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The purpose of this study was to compare a wearable microfluidic device and standard absorbent patch in measuring local sweating rate (LSR) and sweat chloride concentration ([Cl-]) in elite basketball players. Participants were 53 male basketball players (25 ± 3 years, 92.2 ± 10.4 kg) in the National Basketball Association's development league. Players were tested during a moderate-intensity, coach-led practice (98 ± 30 min, 21.0 ± 1.2 °C). From the right ventral forearm, sweat was collected using an absorbent patch (3M Tegaderm™ + Pad). Subsequently, LSR and local sweat [Cl-] were determined via gravimetry and ion chromatography. From the left ventral forearm, LSR and local sweat [Cl-] were measured using a wearable microfluidic device and associated smartphone application-based algorithms. Whole-body sweating rate (WBSR) was determined from pre- to postexercise change in body mass corrected for fluid/food intake (ad libitum), urine loss, and estimated respiratory water and metabolic mass loss. The WBSR values predicted by the algorithms in the smartphone application were also recorded. There were no differences between the absorbent patch and microfluidic patch for LSR (1.25 ± 0.91 mg·cm-2·min-1 vs. 1.14 ±0.78 mg·cm-2·min-1, p = .34) or local sweat [Cl-] (30.6 ± 17.3 mmol/L vs. 29.6 ± 19.4 mmol/L, p = .55). There was no difference between measured and predicted WBSR (0.97 ± 0.41 L/hr vs. 0.89 ± 0.35 L/hr, p = .22; 95% limits of agreement = 0.61 L/hr). The wearable microfluidic device provides similar LSR, local sweat [Cl-], and WBSR results compared with standard field-based methods in elite male basketball players during moderate-intensity practices.

Extended and immersive gamified pharmacy simulation has been demonstrated to provide transformative learning in pharmacy education, preparing graduates for real-world practice. An international consortium of universities has implemented local adaptations of the Pharmacy Game into their curricula. From early 2020, pharmacy academics modified the delivery of gamified simulation in response to the COVID-19 pandemic, while still aiming to deliver the important learning outcomes of enhanced communication, collaboration, confidence and competence. Australian universities went into full lockdown from March 2020, and the critical gamified simulation at Griffith University was delivered entirely virtually in 2020. An array of synchronous and asynchronous approaches and software platforms was employed, including Microsoft Teams, Forms and Stream plus the online interview platform Big Interview. These allowed for the simulation activities, including dispensing, counselling and clinical cases, to be conducted by students online. In 2021, Griffith University conducted hybrid delivery of its Pharmacy Game, balancing student participation both in person and online. Microsoft Power Apps was added to the hosting platform to enhance the simulation interface, and Power Virtual Agent artificial intelligence chatbots, with natural language processing, were used to enable asynchronous clinical interaction. The combination of learning technologies provided the means to deliver successful gamified simulation in the virtual and hybrid environments while still achieving outstanding learning outcomes from the capstone activity. This paper details the technologies used to virtualize the Australian Pharmacy Game and the analytics available to educators to assess student participation, engagement and performance.

Sports nutrition is a relatively new discipline; with ~100 published papers/year in the 1990s to ~3,500+ papers/year today. Historically, sports nutrition research was primarily initiated by university-based exercise physiologists who developed new methodologies that could be impacted by nutrition interventions (e.g., carbohydrate/fat oxidation by whole body calorimetry and muscle glycogen by muscle biopsies). Application of these methods in seminal studies helped develop current sports nutrition guidelines as compiled in several expert consensus statements. Despite this wealth of knowledge, a limitation of the current evidence is the lack of appropriate intervention studies (e.g., randomized controlled clinical trials) in elite athlete populations that are ecologically valid (e.g., in real-life training and competition settings). Over the last decade, there has been an explosion of sports science technologies, methodologies, and innovations. Some of these recent advances are field-based, thus, providing the opportunity to accelerate the application of ecologically valid personalized sports nutrition interventions. Conversely, the acceleration of novel technologies and commercial solutions, especially in the field of biotechnology and software/app development, has far outstripped the scientific communities' ability to validate the effectiveness and utility of the vast majority of these new commercial technologies. This mini-review will highlight historical and present innovations with particular focus on technological innovations in sports nutrition that are expected to advance the field into the future. Indeed, the development and sharing of more "big data," integrating field-based measurements, resulting in more ecologically valid evidence for efficacy and personalized prescriptions, are all future key opportunities to further advance the field of sports nutrition., Competing Interests: IR and MK are employees of the Gatorade Sports Science Institute, a division of PepsiCo, Inc. KJ, TS, and YP, received speaking honoraria, for the GSSI ECSS 2021 pre-conference symposium which inspired this article. YP is the founding member of the Sub2 project (www.sub2hrs.com). The Sub2 project is affiliated to a non-trading company (Athlome Limited, UK) that is minor (<1.1%) shareholder of Maurten AB, Gothenburg, Sweden., (Copyright © 2022 Jonvik, King, Rollo, Stellingwerff and Pitsiladis.)

Adaptive mutations can cause drug resistance in cancers and pathogens, and increase the tolerance of agricultural pests and diseases to chemical treatment. When and how adaptive mutations form is often hard to discern, but we have shown that adaptive copy number amplification of the copper resistance gene CUP1 occurs in response to environmental copper due to CUP1 transcriptional activation. Here we dissect the mechanism by which CUP1 transcription in budding yeast stimulates copy number variation (CNV). We show that transcriptionally stimulated CNV requires TREX-2 and Mediator, such that cells lacking TREX-2 or Mediator respond normally to copper but cannot acquire increased resistance. Mediator and TREX-2 can cause replication stress by tethering transcribed loci to nuclear pores, a process known as gene gating, and transcription at the CUP1 locus causes a TREX-2-dependent accumulation of replication forks indicative of replication fork stalling. TREX-2-dependent CUP1 gene amplification occurs by a Rad52 and Rad51-mediated homologous recombination mechanism that is enhanced by histone H3K56 acetylation and repressed by Pol32 and Pif1. CUP1 amplification is also critically dependent on late-firing replication origins present in the CUP1 repeats, and mutations that remove or inactivate these origins strongly suppress the acquisition of copper resistance. We propose that replicative stress imposed by nuclear pore association causes replication bubbles from these origins to collapse soon after activation, leaving a tract of H3K56-acetylated chromatin that promotes secondary recombination events during elongation after replication fork re-start events. The capacity for inefficient replication origins to promote copy number variation renders certain genomic regions more fragile than others, and therefore more likely to undergo adaptive evolution through de novo gene amplification., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Potential cochlear implant (CI) candidates arrive to the clinic with a variety of hearing loss configurations, hearing aid history, and aided capabilities. CI candidacy is primarily determined based on aided speech recognition capability, which relies on benefit derived from use of hearing aids. Therefore, contemporary evaluation for CI candidacy should incorporate a battery of testing to determine abilities and limitations and must be predicated on appropriate verification of the hearing aid fitting. However, recent reports, including a retrospective chart review of patients presenting to Cleveland Clinic for CI evaluation, suggest that a significant subset of patients may be using inappropriately fit or programmed amplification. Thus, a combination of simulated real-ear measurements and aided speech recognition testing is essential for fully assessing the effect of amplification and ultimately determination of CI candidacy. Furthermore, waiting to incorporate these tools until CI candidacy is suspected may delay timely identification of problems or need to change technology. Utilization of evidence-based decision drivers ultimately leads clinicians to timely patient-specific interventions which may include surgical intervention or other amplification options. As audiology moves into a healthcare era in which payers consider the benefit of our services to overall health and well-being, demonstrating timely, optimal outcomes using thorough, multifactorial evaluation is essential., Competing Interests: Conflict of Interest S.A.S. is a consultant for Oticon Medical and Cochlear., (Thieme. All rights reserved.)

