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The aim of this paper is to contribute to a better understanding of musician identity learning. Identity is a significant driving-force behind many artistic vocations. However, identity may also pose challenges such as vulnerability, burnout and conflict of commitment. The paper focuses on musician identity, based on interviews with classical and rock musicians during the COVID-19 pandemic. Analyses reveal that the musicians are either in integrating or situating musician identity learning, two different processes discussed as dynamically interrelated and depending on life situation. The paper argues that breaks of normality accentuate challenges stemming from musician identity and that the proposed learning processes are responses to those challenges, which is previously under-explored. The paper concludes that many musicians or other artistic practitioners likely struggle to de-centralize their identity in order to foster a sustainable vocational career and personal growth, and that temporary career breaks may provide necessary relief and learning space.

Tanja Sofie Hansen,1,2 Ingrid Poulsen,2– 4 Vibeke Nørholm,3 Mia Ingerslev Loft,4,5 Pia Søe Jensen3,4,6 1Department of Medical Diseases, Amager Hvidovre Copenhagen University Hospital, Copenhagen, Denmark; 2Department of Public Health, Section of Nursing, Aarhus University, Aarhus, Denmark; 3Department of Clinical Research, Hvidovre Copenhagen University Hospital, Copenhagen, Denmark; 4Institute for People and Technology, Roskilde University, Roskilde, Denmark; 5Department of Neurology, Rigshospitalet, Copenhagen, Denmark; 6Department of Orthopaedic Surgery, Hvidovre Copenhagen University Hospital, Copenhagen, DenmarkCorrespondence: Tanja Sofie Hansen, Amager Hvidovre Copenhagen University Hospital, Kettegaard Allé 30, Hvidovre, 2650, Denmark, Tel +45 30 48 89 30, Email tanja.sofie.hansen@regionh.dkBackground: The knowledge is sparse in the literature on intervention programs using nutritional support and physical activity for patients with chronic obstructive pulmonary disease within a person-centred approach. We aimed to explore and map the existing evidence on intervention programs with a person-centred approach, focusing on nutritional support and physical activity for people with COPD.Methods: A scoping review was conducted using Arksey & O’Malley’s methodological framework. A search in the databases CINAHL and PubMed, and a grey literature search, was conducted in June 2022 and updated in June 2023. We identified studies published between 2012 and 2023. The PRISMA checklist for scoping reviews, supported by The PAGER framework was used for reporting the method.Results: A total of 15 studies were included. The primary interventions comprised behavior of change or self-management, addressing needs assessment, motivation, personal goals, education, and physical activity. Health-related quality of life and hospital stay displayed no clinically significant variances. However, eight studies demonstrated differences in physical function and activity levels. Nutritional outcomes were addressed in one study, and three studies involved relatives.Conclusion: This scoping review addresses a knowledge gap in nutritional support interventions with a person-centred approach. It indicates that there is a need to increase nutritional support and consider the patient’s physical and social environmental resources within Behavior of change or Self-management intervention programs for patients with COPD. The review found no clinical effect on health-related quality of life, although there were some effects on physical activity. The results highlight how the interdisciplinary team can include the patients’ resources when structuring the management of COPD by applying a person-centred approach.Keywords: COPD, pulmonary rehabilitation, self-management program, motivational techniques

Abstract Thermostable DNA polymerases, such as Taq isolated from the thermophilic bacterium Thermus aquaticus, enable one-pot exponential DNA amplification known as polymerase chain reaction (PCR). However, properties other than thermostability - such as fidelity, processivity, and compatibility with modified nucleotides - are important in contemporary molecular biology applications. Here, we describe the engineering and characterization of a fusion between a DNA polymerase identified in the marine archaea Nanoarchaeum equitans and a DNA binding domain from the thermophile Sulfolobus solfataricus. The fusion creates a highly active enzyme, Neq2X7, capable of amplifying long and GC-rich DNA, unaffected by replacing dTTP with dUTP in PCR, and tolerant to various known PCR inhibitors. This makes it an attractive DNA polymerase for use, e.g., with uracil excision (USER) DNA assembly and for contamination-free diagnostics. Using a magnification via nucleotide imbalance fidelity assay, Neq2X7 was estimated to have an error rate lower than 2 ∙ 10−5 bp−1 and an approximately 100x lower fidelity than the parental variant Neq2X, indicating a trade-off between fidelity and processivity – an observation that may be of importance for similarly engineered DNA polymerases. Neq2X7 is easy to produce for routine application in any molecular biology laboratory, and the expression plasmid is made freely available.

