Multi-omics analysis of glucose-mediated signaling by a moonlighting Gß protein Asc1/RACK1.

Heterotrimeric G proteins were originally discovered through efforts to understand the effects of hormones, such as glucagon and epinephrine, on glucose metabolism. On the other hand, many cellular metabolites, including glucose, serve as ligands for G protein-coupled receptors. Here we investigate the consequences of glucose-mediated receptor signaling, and in particular the role of a Gα subunit Gpa2 and a non-canonical Gß subunit, known as Asc1 in yeast and RACK1 in animals. Asc1/RACK1 is of particular interest because it has multiple, seemingly unrelated, functions in the cell. The existence of such "moonlighting" operations has complicated the determination of phenotype from genotype. Through a comparative analysis of individual gene deletion mutants, and by integrating transcriptomics and metabolomics measurements, we have determined the relative contributions of the Gα and Gß protein subunits to glucose-initiated processes in yeast. We determined that Gpa2 is primarily involved in regulating carbohydrate metabolism while Asc1 is primarily involved in amino acid metabolism. Both proteins are involved in regulating purine metabolism. Of the two subunits, Gpa2 regulates a greater number of gene transcripts and was particularly important in determining the amplitude of response to glucose addition. We conclude that the two G protein subunits regulate distinct but complementary processes downstream of the glucose-sensing receptor, as well as processes that lead ultimately to changes in cell growth and metabolism.

Setting foot in private spaces: extending the hepatitis C cascade of care to automatic needle/syringe dispensing machines, a mixed methods study.

BACKGROUND: Global commitment to achieving hepatitis C virus (HCV) elimination has enhanced efforts in improving access to direct-acting antiviral (DAA) treatments for people who inject drugs (PWID). Scale-up of efforts to engage hard-to-reach groups of PWID in HCV testing and treatment is crucial to success. Automatic needle/syringe dispensing machines (ADMs) have been used internationally to distribute sterile injecting equipment. ADMs are a unique harm reduction service, affording maximum anonymity to service users. This paper explores the feasibility and acceptability of extending the HCV cascade of care to sites where ADMs are located. METHODS: The ADM users into Treatment (ADMiT) study was undertaken in a metropolitan region in Sydney, Australia. This mixed methods study involved analysis of closed-circuit television footage, ethnographic methods (fieldwork observation and in-depth interviews) and structured surveys. Researchers and peers conducted fieldwork and data collection over 10 weeks at one ADM site, including offering access to HCV testing and treatment. RESULTS: Findings from 10 weeks of fieldwork observations, 70 survey participants and 15 interviews highlighted that there is scope for engaging with this population at the time they use the ADM, and enhanced linkage to HCV testing and treatment may be warranted. Most survey participants reported prior HCV testing, 61% in the last 12 months and 38% had received HCV treatment. However, fieldwork revealed that most people observed using the ADM were not willing to engage with the researchers. Field work data and interviews suggested that extending the HCV cascade of care to ADMs may encroach on what is a private space for many PWID, utilized specifically to avoid engagement. DISCUSSION: Enhanced linkage to HCV testing and treatment for people who use ADMs may be warranted. However, data suggested that extending the HCV cascade of care to ADMs may encroach on what is a private space for many PWID, utilized specifically to avoid engagement. The current study raises important public health questions about the need to ensure interventions reflect the needs of affected communities, including their right to remain anonymous.

Career optimism of culturally and linguistically diverse hotel workers in the pandemic age.

The impact of COVID-19 on workers in accommodation and food services-which account for a large proportion of the hospitality sector-is severe, with more than one third of these workers losing their jobs. Against this backdrop, there is heightened interest in the concept of career optimism, yet a dearth of empirical research. Our study addresses this gap by investigating the factors that influence the career optimism of culturally and linguistically diverse (CALD) hotel workers. Using a qualitative study, we found a temporal facet of career optimism for CALD workers, and developed a model to show the individual and contextual factors that enabled and inhibited the career optimism of this group. We thus advance hospitality literature on the careers of CALD workers in hotels and offer practical implications to help CALD hospitality workers stay vocationally optimistic during crisis events such as COVID-19.

A survey of Sydney general practitioners' management of patients with chronic hepatitis B.