The purpose of the study was to determine if repeated exertional heat injuries (EHIs) worsen the inflammatory response. We assessed the impact of a single EHI bout (EHI0) or two separate EHI episodes separated by 1 (EHI1), 3 (EHI3), and 7 (EHI7) days in male C57BL/6J mice ( n = 236). To induce EHI, mice underwent a forced running protocol until loss of consciousness or core temperature reached ≥ 42.7°C. Blood and tissue samples were obtained 30 min, 3 h, 1 day, or 7 days after the EHI. We observed that mice undergoing repeated EHI (EHI1, EHI3, and EHI7) had longer running distances before collapse (∼528 m), tolerated higher core temperatures (∼0.18°C higher) before collapse, and had higher minimum core temperature (indicative of injury severity) during recovery relative to EHI0 group (∼2.18°C higher; all P < 0.05). Heat resilience was most pronounced when latency was shortest between EHI episodes (i.e., thermal load and running duration highest in EHI1), suggesting the response diminishes with longer recoveries between EHI events. Furthermore, mice experiencing a second EHI exhibited increased serum and liver HSP70, and lower corticosterone, FABP2, MIP-1β, MIP-2, and IP-10 relative to mice experiencing a single EHI typically at 30 min to 3 h after EHI. Our findings indicate that an EHI event may initiate some adaptive processes that provide acute heat resilience to subsequent EHI conditions. NEW & NOTEWORTHY Mice undergoing repeated exertional heat injuries, within 1 wk of an initial heat injury, appear to have some protective adaptations. During the second exertional heat injury, mice were able to run longer and sustain higher body temperatures before collapse. Despite this, the mice undergoing a second exertional heat injury were more resilient to the heat as evidenced by attenuated minimum body temperature, higher HPS70 (serum and liver), lower corticosterone, and lower FABP2.

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 °C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 °C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 °C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or naïve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

Intestinal barrier integrity and function are compromised during exertional heat stress (EHS) potentially leading to consequences that range from minor gastrointestinal (GI) disturbances to fatal outcomes in exertional heat stroke or septic shock. This mini-review provides a concise discussion of nutritional interventions that may protect against intestinal permeability during EHS and suggests physiological mechanisms responsible for this protection. Although diverse nutritional interventions have been suggested to be protective against EHS-induced GI permeability, the ingestion of certain amino acids, carbohydrates, and fluid per se is potentially effective strategy, whereas evidence for various polyphenols and pre/probiotics is developing. Plausible physiological mechanisms of protection include increased blood flow, epithelial cell proliferation, upregulation of intracellular heat shock proteins, modulation of inflammatory signaling, alteration of the GI microbiota, and increased expression of tight junction (TJ) proteins. Further clinical research is needed to propose specific nutritional candidates and recommendations for their application to prevent intestinal barrier disruption and elucidate mechanisms during EHS.

Pseudomonas aeruginosa is an opportunistic pathogen causing life-threatening infections. Previously, we showed that elevated calcium (Ca 2+ ) levels increase the production of virulence factors in P. aeruginosa In an effort to characterize the Ca 2+ regulatory network, we identified a Ca2+ - r egulated β- p ropeller protein, CarP, and showed that expression of the encoding gene is controlled by the Ca 2+ -regulated two-component system CarSR. Here, by using a Galleria melonella model, we showed that CarP plays a role in regulating P. aeruginosa virulence. By using transcriptome sequencing (RNA-Seq), reverse transcription (RT)-PCR, quantitative RT-PCR (RT-qPCR), and promoter fusions, we determined that carP is transcribed into at least two transcripts and regulated by several bacterial and host factors. The transcription of carP is elevated in response to Ca 2+ in P. aeruginosa cystic fibrosis isolates and PAO1 laboratory strain. Elevated Fe 2+ also induces carP The simultaneous addition of Ca 2+ and Fe 2+ increased the carP promoter activity synergistically, which requires the presence of CarR. In silico analysis of the intergenic sequence upstream of carP predicted recognition sites of RhlR/LasR, OxyR, and LexA, suggesting regulation by quorum sensing (QS) and oxidative stress. In agreement, the carP promoter was activated in response to stationary-phase PAO1 supernatant and required the presence of elevated Ca 2+ and CarR but remained silent in the triple mutant lacking rhlI, lasI , and pqsA synthases. We also showed that carP transcription is regulated by oxidative stress and that CarP contributes to P. aeruginosa Ca 2+ -dependent H 2 O 2 tolerance. The multifactorial regulation of carP suggests that CarP plays an important role in P. aeruginosa adaptations to host environments. IMPORTANCE P. aeruginosa is a human pathogen causing life-threatening infections. It is particularly notorious for its ability to adapt to diverse environments within the host. Understanding the signals and the signaling pathways enabling P. aeruginosa adaptation is imperative for developing effective therapies to treat infections caused by this organism. One host signal of particular importance is calcium. Previously, we identified a component of the P. aeruginosa calcium-signaling network, CarP, whose expression is induced by elevated levels of calcium. Here, we show that carP plays an important role in P. aeruginosa virulence and is upregulated in P. aeruginosa strains isolated from sputa of patients with cystic fibrosis. We also identified several bacterial and host factors that regulate the transcription of carP Such multifactorial regulation highlights the interconnectedness between regulatory circuits and, together with the pleotropic effect of CarP on virulence, suggests the importance of this protein in P. aeruginosa adaptations to the host., (Copyright © 2021 American Society for Microbiology.)

Background: Worldwide, pharmacy practice is changing to include new roles and responsibilities. Laws enabling the implementation of assisted dying are expanding in international jurisdictions. Pharmacy practice in assisted dying is subsequently expanding. However, studies of how pharmacists experience their practice when engaged in assisted dying are absent. To progress research into the lived experiences of pharmacists practicing in assisted dying, the development of an inquiry framework to guide such research is the first step., Objective: The objective was to develop a theoretical framework of inquiry for use in subsequent continuing research which may explore the actual experience of pharmacy practice in assisted dying., Methods: Perspectives were gathered from expert and senior pharmacists who were anticipating the imminent implementation of assisted dying practice. Analysis focused on understanding what aspects of practice experience were important to them. Interview-conversations centred on the question: If you had the chance to talk to experienced pharmacist practitioners who have been involved in the practice of assisted dying, what aspects regarding their experiences, would you like to know about? A conventional approach to qualitative content analysis was utilized to analyze the data., Results: Findings summarized questions posed by pharmacists contemplating the implementation of assisted dying practice. These perspectives formed the foundation of a theoretical inquiry framework constituted by 8 inter-related dimensional range-continuums. Each range-continuum, designed to explore the lived experiences of pharmacists in practice, is defined. Examples of how the inquiry dimensions will be used to inform future exploratory research are offered within the framework., Conclusions: The theoretical inquiry framework will be used to develop knowledge for pharmacists contemplating participation (or not) in assisted dying practice. It is timely to progress research that reveals the informed experiences of pharmacists that are actually practicing in this area. The framework may be adapted for researching pharmacists' experience in other practice areas and contexts., (Crown Copyright © 2020. Published by Elsevier Inc. All rights reserved.)

