the social processes involved in the development, evaluation, and utilization of new nursing knowledge. We offer a rationale and a strategy for examining the socially-sanctioned actions by which nurse scientists signal to readers the boundaries of their thinking about a problem, the roots of their ideas, and the significance of their work. These actions - based on social processes of authority, credibility, and prestige - have bearing on the careers of nurse scientists and on the ways the knowledge they create enters into the everyday world of nurse clinicians and determines their actions at the bedside, as well as their opportunities for advancement. © 2014 Wiley Publishing Asia Pty Ltd

Priced to care: Factors underpinning missed care

Harvey, CL ORCiD: 0000-0001-9016-8840This article examines missed care through dialogues examining the perceptions of nurses in regard to missed care occasions. Using a critical discourse analysis (CDA), the study explores the truth claims of participants who describe the challenges they encounter in daily attempts to deliver what they consider effective patient care. These are compared to the mandates of state and organisational policy prescribing clinical practice. The boundaries of tension that are expressed by nurses within the milieu of missed care are explored through in-depth interviews. CDA is interested in social organisation and the interplay of people's activities within it, the focus being on how they construe and internalise such activity. Nurses' perceptions and realities become central to any investigation because they are often organised by more than their own intentions or motivations, with influences such as professional standards or organisational rules subconsciously locating their reality. Instead of identifying occasions of omitted care, nurses spoke of constraints related to budget, staffing, skill mix and mandated policy as constraining their ability to complete care activities. Factors emerged that suggest that missed care is the consequence of routinised and standardised practice, cited as cost effective care, at the expense of professional autonomy. © 2016, © Australian Labour and Employment Relations Association (ALERA), SAGE Publications Ltd, Los Angeles, London, New Delhi, Singapore and Washington DC

Singlet oxygen production by UV and visible photoexcited porphyrins under different states of aggregation

peer reviewedTryptophan photosensitization was used to determine the quantum yield of singlet oxygen production by porphyrins irradiated with ultraviolet or visible light in phosphate buffer at pH 7.0. From a preliminary study of aggregation of these dyes, it was established that uroporphyrin, hematoporphyrin and deuteroporphyrin allowed to investigate the behaviour of monomers, dimers and higher aggregates, respectively. The photosensitization results show that monomers and polymers produce singlet oxygen with very similar yields. Inclusion of Zn2+ and Cu2+ metal ions on deuteroporphyrin inhibits partially (factor 3) and totally the singlet oxygen production