OBJECTIVE: To examine the chronic hepatitis B (CHB) assessment and management practices of general practitioners in the Sydney and South Western Sydney Local Health Districts, areas with a high prevalence of CHB, and to obtain their views on alternative models of care. DESIGN, SETTING AND PARTICIPANTS: We used a descriptive, cross-sectional study design to survey GPs who had seen at least one patient aged 18 years or over who had been notified as having CHB to the Public Health Unit between 1 June 2012 and 31 May 2013. There were 213 eligible GPs; the response rate was 57.7%. MAIN OUTCOME MEASURES: The CHB assessment, management and referral practices of the GPs, and their opinions about different models of care. RESULTS: Most GPs (78.9%) were at least reasonably confident about managing CHB. GPs were generally most comfortable with a model of care that involved initial referral to a specialist; managing CHB without specialist input or with only review by a specialised nurse practitioner were less popular. CONCLUSION: These results suggest that barriers, including dependence on specialist input, still hinder the appropriate assessment and management of CHB patients by GPs. Well designed and targeted support programs that include specialist support are needed if there is to be a successful shift to an increased role for GPs in the model of care for managing CHB.

Senescent human fibroblasts show increased glycolysis and redox homeostasis with extracellular metabolomes that overlap with those of irreparable DNA damage, aging, and disease.

Cellular senescence can modulate various pathologies and is associated with irreparable DNA double-strand breaks (IrrDSBs). Extracellular senescence metabolomes (ESMs) were generated from fibroblasts rendered senescent by proliferative exhaustion (PEsen) or 20 Gy of γ rays (IrrDSBsen) and compared with those of young proliferating cells, confluent cells, quiescent cells, and cells exposed to repairable levels of DNA damage to identify novel noninvasive markers of senescent cells. ESMs of PEsen and IrrDSBsen overlapped and showed increased levels of citrate, molecules involved in oxidative stress, a sterol, monohydroxylipids, tryptophan metabolism, phospholipid, and nucleotide catabolism, as well as reduced levels of dipeptides containing branched chain amino acids. The ESM overlaps with the aging and disease body fluid metabolomes, supporting their utility in the noninvasive detection of human senescent cells in vivo and by implication the detection of a variety of human pathologies. Intracellular metabolites of senescent cells showed a relative increase in glycolysis, gluconeogenesis, the pentose-phosphate pathway, and, consistent with this, pyruvate dehydrogenase kinase transcripts. In contrast, tricarboxylic acid cycle enzyme transcript levels were unchanged and their metabolites were depleted. These results are surprising because glycolysis antagonizes senescence entry but are consistent with established senescent cells entering a state of low oxidative stress.

Differences in intradomain and interdomain motion confer distinct activation properties to structurally similar Gα proteins.

Proteins with similar crystal structures can have dissimilar rates of substrate binding and catalysis. Here we used molecular dynamics simulations and biochemical analysis to determine the role of intradomain and interdomain motions in conferring distinct activation rates to two Gα proteins, Gα(i1) and GPA1. Despite high structural similarity, GPA1 can activate itself without a receptor, whereas Gα(i1) cannot. We found that motions in these proteins vary greatly in type and frequency. Whereas motion is greatest in the Ras domain of Gα(i1), it is greatest in helices αA and αB from the helical domain of GPA1. Using protein chimeras, we show that helix αA from GPA1 is sufficient to confer rapid activation to Gα(i1). Gα(i1) has less intradomain motion than GPA1 and instead displays interdomain displacement resembling that observed in a receptor-heterotrimer crystal complex. Thus, structurally similar proteins can have distinct atomic motions that confer distinct activation mechanisms.

G protein activation without a GEF in the plant kingdom.