Background: Studies have explored community pharmacy ethical dilemmas; however, limited research exists on hospital pharmacy ethical issues and pharmacists' ethical decision-making processes. Research exploring this is timely, considering developments in hospital pharmacy practices, new hospital pharmacist roles, and evolving responsibilities., Aim/objectives: To explore hospital pharmacists' ethical decision-making and processes for managing ethical challenges in the context of evolving Australian hospital pharmacy practices., Methods: Face-to-face semi-structured interviews with 20 purposively-selected hospital pharmacists from four Queensland Health hospitals. An interview guide with 11 open-ended questions and prompts was developed, validated, and trialed. Pharmacists who consented received the guide prior to interviews. Interviews were audio recorded, transcribed verbatim, and compared with field notes. Transcribed data were imported into NVivo 12 to facilitate coding and thematic analysis., Results: Participants were interviewed January to April 2019; median interview duration was 17.45 min. Data saturation was reached. Participants' experiences ranged from junior level pharmacists to senior management positions, in clinical and non-clinical roles. Emerging themes were: 1) influences on the development of ethical decision-making skills, 2) ethical decision-making is an integral part of the hospital pharmacist's role, and 3) institutional requirements and settings impact on ethical exposure. A wide range of contemporary ethical issues unique to hospital pharmacy practice, mostly involving complex medication management safety, supply, and cost scenarios, were identified. Junior pharmacists indicated they would benefit from additional training, mentorship, and availability of hospital-specific targeted ethics resources., Conclusion: The findings highlighted that hospital pharmacists are regularly faced with ethical issues unique to the hospital pharmacy practice context. Application of sound and structured ethical reasoning and decision-making is, therefore, required in this setting. Participants identified many interrelated factors that impacted their ethical reasoning and behaviour. This study identified gaps that, once addressed, will better support ethical reasoning in hospital pharmacy settings., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Pseudomonas aeruginosa infects patients with cystic fibrosis, burns, wounds and implants. Previously, our group showed that elevated Ca 2+ positively regulates the production of several virulence factors in P. aeruginosa , such as biofilm formation, production of pyocyanin and secreted proteases. We have identified a Ca 2+ -regulated β-propeller putative phytase, CarP, which is required for Ca 2+ tolerance, regulation of the intracellular Ca 2+ levels, and plays a role in Ca 2+ regulation of P. aeruginosa virulence. Here, we studied the conservation of carP sequence and its occurrence in diverse phylogenetic groups of bacteria. In silico analysis revealed that carP and its two paralogues PA2017 and PA0319 are primarily present in P. aeruginosa and belong to the core genome of the species. We identified 155 single nucleotide alterations within carP , 42 of which lead to missense mutations with only three that affected the predicted 3D structure of the protein. PCR analyses with carP -specific primers detected P. aeruginosa specifically in 70 clinical and environmental samples. Sequence comparison demonstrated that carP is overall highly conserved in P. aeruginosa isolated from diverse environments. Such evolutionary preservation of carP illustrates its importance for P. aeruginosa adaptations to diverse environments and demonstrates its potential as a biomarker.

Objectives: Reclassification of medicines from prescription to non-prescription increases timely access to treatment, promotes self-management of minor ailments and relieves healthcare system burden. Previous research identified that Australia lagged behind the United Kingdom and New Zealand in medicines reclassification. This study aimed to identify Australian pharmacists' opinions on the current state of medicines reclassification; the prescription medicines consumers requested without prescription; the medicines pharmacists believed should and should not be considered for reclassification; and perceived barriers to reclassification., Methods: A 2016 national online survey that sought pharmacists' opinions on the state of reclassification, perceived barriers to reclassification and readiness of the profession for further reclassification. Pharmacists' comments were invited through open-ended questions., Key Findings: Two hundred and thirty-five valid surveys were completed. Respondents practised in community, hospital, consultant and academic contexts, and the majority were female (58.7%, n = 138). More than two thirds (70.66%, n = 166) of pharmacists reported receiving daily or weekly requests for non-prescription access to prescription medicines. The majority of pharmacists (71.7%) agreed that the Australian pharmacy profession is ready for further medicines reclassification, guided by patient safety, harm minimisation and medication continuance. The most prominent barrier to further reclassification was opposition from other healthcare professionals., Conclusions: Australian pharmacists believe that their profession has the capacity to safely and effectively manage a wider range of non-prescription medicines through increased reclassification in the contexts of patient safety and risk mitigation. This study has contributed to the global conversation on non-prescription medicines access, providing momentum for practice and policy change., (© 2020 Royal Pharmaceutical Society.)

Background: The aim of this study was to characterize the time-resolved progression of clinical laboratory disturbances days-following an exertional heat stroke (EHS). Currently, normalization of organ injury clinical biomarker values is the primary indicator of EHS recovery. However, an archetypical biochemical recovery profile following EHS has not been established., Methods: We performed a retrospective analysis of EHS patient records in US military personnel from 2008-2014 using the Military Health System Data Repository (MDR). We focused on commonly reported clinical laboratory analytes measured on the day of injury and all proceeding follow-up visits., Results: Over the prescribed period, there were 2,529 EHS episodes treated at 250 unique treatment locations. Laboratory results, including a standardized set of blood, serum and urine assays, were analyzed from 0-340 days following the initial injury. Indicators of acute kidney injury, including serum electrolyte disturbances and abnormal urinalysis findings, were most prevalent on the day of the injury but normalized within 24-48hours (creatinine, blood urea nitrogen, and blood and protein in urine). Muscle damage and liver function-associated markers peaked 0-4 days after injury and persisted outside their respective reference ranges for 2-16 days (alanine aminotransferase, aspartate aminotransferase, creatine phosphokinase, myoglobin, prothrombin time)., Conclusion: Biochemical recovery from EHS spans a 16-day time course, and markers of end-organ damage exhibit distinct patterns over this period. This analysis underscores the prognostic value of each clinical laboratory analyte and will assist in evaluating EHS patient presentation, injury severity and physiological recovery., Competing Interests: The authors have declared that no competing interests exist.

Background: The scope and roles of pharmacists worldwide are undergoing dramatic change. Patient-focused care aimed at caring for people that seek medical assistance in dying is among the newest roles. While pharmacists have been involved in medically assisted dying in some international jurisdictions for over two decades, little is known about their actual lived experiences., Objective: To map the literature concerning pharmacy practice in the assisted dying domain to clarify apparent research gaps., Methods: A mapping review was preformed following a systematic search of Medline, CINAHL and IPA to locate academic papers and reports relating to pharmacists' involvement in assisted dying published between 1990 and 2019. Searches included articles in English, French, and Dutch. References and citations of articles were searched to identify additional articles., Results: A total of 43 articles were selected, including commentaries (n = 26), reports (n = 2), a scoping literature review (n = 1), and empirical studies (n = 14). Most commentaries centered on pharmacists' roles, ethico-legal and moral challenges, and educational concerns in relation to participation. Of the 14 empirical studies, 12 studies were designed around surveys that focused on pharmacists' attitudes, and opinions concerning assisted dying. Other methodologies included thematic analysis of moral dilemmas, experimental design identifying attitudes to sedation at end of life, and analysis of documents such as guidelines, position statements, and standards of practice. Two studies utilized a qualitative research approach. A significant gap was found with respect to research exploring the actual experience of pharmacists' practice in medically assisted dying., Conclusion: There is an absence of studies exploring pharmacists' actual experiences in assisted dying practice. Research involving pharmacists that participate in legally sanctioned assisted dying will facilitate a meaningful understanding of the lived experience of pharmacy practice in this domain., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Calcium (Ca 2+ ) is a universal signaling ion, whose major informational role shaped the evolution of signaling pathways, enabling cellular communications and responsiveness to both the intracellular and extracellular environments. Elaborate Ca 2+ regulatory networks have been well characterized in eukaryotic cells, where Ca 2+ regulates a number of essential cellular processes, ranging from cell division, transport and motility, to apoptosis and pathogenesis. However, in bacteria, the knowledge on Ca 2+ signaling is still fragmentary. This is complicated by the large variability of environments that bacteria inhabit with diverse levels of Ca 2+ . Yet another complication arises when bacterial pathogens invade a host and become exposed to different levels of Ca 2+ that (1) are tightly regulated by the host, (2) control host defenses including immune responses to bacterial infections, and (3) become impaired during diseases. The invading pathogens evolved to recognize and respond to the host Ca 2+ , triggering the molecular mechanisms of adhesion, biofilm formation, host cellular damage, and host-defense resistance, processes enabling the development of persistent infections. In this review, we discuss: (1) Ca 2+ as a determinant of a host environment for invading bacterial pathogens, (2) the role of Ca 2+ in regulating main events of host colonization and bacterial virulence, and (3) the molecular mechanisms of Ca 2+ signaling in bacterial pathogens.