Animal heterotrimeric G proteins are activated by guanine nucleotide exchange factors (GEF), typically seven transmembrane receptors that trigger GDP release and subsequent GTP binding. In contrast, the Arabidopsis thaliana G protein (AtGPA1) rapidly activates itself without a GEF and is instead regulated by a seven transmembrane Regulator of G protein Signaling (7TM-RGS) protein that promotes GTP hydrolysis to reset the inactive (GDP-bound) state. It is not known if this unusual activation is a major and constraining part of the evolutionary history of G signaling in eukaryotes. In particular, it is not known if this is an ancestral form or if this mechanism is maintained, and therefore constrained, within the plant kingdom. To determine if this mode of signal regulation is conserved throughout the plant kingdom, we analyzed available plant genomes for G protein signaling components, and we purified individually the plant components encoded in an informative set of plant genomes in order to determine their activation properties in vitro. While the subunits of the heterotrimeric G protein complex are encoded in vascular plant genomes, the 7TM-RGS genes were lost in all investigated grasses. Despite the absence of a Gα-inactivating protein in grasses, all vascular plant Gα proteins examined rapidly released GDP without a receptor and slowly hydrolyzed GTP, indicating that these Gα are self-activating. We showed further that a single amino acid substitution found naturally in grass Gα proteins reduced the Gα-RGS interaction, and this amino acid substitution occurred before the loss of the RGS gene in the grass lineage. Like grasses, non-vascular plants also appear to lack RGS proteins. However, unlike grasses, one representative non-vascular plant Gα showed rapid GTP hydrolysis, likely compensating for the loss of the RGS gene. Our findings, the loss of a regulatory gene and the retention of the "self-activating" trait, indicate the existence of divergent Gα regulatory mechanisms in the plant kingdom. In the grasses, purifying selection on the regulatory gene was lost after the physical decoupling of the RGS protein and its cognate Gα partner. More broadly these findings show extreme divergence in Gα activation and regulation that played a critical role in the evolution of G protein signaling pathways.

Feasibility, acceptability and safety of a nurse led hepatitis B clinic based in the community.

AIM: The aim of this study was to investigate if a community based hepatitis B (HBV) nurse clinic is a feasible, acceptable and safe strategy to improve access to best practice chronic hepatitis B care (CHB) in Sydney Local Health District. METHODS: The weekly clinic commenced in an Inner West Sydney Health Centre in November 2012. The CNC responsibilities included patient assessment, management, education, triage, the development of care plans for GPs and GP support. Nursing practice was guided by recommendations from internationally and nationally endorsed CHB Guidelines. Information on patient demographics, clinical findings, triage decisions and sources of referral were collected and used to assess the feasibility, acceptability and safety of the nurse clinic. Patients were also invited to complete a self-administered survey. The survey included questions on attitudes towards the clinic and opinions on barriers to accessing treatment and care. Data was collated and analysed in both Excel and SPPS. RESULTS: In the first 18 months of the clinic 66 people attended, 56 (80%) had CHB, 51 (77%) were born in an Asian country. An equal number of males and females attended. 11 (17%) required further management at a hospital based liver clinic and were referred. 5 (8%) have commenced anti-viral treatment. 24 (36%) met the criteria for six monthly HCC screening and were commenced on HCC surveillance. Twenty-two GPs referred patients. 11 (17%) patients returned the survey and they reported a high level of satisfaction with the clinic and willingness to engage in future CHB care. CONCLUSIONS: This study of a community based CHB nurse clinic shows it is a feasible, acceptable, and safe initiative. The nurse improved access to best practice care and supported patients to effectively manage their CHB. We have confirmed a nurse can have a central role in triage, case management and GP support. Given the high CHB prevalence in our LHD a higher number of GP referrals were expected. Further research on how to increase engagement with GPs and people living with CHB is needed. We plan to expand our model with the CHB nurse conducting assessments and education in GP practices.

Screening patients in general practice for advanced chronic liver disease using an innovative IT solution: The Liver Toolkit.

BACKGROUND: Identifying patients with undiagnosed advanced chronic liver disease (ACLD) is a public health challenge. Patients with advanced fibrosis or compensated cirrhosis have much better outcomes than those with decompensated disease and may be eligible for interventions to prevent disease progression. METHODS: A cloud-based software solution ("the Liver Toolkit") was developed to access primary care practice software to identify patients at risk of ACLD. Clinical history and laboratory results were extracted to calculate aspartate aminotransferase-to-platelet ratio index and fibrosis 4 scores. Patients identified were recalled for assessment, including Liver Stiffness Measurement (LSM) via transient elastography. Those with an existing diagnosis of cirrhosis were excluded. RESULTS: Existing laboratory results of more than 32,000 adults across nine general practices were assessed to identify 703 patients at increased risk of ACLD (2.2% of the cohort). One hundred seventy-nine patients (26%) were successfully recalled, and 23/179 (13%) were identified to have ACLD (LSM ≥10.0 kPa) (10% found at indeterminate risk [LSM 8.0-9.9 kPa] and 77% low risk of fibrosis [LSM <8.0 kPa]). In most cases, the diagnosis of liver disease was new, with the most common etiology being metabolic dysfunction-associated steatotic liver disease (n=20, 83%). Aspartate aminotransferase-to-platelet ratio index ≥1.0 and fibrosis 4 ≥3.25 had a positive predictive value for detecting ACLD of 19% and 24%, respectively. Patients who did not attend recall had markers of more severe disease with a higher median aspartate aminotransferase-to-platelet ratio index score (0.57 vs. 0.46, p=0.041). CONCLUSIONS: This novel information technology system successfully screened a large primary care cohort using existing laboratory results to identify patients at increased risk ACLD. More than 1 in 5 patients recalled were found to have liver disease requiring specialist follow-up.

A scoping review on occupational science research in European contexts.

BACKGROUND: A survey showed European occupational scientists cover a broad range in occupational science (OS) research, however, no contemporary overviews of European OS research exists, and current research may provide valuable information for OS and occupational therapy. AIM: The aim was to provide an overview of contemporary European OS research. MATERIALS AND METHOD: A scoping review was performed, including studies conducted in Europe and published in the British Journal of Occupational Therapy (BJOT), the Scandinavian Journal of Occupational Therapy (SJOT) or the Journal of Occupational Science (JOS) between 2015 and 2020. The journals were systematically searched, and quality assessment and thematic analysis were undertaken. RESULTS: Findings from 93 articles identified many studies from the Nordic countries. Most studies applied qualitative research methods. Theoretical concepts from OS were used in data generating and discussions. A wide range of demographics, and living conditions were explored. Recent articles took a reflexive stance on the positionality of the researcher/s. CONCLUSIONS: This review highlights the diversity of OS research, suggesting a solid theoretical knowledge base within European OS research. SIGNIFICANCE: The results contribute to further development and maturation of the discipline of OS in Europe and internationally.

Functional reconstitution of an atypical G protein heterotrimer and regulator of G protein signaling protein (RGS1) from Arabidopsis thaliana.

It has long been known that animal heterotrimeric Gαßγ proteins are activated by cell-surface receptors that promote GTP binding to the Gα subunit and dissociation of the heterotrimer. In contrast, the Gα protein from Arabidopsis thaliana (AtGPA1) can activate itself without a receptor or other exchange factor. It is unknown how AtGPA1 is regulated by Gßγ and the RGS (regulator of G protein signaling) protein AtRGS1, which is comprised of an RGS domain fused to a receptor-like domain. To better understand the cycle of G protein activation and inactivation in plants, we purified and reconstituted AtGPA1, full-length AtRGS1, and two putative Gßγ dimers. We show that the Arabidopsis Gα protein binds to its cognate Gßγ dimer directly and in a nucleotide-dependent manner. Although animal Gßγ dimers inhibit GTP binding to the Gα subunit, AtGPA1 retains fast activation in the presence of its cognate Gßγ dimer. We show further that the full-length AtRGS1 protein accelerates GTP hydrolysis and thereby counteracts the fast nucleotide exchange rate of AtGPA1. Finally, we show that AtGPA1 is less stable in complex with GDP than in complex with GTP or the Gßγ dimer. Molecular dynamics simulations and biophysical studies reveal that altered stability is likely due to increased dynamic motion in the N-terminal α-helix and Switch II of AtGPA1. Thus, despite profound differences in the mechanisms of activation, the Arabidopsis G protein is readily inactivated by its cognate RGS protein and forms a stable, GDP-bound, heterotrimeric complex similar to that found in animals.

Decision-making experiences of young adults with long-term conditions.