Extrachromosomal circular DNA (eccDNA) facilitates adaptive evolution by allowing rapid and extensive gene copy number variation and is implicated in the pathology of cancer and ageing. Here, we demonstrate that yeast aged under environmental copper accumulate high levels of eccDNA containing the copper-resistance gene CUP1. Transcription of the tandemly repeated CUP1 gene causes CUP1 eccDNA accumulation, which occurs in the absence of phenotypic selection. We have developed a sensitive and quantitative eccDNA sequencing pipeline that reveals CUP1 eccDNA accumulation on copper exposure to be exquisitely site specific, with no other detectable changes across the eccDNA complement. eccDNA forms de novo from the CUP1 locus through processing of DNA double-strand breaks (DSBs) by Sae2, Mre11 and Mus81, and genome-wide analyses show that other protein coding eccDNA species in aged yeast share a similar biogenesis pathway. Although abundant, we find that CUP1 eccDNA does not replicate efficiently, and high-copy numbers in aged cells arise through frequent formation events combined with asymmetric DNA segregation. The transcriptional stimulation of CUP1 eccDNA formation shows that age-linked genetic change varies with transcription pattern, resulting in gene copy number profiles tailored by environment., Competing Interests: The authors have declared that no competing interests exist.

Objectives: To investigate the use, understanding, trust and influence of the internet and other sources of health information used by the next of kin (NOK) of patients admitted to the intensive care unit (ICU)., Design: Multicentre structured survey., Setting: The ICUs of 13 public and private Australian hospitals., Participants: NOK who self-identified as the primary surrogate decision maker for a patient admitted to the ICU., Main Outcome Measures: The frequency, understanding, trust and influence of online sources of health information, and the quality of health websites visited using the Health on the Net Foundation Code of Conduct (HONcode) for medical and health websites., Results: There were 473 survey responses. The median ICU admission days and number of ICU visits by the NOK at the time of completing the survey was 3 (IQR, 2-6 days) and 4 (IQR, 2-7), respectively. The most commonly reported sources of health information used very frequently were the ICU nurse (55.6%), ICU doctor (38.7%), family (23.3%), hospital doctor (21.4%), and the internet (11.3%). Compared with the 243 NOK (51.6%) not using the internet, NOK using the internet were less likely to report complete understanding (odds ratio [OR], 0.57; 95% CI, 0.38-0.88), trust (OR, 0.34; 95% CI, 0.19-0.59), or influence (OR, 0.58; 95% CI, 0.38-0.88) associated with the ICU doctor. Overall, the quality of the 40 different reported websites accessed was moderately high., Conclusions: A substantial proportion of ICU NOK report using the internet as a source of health information. Internet use is associated with lower reported understanding, trust and influence of the ICU doctor.

Introduction: Precipitating factors that contribute to the severity of exertional heat stroke (EHS) are unclear. The purpose of this study was to determine the effect of prior illness (PI) on EHS severity., Methods: We performed a retrospective clinical record review of 179 documented cases of EHS at the Marine Corps Base in Quantico, Virginia., Results: Approximately 30% of EHS cases had a medically documented PI. Anthropometrics (height, weight, body mass index) and commonly associated risk factors for EHS (age, number of days in training, wet bulb globe temperature, sleep patterns) did not differ between PI and no illness (NI) groups. PI patients presented with higher maximal rectal core temperatures (40.6 ± 1.0°C vs. 40.3 ± 1.2°C; P = 0.0419), and elevated pulse rates (118.1 ± 16.7 bpm vs. 110.5 ± 24.2 bpm; P = 0.0397). At the point of care, biomarker values were similar between PI and NI groups, with the exception of a trend toward elevated monocytes in those with PI (7.9 ± 2.9% vs 6.7± 2.7%; P = 0.0521). Rate and duration of cooling were similar between PI and NI patients., Conclusion: This study indicates that PI has a minimal effect on the patient presentation, severity and treatment outcome of EHS. The results of this study have important implications for military, civilian, and occupational populations who are at risk for EHS., Competing Interests: The authors have declared that no competing interests exist.

Background Ethical reasoning informs decision making and professional judgement, is guided by codes of ethics and conduct, and requires navigation through a regulatory framework. Ethical reasoning should evolve throughout the pharmacy internship year and prepare interns for independent practice. Objective To explore the ethical reasoning and processes of Australian pharmacists and pharmacy interns. Setting Queensland community pharmacists and interns. Method A survey to determine use of resources to guide ethical decisions, management of ethical dilemmas, and exposure to potential practice privacy breaches. Participants were recruited at pharmacy intern training events, a pharmacist education session and through telephone contact of randomised community pharmacies. Main outcome measure Comparison between pharmacist and intern responses using 5-point Likert scales, listings and prioritising. Results In total 218 completed surveys were analysed: 121 pharmacy interns and 97 pharmacists. The Code of Ethics was identified as the resource most frequently consulted when faced with ethical dilemmas. Interns were more likely to consult legislation and regulatory authorities whereas pharmacists with colleagues. Responses to ethical vignette scenarios and exposure to privacy breaches varied between interns and pharmacists, with some scenarios revealing significant differences. Most participants had been exposed to a variety of potential privacy breaches in practice. Conclusion Interns focussed on legislation and guidelines when presented with hypothetical ethical dilemmas. In contrast to this positivist approach, pharmacists reported using a social constructionist approach with peers as a reference. Pharmacists avoided ethical scenario options that required complex management. Interns reported more exposure to potential practice privacy breaches.

MicroRNA-1 (miRNA-1) has been long viewed as a muscle-specific miRNA and plays a critical role in myocardium and cardiomyocytes by controlling myocyte growth and rhythm. We identified that miRNA-1 is expressed in cardiac fibroblasts, which are one of the major non-muscle cell types in myocardium and are responsible for cardiac fibrosis in pathological conditions. In this study, we aimed to investigate the effect and mechanism of action of miRNA-1 on cardiac fibroblast proliferation. Subcutaneous angiotensin II (Ang II) infusion via osmotic minipumps for 4 weeks was used to induce myocardial interstitial fibrosis in male Sprague-Dawley rats. MiRNA-1 expression was significantly down-regulated by 68% in freshly isolated ventricular fibroblasts from Ang II-infused rats than that from control rats. Similar results were obtained in adult rat ventricular fibroblasts that were stimulated in culture by Ang II or TGFβ for 48 h. Functionally, overexpression of miRNA-1 inhibited fibroblast proliferation, whereas knockdown of endogenous miRNA-1 increased fibroblast proliferation. We then identified and validated cyclin D2 and cyclin-dependent kinase 6 (CDK6) as direct targets of miRNA-1 in cardiac fibroblasts using biochemical assays. Moreover, we showed that the inhibitory effects of miRNA-1 on cardiac fibroblast proliferation can be blunted by overexpression of its target, cyclin D2. In conclusion, our findings demonstrate miRNA-1 expression and regulation in adult ventricular fibroblasts, where it acts as a novel negative regulator of adult cardiac fibroblast proliferation that is at least partially mediated by direct targeting of two cell cycle regulators. Our results expand the understanding of the regulatory roles of miRNA-1 in cardiac cells (i.e., from myocytes to a major non-muscle cells in the heart).

Objectives: Availability of medicines without prescription can increase consumers' timely access to treatment and promote self-management of minor ailments and adherence to long-term medications. Globally, access to relevant medicines has improved through increased reclassification of medicines from prescription to non-prescription availability. However, Australian reclassification lags behind countries with comparable health systems, and the factors influencing this are poorly understood., Methods: Semi-structured interviews were conducted during May 2015 to explore the perspectives of Australian pharmacists and support staff on future reclassification. Interview responses were transcribed verbatim, and the data were analysed thematically, primarily informed by the general inductive approach., Results: Participants identified a broad range of medicines as candidates for future reclassification by applying risk versus benefit judgements, assessing any medicines with potential for misuse and hazardous medicines as unsuitable. Key drivers for change in classification were underpinned by participants' desire to support consumers' management of minor ailments and adherence for those on long-term therapy. Barriers to reclassification were identified by pharmacy staff as internal, negatively impacting pharmacists' readiness for reclassification and external, negatively impacting the overall progress of change., Conclusions: While the research provided valuable insights to inform the ongoing discussion on future reclassification, a larger, more representative sample is needed to confirm these findings.