BACKGROUND: Young adults with long-term conditions can struggle to accept their diagnosis and can become overwhelmed with managing their condition. Suboptimal transfer from paediatric to adult services with a resultant disengagement with the service can result in less involvement in care and decision-making. Shared decision-making can improve involvement in health decisions and increase satisfaction with treatment/therapy and care. OBJECTIVES: An integrative literature review was conducted to explore and understand young adults' experiences of decision-making in health care. DESIGN: An integrative literature review. DATA SOURCES: CINAHL, EMCARE, PsycINFO, HMIC, EMBASE, Web of Science, PubMed, MEDLINE, EBSCOHOST and COCHRANE databases were searched for relevant literature published between January 1999 and January 2020. FINDINGS: Thirteen primary research papers met the inclusion criteria. Four main themes were identified: (1) Information delivery and communication; (2) participation in decision-making; (3) social factors influencing decision-making and (4) emotional impact of decision-making. CONCLUSIONS: Young adults with long-term conditions have specific decision-making needs which can impact their emotional health. Research with a specific focus on young adults' experiences of decision-making in health care is needed.

Promoting Mobility in the Intensive Care Unit: An Educational Interprofessional Initiative.

BACKGROUND: Collaborative interprofessional practice is essential in the critical care arena to promote quality, safety, and positive outcomes for patients. An interprofessional educational activity focusing on early mobilization of intubated intensive care patients was undertaken with senior level nursing students and physical therapy students. Evidence demonstrates that these patients have improved functional outcomes, fewer days requiring mechanical ventilation, fewer days of delirium, and decreased length of stay both within the intensive care unit (ICU) and hospital with early mobilization efforts. METHOD: Students applied knowledge from the literature, clinical guidelines, and evidence-based practices in developing a plan of care to facilitate early mobility of the intensive care intubated patient. A structured debriefing session followed. RESULTS: Students developed a keen appreciation of collaborative, interprofessional, evidence-based care vital to patients who are intubated in the ICU. CONCLUSION: Student engagement in simulated interprofessional teams potentially influences their perceptions and attitudes about its benefits which are necessary for future professional practice. [J Nurs Educ. 2022;61(9):537-541.].

Client resistance to hepatitis C treatment initiation in opioid agonist treatment clinics in Sydney, Australia: A qualitative study.

INTRODUCTION: Opioid agonist treatment (OAT) clinics play a key role in achieving elimination of hepatitis C virus (HCV) globally. Previous research has identified barriers to HCV treatment uptake in OAT clinics; however, most studies were conducted prior to the introduction of direct-acting antiviral treatments (DAA). It remains unclear whether progress has been made in responding to barriers and what challenges persist in this setting. METHODS: Semi-structured in-depth interviews were conducted with staff (n = 20) and clients (n = 15) in two OAT clinics in Sydney, Australia. Interviews were transcribed verbatim and analysed using constant comparative methods. RESULTS: Despite progress in integrating hepatitis C care in the clinics, competing priorities, concerns about side-effects, distrust of staff, health problems and difficulties accessing testing and medication persisted as key reasons why clients had not initiated treatment. Most clients preferred to postpone treatment and focus on other priorities and some highlighted lack of medical evidence for urgent treatment. Pressure on services to achieve elimination targets within set time frames was a primary driver of repeated offers of treatment by staff and the framing of clients' preferences for postponing treatment, as a barrier. DISCUSSION AND CONCLUSION: Current timelines for HCV elimination targets may have galvanised services into action but may have also created tensions at the coalface due to disparities between staff and clients' priorities. The involvement of peer workers and mechanisms to ensure continued follow up with clients about DAA treatments is required. Public health timelines for HCV elimination need to be informed by affected communities' priorities.

Observed communication and associations with satisfaction during traditional and integrative behavioral couple therapy.

To investigate changes in couple communication and potential mechanisms of change during treatment, 134 distressed couples, who were randomly assigned to either traditional or integrative behavioral couple therapy (TBCT; IBCT), were observed in relationship and personal problem discussions prior to and near the end of treatment. Analyses were conducted using the Hierarchical Linear Modeling program. Over the time in therapy, during relationship problem discussions, positivity and problem solving increased while negativity decreased. Compared to IBCT, TBCT couples had the largest gains in positivity and reductions in negativity. During personal problem discussions, negativity decreased, while withdrawal increased and positivity decreased. TBCT couples had larger declines in negativity. In both discussion types, increases in marital satisfaction were associated with increases in positivity and problem solving. Declines in marital satisfaction were associated with increased negativity during relationship problem interactions and increased withdrawal during personal problem interactions. However, no treatment differences in these associations were found. Differences in rule-governed and contingency-shaped behavior change strategies between the two therapies and implications of findings are discussed.

Teaching undergraduate nursing research from an evidence-based practice perspective.

We revised the undergraduate nursing research course using levels of evidence as the template with which to organize the course content and learning activities. A primary purpose in restructuring the course using an evidence-based practice framework was to increase student interest and engagement, and to promote the development of practitioners who will continue to learn about and use research. Learning to systematically locate, evaluate, and use the best available research is the hallmark of evidence-based practice and quality improvement initiatives. Students developed these skills as they sought the answer to a specific clinical problem. One of the most significant outcomes of this approach was students' ability to integrate previously fragmented components of research knowledge and to see the relevance of research evidence for providing excellent patient care.

Demand-withdraw communication in severely distressed, moderately distressed, and nondistressed couples: rigidity and polarity during relationship and personal problem discussions.

This study investigated demand-withdraw communication among 68 severely distressed couples seeking therapy, 66 moderately distressed couples seeking therapy, and 48 nondistressed couples. Self-report and videotaped discussions replicated previous research, demonstrating that greater demand-withdraw during relationship problem discussions was associated with greater distress and that overall, wife-demand/husband-withdraw was greater than husband-demand/wife-withdraw. Results extended the conflict structure view of demand-withdraw by indicating that this gender polarity in demand-withdraw roles varied in strength and direction depending on who chose the topic for discussion, distress level, and marriage length. Further, in videotaped personal problem discussions, typical gender patterns of demand-withdraw were reversed. Across the relationship and personal problem discussions, a pattern of gender polarity emerged when husbands held the burden of changing.

Relative contributions of relationship distress and depression to communication patterns in couples.

Researchers have long been interested in the relationship between marital distress and depression. Empirical findings from investigations into the relative contributions of marital distress and depression to marital communication have been inconsistent, and some communication behaviors, such as the demand/withdraw interaction pattern, have yet to be examined. The ability of depression to predict major types of communication (positive communication, negative communication, problem-solving, and demand/withdraw) was analyzed after controlling for the shared variance between marital distress and depression. Across two studies of couples beginning therapy and one study of couples beginning an enhancement program, results failed to provide support for a unique contribution of depression to couples' communication behaviors.

Quadriphasic waveforms are superior to triphasic waveforms for transthoracic defibrillation in a cardiac arrest swine model with high impedance.

BACKGROUND: We have demonstrated previously that triphasic waveform shocks were superior to biphasic waveform shocks for transthoracic defibrillation. Our purpose was to compare the efficacy and safety of quadriphasic versus triphasic shocks for transthoracic defibrillation in a porcine model. METHODS: Sixteen adult swine (19-25 kg, mean: 21.5 kg) were deeply anesthetized and intubated. To simulate impedance of the human chest, fixed electrical resistors (25 or 50 ohms) was placed in series with the defibrillator and the chest of each pig. After 30 s of electrically induced VF, each pig received transthoracic shocks, using either a truncated exponential triphasic waveform (5 ms positive pulse duration, 5 ms negative pulse duration and 5 ms positive pulse duration, total waveform duration 15 ms) or a quadriphasic waveform (5/5/5/5 ms, total waveform duration 20 ms). Each pig received transthoracic triphasic and quadriphasic shocks at three selected energy levels (50, 100 and 150 J) in random sequence. Four shocks were delivered at each energy level to construct an energy versus % success curve. Success was defined as VF termination at 5 s after shock. The total shocks were divided into three groups based on the delivered energy actually delivered to the animal: <40, 40-65 and >65 J. Delivered energy = (animal impedance/total impedance) times selected energy of the shock. RESULTS: For high-impedance animals (86-102 ohms), quadriphasic waveform shocks achieved significantly higher percent shock success than triphasic shocks for the termination of VF at the energy levels of >65 J actually delivered (quadriphasic 72.7+/-12.2%, triphasic 38.9+/-7.7%, p<0.02). No differences in the shock success between quadriphasic and triphasic waveforms were found for other two energy levels. There were no differences in ventricular tachycardia or asystole after shocks between quadriphasic and triphasic waveforms. CONCLUSION: In this porcine model, 20 ms (5/5/5/5) quadriphasic shocks were superior to 15 ms (5/5/5) triphasic shocks for transthoracic defibrillation in animals with impedances that simulated high impedance in humans.