The feasibility of an individualised carer support service delivered in community pharmacies was assessed from the perspective of carer participants using a pre-post questionnaire and semistructured interviews. Eligible pharmacies were required to offer a medication management service relevant to carers and have a semiprivate space for conversations. Carers were required to self-identify as an unpaid support person for someone with a chronic condition or disability. Between September 2016 and March 2017, staff from 11 community pharmacies in South-East Queensland, Australia were trained, and provided with ongoing mentoring from a pharmacist and carer to support service implementation. Identification of carers and support to achieve a personal and care-giving goal were key features of the service. Questionnaires included the EQ-5D-3L, the Bakas Caregiving Outcomes Scale, and questions relating to goal achievement, carer roles, and responsibilities. Seven follow-up carer interviews were undertaken between March and May 2017 and analysed thematically. Pre-post questionnaires were available for 17 carers (one withdrew, two incomplete). Of the 29 goals set, 10 were achieved and 14 partially achieved. EQ-5D-3L scores were unchanged, while 7 of the 15 items comprising the Bakas score improved (p < 0.05). Carer service evaluation was generally favourable, and these two main interview themes were the impact of caring and pharmacy experience. The impact of caring, while variable, was significant. Pharmacy experiences were mostly positive and the opportunity for carers to further engage with pharmacy staff was appreciated. The service was feasible and initial reported benefits to carers may support further research potentially in terms of a larger controlled trial., (© 2018 John Wiley & Sons Ltd.)

Objectives: Unpaid carers have many and varied responsibilities in society, which can include medication management for the person they support. However, the potential for Australian community pharmacies to better assist carers is relatively unexplored. This mixed-methods study investigated the acceptability of a local carer support service by trained community pharmacy staff, including issues regarding the implementation and impact of this service., Methods: Staff from 11 community pharmacies in South East Queensland, Australia, were trained to deliver a six-step carer support service between September 2016 and March 2017. Pharmacies were supported by a carer and pharmacist mentor pair and asked to recruit up to six carers each. Evaluations of staff training were descriptively analysed. Semi-structured interviews were undertaken with pharmacy staff, and interview transcripts were analysed thematically., Key Findings: Staff training evaluations were positive; participants acquired new information about carers and rated the service highly in terms of its importance within the pharmacy setting. Feedback was obtained on how to improve the training, such as further opportunities for role-play. Seven staff members were interviewed, and data analysis revealed two main themes: (1) implementation of the carer support service and (2) perceived impact on pharmacy staff. Positive attitudes towards recognising and supporting carers, and training and mentoring were identified with community pharmacies viewed as a suitable place for delivering this new service. New insights into the impact of caring were widely reported, which staff had not appreciated from previous carer interactions. Structural issues, including space and time pressures, and a lack of awareness about the types of support currently available to carers were emphasised., Conclusion: Pharmacy staff are well positioned to support carers. Engaging carers in conversation to better understand their needs is a small step with potential for big gains, including a more empathetic understanding of their individual circumstances and overall well-being., (© 2018 Royal Pharmaceutical Society.)

Transcription of protein coding genes is accompanied by recruitment of COMPASS to promoter-proximal chromatin, which methylates histone H3 lysine 4 (H3K4) to form H3K4me1, H3K4me2 and H3K4me3. Here, we determine the importance of COMPASS in maintaining gene expression across lifespan in budding yeast. We find that COMPASS mutations reduce replicative lifespan and cause expression defects in almost 500 genes. Although H3K4 methylation is reported to act primarily in gene repression, particularly in yeast, repressive functions are progressively lost with age while hundreds of genes become dependent on H3K4me3 for full expression. Basal and inducible expression of these genes is also impaired in young cells lacking COMPASS components Swd1 or Spp1. Gene induction during ageing is associated with increasing promoter H3K4me3, but H3K4me3 also accumulates in non-promoter regions and the ribosomal DNA. Our results provide clear evidence that H3K4me3 is required to maintain normal expression of many genes across organismal lifespan., Competing Interests: CC, MD, CK, QG, DH, MK, LF, JH No competing interests declared, (© 2018, Cruz et al.)

With increasing participation of females in endurance athletics and active military service, it is important to determine if there are inherent sex-dependent susceptibilities to exertional heat injury or heat stroke. In this study we compared responses of male and female adult mice to exertional heat stroke (EHS). All mice were instrumented for telemetry core temperature measurements and were exercise-trained for 3 wk before EHS. During EHS, environmental temperature was 37.5°C (35% RH) while the mice ran on a forced running wheel, using incremental increases in speed. The symptom-limited endpoint was loss of consciousness, occurring at ~42.2°C core temperature. Females ran greater distances (623 vs. 346 m, P < 0.0001), reached faster running speeds (7.2 vs. 5.1 m/min, P < 0.0001), exercised for longer times (177 vs. 124 min, P < 0.0001), and were exposed to greater internal heat loads (240 vs.160°C·min; P < 0.0001). Minimum Tc during hypothermic recovery was ~32.0°C in both sexes. Females lost 9.2% body weight vs. 7.5% in males ( P < 0.001). Females demonstrated higher circulating corticosterone (286 vs 183 ng/ml, P = 0.001, at 3 h), but most plasma cytokines were not different. A component of performance in females could be attributed to greater body surface area/mass and greater external power performance. However, there were significant and independent effects of sex alone and a crossed effect of "sex × power" on performance. These results demonstrate that female mice have greater resistance to EHS during exercise in hyperthermia and that these effects cannot be attributed solely to body size. NEW & NOTEWORTHY Female mice are surprisingly more resistant to exertional heat stroke than male mice. They run faster and longer and can withstand greater internal heat loads. These changes cannot be fully accounted for by increased body surface/mass ratio in females or on differences in aerobic performance. Although the stress-immune response in males and females was similar, females exhibited markedly higher plasma corticosteroid levels, which were sustained over 14 days of recovery.

Background: The limited neurobiological understanding of posttraumatic stress disorder (PTSD) has been partially attributed to the need for improved animal models. Stress-enhanced fear learning (SEFL) in rodents recapitulates many PTSD-associated behaviors, including stress-susceptible and stress-resilient subgroups in outbred rats. Identification of subgroups requires additional behavioral phenotyping, a confound to mechanistic studies., Methods: We employed a SEFL paradigm in inbred male and female C57BL/6 mice that combines acute stress with fear conditioning to precipitate traumatic-like memories. Extinction and long-term retention of extinction were examined after SEFL. Further characterization of SEFL effects on male mice was performed with additional behavioral tests, determination of regional activation by Fos immunofluorescence, and RNA sequencing of the basolateral amygdala., Results: Stressed animals displayed persistently elevated freezing during extinction. While more uniform in females, SEFL produced male subgroups with differential susceptibility that were identified without posttraining phenotyping. Additional phenotyping of male mice revealed PTSD-associated behaviors, including extinction-resistant fear memory, hyperarousal, generalization, and dysregulated corticosterone in stress-susceptible male mice. Altered Fos activation was also seen in the infralimbic cortex and basolateral amygdala of stress-susceptible male mice after remote memory retrieval. Key behavioral outcomes, including susceptibility, were replicated by two independent laboratories. RNA sequencing of the basolateral amygdala revealed transcriptional divergence between the male subgroups, including genes with reported polymorphic association to patients with PTSD., Conclusions: This SEFL model provides a tool for development of PTSD therapeutics that is compatible with the growing number of mouse-specific resources. Furthermore, use of an inbred strain allows for investigation into epigenetic mechanisms that are expected to critically regulate susceptibility and resilience., (Copyright © 2017 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)

Desquamative inflammatory vaginitis is a poorly understood chronic vaginitis with an unknown etiology. Symptoms of desquamative inflammatory vaginitis include copious yellowish discharge, vulvovaginal discomfort, and dyspareunia. Cervical ectropion, the presence of glandular columnar cells on the ectocervix, has not been reported as a cause of desquamative inflammatory vaginitis. Although cervical ectropion can be a normal clinical finding, it has been reported to cause leukorrhea, metrorrhagia, dyspareunia, and vulvovaginal irritation. Patients with cervical ectropion and desquamative inflammatory vaginitis are frequently misdiagnosed with candidiasis or bacterial vaginosis and repeatedly treated without resolution of symptoms. We report the case of a 34-year-old woman with a 4-year history of profuse yellowish discharge and dyspareunia. Upon presentation, her symptoms and laboratory results met the criteria for desquamative inflammatory vaginitis, but the standard treatments did not provide long-lasting relief. As a last resort, cryotherapy (cryosurgery) of her cervix was performed for treatment of her cervical ectropion, which provided complete resolution of her symptoms. Mitchell L, King M, Brillhart H, Goldstein A. Cervical Ectropion May Be a Cause of Desquamative Inflammatory Vaginitis. Sex Med 2017;5:e212-e214., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

Global DNA demethylation is an integral part of reprogramming processes in vivo and in vitro, but whether it occurs in the derivation of induced pluripotent stem cells (iPSCs) is not known. Here, we show that iPSC reprogramming involves both global and targeted demethylation, which are separable mechanistically and by their biological outcomes. Cells at intermediate-late stages of reprogramming undergo transient genome-wide demethylation, which is more pronounced in female cells. Global demethylation requires activation-induced cytidine deaminase (AID)-mediated downregulation of UHRF1 protein, and abolishing demethylation leaves thousands of hypermethylated regions in the iPSC genome. Independently of AID and global demethylation, regulatory regions, particularly ESC enhancers and super-enhancers, are specifically targeted for hypomethylation in association with transcription of the pluripotency network. Our results show that global and targeted DNA demethylation are conserved and distinct reprogramming processes, presumably because of their respective roles in epigenetic memory erasure and in the establishment of cell identity., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)

National registration of Australian health practitioners aimed to facilitate workforce mobility. Non-medical prescribers, including nurses, podiatrists and optometrists, are overseen by National Boards which, in some cases, specify a formulary from which their health practitioners may prescribe. All prescribers must comply with their respective State or Territory’s legislation. If prescribing a medicine that is a benefit under the Pharmaceutical Benefits Scheme (PBS), additional restrictions may apply. National Board and PBS prescribing formularies were compared and State and Territory medicines legislation was interrogated regarding non-medical prescribing. Discrepancies were identified between the approved formularies for non-medical prescribers, PBS prescribing formularies and medicines allowed to be prescribed under jurisdictional legislation. Asynchronous medicines legislation provides potential for health professionals to either inadvertently or knowingly breach legislation following national changes to health policy. Consideration should be given to the development of consistent legislation and its uniform commencement across all Australian jurisdictions.

Background: The identification, management, and reporting of adverse drug reactions are integral to clinical practice and education; however, undergraduate teaching related to adverse drug reactions may be inadequate for practice. Existing methods of causality assessment have a number of limitations in relation to clinical teaching, for example, they do not deal well with the concurrent use of other medications., Objective: To develop and pilot a teaching tool to guide students through the process of identifying and prioritizing potential causes of an adverse drug reaction., Setting: University-based School of Pharmacy, Australia: an undergraduate Quality Use of Medicines course., Method: A contrived acronym (mnemonic) was developed from causality assessments and discussions with practitioners. The acronym ATTEND DR (abnormality, taken, timeline, evidence, nothing else?, dose, dechallenge, and rechallenge) was piloted in workshops that focussed on adverse drug reactions and their management. Students' responses to "What did you find most valuable about today's workshop?" and "How could we improve?" were analyzed., Results: All attendees responded (65/65). Students indicated that the ATTEND DR acronym was easy to remember, and facilitated causality assessment in a clinical context, due to an easily followed, step-by-step, comprehensive process that was easy to remember. More practice case studies were requested., Conclusion: The ATTEND DR acronym was designed to address limitations of the existing methods of causality assessment in relation to clinical teaching and preparation of students for future clinical roles. Students responded favorably to its introduction, commenting that it was easily remembered and provided a comprehensive, clinically orientated, step-by-step process., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: There is a need to ascertain the type and level of treatment burden experienced by people with co-morbidities. This is important to identify the characteristics of participants who are at most risk of treatment burden., Objective: The aim of this study is to identify the characteristics of participants who are at most risk of treatment burden., Methods: This cross-sectional study was part of a larger project and recruitment was conducted across four Australian regions: rural, semi-rural and metropolitan. Participants were asked about their treatment burden using an adapted version of a measure, which included the following five dimensions: medication, time and administrative, lifestyle change, social life and financial burden., Results: In total, 581 participants with various chronic health conditions reported a mean global treatment burden of 56.5 out of 150 (standard deviation = 34.5). Number of chronic conditions (β = .34, p < 0.01), age, (β = -.27, p < 0.01), the presence of an unpaid carer (β = .22, p < 0.001) and the presence of diabetes mellitus and other endocrine conditions (β = .13, p < 0.01) were significant predictors of overall treatment burden. For the five dimensions of treatment burden, social, medicine and administrative burden were predicted by the same cluster of variables: number of conditions, age, presence of an unpaid carer and diabetes. However, in addition to these variables, financial dimensions were also predicted by education level, ethnicity and health insurance. Educational level also influenced lifestyle burden., Conclusion: A substantial proportion of community-dwelling adults with chronic conditions have considerable levels of treatment burden. Specifically, health professionals should provide greater focus on managing overall treatment burden for persons who are of young age, have an endocrine condition or an unpaid carer, or a combination of these factors.

Introduction: Ospemifene (Osphena, Shionogi Inc, Florham, NJ, USA) is an estrogen agonist and antagonist approved by the U.S. Federal Drug Administration for the treatment of "moderate to severe dyspareunia, a symptom of vulvar and vaginal atrophy, due to menopause." Although published, peer-reviewed, placebo-controlled studies have shown objective improvement in dyspareunia and in vaginal atrophy, there are no published data that have assessed changes in vulvar atrophy after the use of ospemifene., Aim: To present two cases of women with severe vulvar atrophy that showed no improvement with the use of ospemifene., Methods: A review of two recent cases of a clinic specializing in the treatment of vulvovaginal disorders was performed. Case 1 was a 53-year-old menopausal woman who presented with non-provoked vulvar and vaginal discomfort and introital dyspareunia. She had used ospemifene 60 mg/d for 1.5 years without improvement in her symptoms before presentation. Case 2 was a 57-year-old menopausal woman who also presented with non-provoked vulvar rawness, burning, irritation, vaginal dryness, and introital dyspareunia. She had started ospemifene 60 mg/d 1 year before presentation and reported mild improvement in her vaginal dryness but no improvement in her vulvar irritation or introital dyspareunia., Main Outcome Measures: Change in vulvar atrophy and introital dyspareunia., Conclusion: These cases highlight the need to perform additional clinical trials that specifically assess the efficacy of ospemifene for changes in vulvar atrophy., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction The Nominal Group Technique (NGT) and Delphi Technique are consensus methods used in research that is directed at problem-solving, idea-generation, or determining priorities. While consensus methods are commonly used in health services literature, few studies in pharmacy practice use these methods. This paper provides an overview of the NGT and Delphi technique, including the steps involved and the types of research questions best suited to each method, with examples from the pharmacy literature. Methodology The NGT entails face-to-face discussion in small groups, and provides a prompt result for researchers. The classic NGT involves four key stages: silent generation, round robin, clarification and voting (ranking). Variations have occurred in relation to generating ideas, and how 'consensus' is obtained from participants. The Delphi technique uses a multistage self-completed questionnaire with individual feedback, to determine consensus from a larger group of 'experts.' Questionnaires have been mailed, or more recently, e-mailed to participants. When to use The NGT has been used to explore consumer and stakeholder views, while the Delphi technique is commonly used to develop guidelines with health professionals. Method choice is influenced by various factors, including the research question, the perception of consensus required, and associated practicalities such as time and geography. Limitations The NGT requires participants to personally attend a meeting. This may prove difficult to organise and geography may limit attendance. The Delphi technique can take weeks or months to conclude, especially if multiple rounds are required, and may be complex for lay people to complete.

Grounded theory is a qualitative research methodology that aims to explain social phenomena, e.g. why particular motivations or patterns of behaviour occur, at a conceptual level. Developed in the 1960s by Glaser and Strauss, the methodology has been reinterpreted by Strauss and Corbin in more recent times, resulting in different schools of thought. Differences arise from different philosophical perspectives concerning knowledge (epistemology) and the nature of reality (ontology), demanding that researchers make clear theoretical choices at the commencement of their research when choosing this methodology. Compared to other qualitative methods it has ability to achieve understanding of, rather than simply describing, a social phenomenon. Achieving understanding however, requires theoretical sampling to choose interviewees that can contribute most to the research and understanding of the phenomenon, and constant comparison of interviews to evaluate the same event or process in different settings or situations. Sampling continues until conceptual saturation is reached, i.e. when no new concepts emerge from the data. Data analysis focusses on categorising data (finding the main elements of what is occurring and why), and describing those categories in terms of properties (conceptual characteristics that define the category and give meaning) and dimensions (the variations within properties which produce specificity and range). Ultimately a core category which theoretically explains how all other categories are linked together is developed from the data. While achieving theoretical abstraction in the core category, it should be logical and capture all of the variation within the data. Theory development requires understanding of the methodology not just working through a set of procedures. This article provides a basic overview, set in the literature surrounding grounded theory, for those wanting to increase their understanding and quality of research output.

Critical roles for DNA methylation in embryonic development are well established, but less is known about its roles during trophoblast development, the extraembryonic lineage that gives rise to the placenta. We dissected the role of DNA methylation in trophoblast development by performing mRNA and DNA methylation profiling of Dnmt3a/3b mutants. We find that oocyte-derived methylation plays a major role in regulating trophoblast development but that imprinting of the key placental regulator Ascl2 is only partially responsible for these effects. We have identified several methylation-regulated genes associated with trophoblast differentiation that are involved in cell adhesion and migration, potentially affecting trophoblast invasion. Specifically, trophoblast-specific DNA methylation is linked to the silencing of Scml2, a Polycomb Repressive Complex 1 protein that drives loss of cell adhesion in methylation-deficient trophoblast. Our results reveal that maternal DNA methylation controls multiple differentiation-related and physiological processes in trophoblast via both imprinting-dependent and -independent mechanisms., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Evidence of increased reactive oxygen species (ROS) production is observed in the circulation during exercise in humans. This is exacerbated at elevated body temperatures and attenuated when normal exercise-induced body temperature elevations are suppressed. Why ROS production during exercise is temperature dependent is entirely unknown. This review covers the human exercise studies to date that provide evidence that oxidant and antioxidant changes observed in the blood during exercise are dependent on temperature and fluid balance. We then address possible mechanisms linking exercise with these variables that include shear stress, effects of hemoconcentration, and signaling pathways involving muscle osmoregulation. Since pathways of muscle osmoregulation are rarely discussed in this context, we provide a brief review of what is currently known and unknown about muscle osmoregulation and how it may be linked to oxidant production in exercise and hyperthermia. Both the circulation and the exercising muscle fibers become concentrated with osmolytes during exercise in the heat, resulting in a competition for available water across the muscle sarcolemma and other tissues. We conclude that though multiple mechanisms may be responsible for the changes in oxidant/antioxidant balance in the blood during exercise, a strong case can be made that a significant component of ROS produced during some forms of exercise reflect requirements of adapting to osmotic challenges, hyperthermia challenges, and loss of circulating fluid volume., (Copyright © 2016 the American Physiological Society.)

Unlabelled: Pseudomonas aeruginosa is an opportunistic human pathogen that causes severe, life-threatening infections in patients with cystic fibrosis (CF), endocarditis, wounds, or artificial implants. During CF pulmonary infections, P. aeruginosa often encounters environments where the levels of calcium (Ca(2+)) are elevated. Previously, we showed that P. aeruginosa responds to externally added Ca(2+) through enhanced biofilm formation, increased production of several secreted virulence factors, and by developing a transient increase in the intracellular Ca(2+) level, followed by its removal to the basal submicromolar level. However, the molecular mechanisms responsible for regulating Ca(2+)-induced virulence factor production and Ca(2+) homeostasis are not known. Here, we characterized the genome-wide transcriptional response of P. aeruginosa to elevated [Ca(2+)] in both planktonic cultures and biofilms. Among the genes induced by CaCl2 in strain PAO1 was an operon containing the two-component regulator PA2656-PA2657 (here called carS and carR), while the closely related two-component regulators phoPQ and pmrAB were repressed by CaCl2 addition. To identify the regulatory targets of CarSR, we constructed a deletion mutant of carR and performed transcriptome analysis of the mutant strain at low and high [Ca(2+)]. Among the genes regulated by CarSR in response to CaCl2 are the predicted periplasmic OB-fold protein, PA0320 (here called carO), and the inner membrane-anchored five-bladed β-propeller protein, PA0327 (here called carP). Mutations in both carO and carP affected Ca(2+) homeostasis, reducing the ability of P. aeruginosa to export excess Ca(2+). In addition, a mutation in carP had a pleotropic effect in a Ca(2+)-dependent manner, altering swarming motility, pyocyanin production, and tobramycin sensitivity. Overall, the results indicate that the two-component system CarSR is responsible for sensing high levels of external Ca(2+) and responding through its regulatory targets that modulate Ca(2+) homeostasis, surface-associated motility, and the production of the virulence factor pyocyanin., Importance: During infectious disease, Pseudomonas aeruginosa encounters environments with high calcium (Ca(2+)) concentrations, yet the cells maintain intracellular Ca(2+) at levels that are orders of magnitude less than that of the external environment. In addition, Ca(2+) signals P. aeruginosa to induce the production of several virulence factors. Compared to eukaryotes, little is known about how bacteria maintain Ca(2+) homeostasis or how Ca(2+) acts as a signal. In this study, we identified a two-component regulatory system in P. aeruginosa PAO1, termed CarRS, that is induced at elevated Ca(2+) levels. CarRS modulates Ca(2+) signaling and Ca(2+) homeostasis through its regulatory targets, CarO and CarP. The results demonstrate that P. aeruginosa uses a two-component regulatory system to sense external Ca(2+) and relays that information for Ca(2+)-dependent cellular processes., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Introduction: Lichen sclerosus (LS) is a chronic inflammatory dermatosis, usually affecting the anogenital skin in women. This chronic inflammation can cause scarring of genitalia including narrowing of the introitus and phimosis of the clitoris. These architectural changes can lead to recurrent tearing during intercourse (vulvar granuloma fissuratum) and decreased clitoral sensation. Surgical correction of vulvar granuloma fissuratum (VGF) and clitoral phimosis can be performed, but there is little data on the patient satisfaction and complications following these surgical procedures., Aim: To evaluate patient experience and outcomes in women undergoing surgical correction of scarring caused by anogenital LS., Methods: A retrospective chart review of patients at a vulvar disorders clinic was performed to identify women who had undergone surgical correction of clitoral phimosis or lysis of vulvar adhesions for VGF due to LS. Twenty-eight women were contacted via telephone between 4 and 130 months postoperatively. An eight-question survey was used to determine patient experience and outcomes., Main Outcome Measures: All participants completed an eight-question survey to evaluate patient satisfaction with the surgery, effects on clitoral sensation, orgasm and pain with intercourse, postoperative symptoms or complications, and the presence of recurrent vulvar scarring., Results: Participants reported that they were either very satisfied (44%) or satisfied (40%) with the procedure. Of the women who experienced decreased clitoral sensation prior to surgery, 75% endorsed increased clitoral sensitivity postoperatively. Of the women who had dyspareunia prior to surgery, the majority of women reported having pain-free sex (33%) or improved but not completely pain-free sex (58%) after surgery. There were no complications or symptoms made worse by the surgical procedures., Conclusions: This study shows high patient satisfaction and low complication risk associated with surgical correction of clitoral phimosis and lysis of vulvar adhesions for VGF caused by LS. Patients reported improvement in clitoral sensation and ability to achieve orgasm, as well as decreased dyspareunia. Surgical correction of vulvar scarring is a viable option to restore vulvar anatomy and sexual function in appropriate candidates with anogenital LS. Flynn AN, King M, Rieff M, Krapf J, and Goldstein AT. Patient satisfaction of surgical treatment of clitoral phimosis and labial adhesions caused by lichen sclerosus. Sex Med 2015;3:251-255.

Background: Australian community pharmacies are experiencing challenges, including government prescription pricing reform and a dramatically increasing competitive environment. Enacting appropriate responsive actions requires capable pharmacy managers. 'Capability' implies managing effectively in the present, but with unknown or emerging contexts and with new problems. A conceptual understanding of managerial capability as practiced by pharmacist owner-managers is unavailable in the literature., Objectives: This research aimed to address the question: How can we understand managerial capability in relation to effective community pharmacy management? The study's objective was to develop preliminary theoretical departure points for continuing research responding to the research question., Methods: The objective was approached by exploring how 5 top Australian pharmacy owner-managers accomplish the management of their businesses in a changing business environment. Qualitative research methods were employed to develop a social process perspective of how the managers enact their management practices. In-depth semi-structured life-world interviews were undertaken as the major method of data collection. Interview text thematic analysis was carried out identifying rich conceptual properties and dimensions, which 'dimensionalized' 3 key integrated categories., Results: The findings show how the managers are immersed in their business, managerial and personal practices in a holistic and relational manner. Managerial processes, reported through three conceptual categories, their properties and dimensions, reveal the highly situational nature of the reality the managers were experiencing, including their need to express their personal/professional identity. The properties and dimensions of the category 'learning generatively' in particular, reveal how the pharmacy owner-managers shape their business activities and their emerging context as time passes., Conclusions: The preliminary interpretive view of managerial capability describes the phenomenon as an emergent human accomplishment rather than a possessed ability. This social process perspective enables the inclusion of context with time. The study acts as a formative departure point for continuing research of pharmacist managerial capability which seeks to better understand the linkage between 'knowing' and 'doing'., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Context: Treatment burden, the burden associated with the treatment and management of chronic illness, has not yet been well articulated., Objective: Using Rodgers' (1989, Journal of Advanced Nursing, 14, 330-335) method of concept analysis, this review describes the ways in which treatment burden has been conceptualized to define the concept and to develop a framework for understanding its attributes, antecedents and consequences., Methods: Leading databases were searched electronically between the years 2002 and 2011. To ensure the review focused on actual observations of the concept of interest, articles that did not measure treatment burden (either qualitatively or quantitatively) were excluded. An inductive approach was used to identify themes related to the concept of treatment burden., Main Results: Thirty articles, identified from 1557 abstracts, were included in the review. The attributes of treatment burden include burden as a dynamic process, as a multidimensional concept, and comprising of both subjective and objective elements. Prominent predisposing factors (antecedents) include the person's age and gender, their family circumstances, possible comorbidity, high use of medications, characteristics of treatment and their relationship with their health-care provider. The most dominant consequences are poor health and well-being, non-adherence to treatment, ineffective resource use and burden on significant others. Furthermore, many of these consequences can also become antecedents, reflecting the cyclic and dynamic nature of treatment burden., Conclusion: The findings underscore the need for researchers and health-care professionals to engage in collaborative discussions and make cooperative efforts to help alleviate treatment burden and tailor treatment regimens to the realities of people's daily lives., (© 2013 Blackwell Publishing Ltd.)

In successful community pharmacy business enterprises suitably responsive actions to meet ever-increasing change require capable pharmacy managers who readily learn and adapt. Capability as a concept is generally understood to be the ability of a manager to identify and act to solve unfamiliar problems in unfamiliar situations. Capability is characterized by adaptability and flexibility. However, different understandings of the concept 'capability' and what it means to be 'capable' are indirect and incomplete. This paper aims to clarify current theories regarding the concept of 'capability' at the level of the individual, and through this to make more explicit what is known about the phenomenon, but more particularly, how we know what we know. The analysis includes the concept of 'competence' because explanations of capability include competence, and the two concepts are not clearly separated in the literature. By probing the epistemological origins of current theory concerning both concepts, the limiting taken for granted assumptions are revealed. Assumptions about context and time, and the psychological theory through which individuals are assumed to perceive, know and learn, are illuminated. The analysis, in connection with the literature, shows how the interpretive philosophic research approach may reveal a different and useful theoretical perspective for explaining capability as a dynamic performance. It is suggested that such a perspective may narrow the gap between the theory of capability and its practice. The interpretive perspective holds potential to reveal how capability, as performed by successful community pharmacy managers, might be further researched and strengthened. This paper supports the challenging suggestion that pharmacy social research needs to rebalance the dominance of purely empirical research by exploring interpretive methodologies to better understand human actions and relations in the context of pharmacy., (Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.)

Background: Pregabalin is a treatment option for patients with persistent neuropathic pain. Its use has been associated with changes in mood and the development of depression and/or suicidal ideation., Objective: Case presentations were reviewed of five patients reporting changes in mood, depression and suicidal ideation from the first 50 (approximately) patients commenced on pregabalin at the clinic., Discussion: Although these patients had a history of depression, their mood had been stable before commencing pregabalin. Soon after commencement they reported changes in mood, and development of depression and/or suicidal ideation, which improved with dose reduction or cessation of pregabalin. Ultimately, all five patients ceased pregabalin treatment. Suicidal ideation is a recognised adverse effect of pregabalin. Patients should be warned of and monitored for deterioration in mood.

Objective: To explore the attributes of pharmacy choice for people with chronic conditions., Methods: Semi-structured interviews were conducted between May and October 2012, across four regions in three Australian states. Purposive sampling was used to recruit participants with chronic conditions and unpaid carers. Interviews were analysed via the constant comparison method., Key Findings: Ninety-seven interviews were conducted. The majority of participants were regular patrons of one pharmacy and five attributes influenced this choice: patient-centred care, convenience, price, personal trait or preference and service/medication need. Patient-centred care, such as providing individualised medication counselling, continuity of care, development of relationships and respectful advice, emerged as an important attribute. There was minimal discussion as to choosing a pharmacy based on the provision of professional services, underscoring the limited consumer knowledge of such services and related standards of care., Conclusion: Patient-centred care is an important attribute of quality care as perceived by people who are regular community pharmacy users. These findings highlight the need for pharmacy staff to implement a patient-centred approach to care, thus meeting the perceived needs of their customers. A greater effort is also necessary to raise the profile of pharmacy as a healthcare destination., (© 2013 Royal Pharmaceutical Society.)

Objectives: To explore pharmacy students' ethical behaviour and care towards patients in relation to the provision of emergency hormonal contraception (EHC)., Methods: Three hundred and forty-seven pharmacy students were presented a hypothetical scenario involving refusal of EHC, based on religious or moral grounds, and asked to write responses as to how the patient should be managed; 270 (77.8%) responded., Key Findings: Of all respondents, 90.4% referred the patient to another health professional to facilitate continuity of care, with referrals increasing as students progressed through the programme. Religion had no influence on referral, while female gender was related to increased referral., Conclusions: Gender difference, if continued into practice, has the potential to negatively impact on patient care., (© 2013 Royal Pharmaceutical Society.)