The global pandemic of Covid-19 has had massive effects on many levels. For many sectors, the restrictions are gone but normalcy is not fully restored. The performing arts, for example, have been especially vulnerable to sharp restrictions, with audiences staying at home. The aim of this article is to explore what it means for freelance musicians in the art/classical music genre to re-establish their careers after the pandemic. The empirical data consists of interviews with Swedish freelance musicians from 2021 and 2022. The article concludes that the pandemic and its aftermath present the freelancers with substantial practical challenges and emotional dilemmas. Some of the freelancers have been able to re-establish their careers and performer identities, while others are still struggling to get back into the loop. The pandemic has started processes of change, which are now negotiated in relation to going back to ‘normal’.

In adipose tissue, reduced expression of the glycerol channel aquaporin 7 (AQP7) has been associated with increased accumulation of triglyceride. The present study determines the relative protein abundances of lipolytic enzymes, AQP7, and cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) in paired mesenteric and omental visceral adipose tissue (VAT) and abdominal and femoral subcutaneous adipose tissue (SAT) in women with either normal weight or upper-body obesity. No differences in the expression of hormone-sensitive lipase (HSL) or AQP7 were found between the two groups in the four depots. The expression of adipocyte triglyceride lipase (ATGL) and HSL were higher in omental VAT and femoral SAT than in mesenteric VAT in both groups of women. Similarly, AQP7 expression was higher in omental VAT than in mesenteric VAT. The expression of PEPCK-C was lower in omental VAT than in femoral SAT. No correlation between the expression of AQP7 and the mean adipocyte size was observed; however, the expression of PEPCK-C positively correlated with the mean adipocyte size. In conclusion, a depot-specific protein expression pattern was found for ATGL, HSL, AQP7, and PEPCK-C. The expression pattern supports that the regulation of AQP7 protein expression is at least in part linked to the lipolytic rate. Furthermore, the results support that the synthesis of glycerol-3-phosphate via glyceroneogenesis contributes to regulating triglyceride accumulation in white adipose tissue in women.

Abstract Lytic polysaccharide monooxygenases (LPMOs) are oxidative enzymes that help break down lignocellulose, making them highly attractive for improving biomass utilization in industrial biotechnology. The catalytically essential N-terminal histidine (His1) of LPMOs is post-translationally modified by methylation in filamentous fungi to protect them from auto-oxidative inactivation, however, the responsible methyltransferase enzyme is unknown. Using mass-spectrometry-based quantitative proteomics in combination with systematic CRISPR/Cas9 knockout screening in Aspergillus nidulans, we identify the N-terminal histidine methyltransferase (NHMT) encoded by the gene AN4663. Targeted proteomics confirm that NHMT was solely responsible for His1 methylation of LPMOs. NHMT is predicted to encode a unique seven-transmembrane segment anchoring a soluble methyltransferase domain. Co-localization studies show endoplasmic reticulum residence of NHMT and co-expression in the industrial production yeast Komagataella phaffii with LPMOs results in His1 methylation of the LPMOs. This demonstrates the biotechnological potential of recombinant production of proteins and peptides harbouring this specific post-translational modification.

Charlotte Sandau,1 Ejvind Frausing Hansen,1 Thomas Jørgen Ringbæk,2 Thomas Kallemose,3 Dorthe Gaby Bove,4 Ingrid Poulsen,3,5 Vibeke Nørholm,3 Lars Pedersen,6 Jens Ulrik Stæhr Jensen,7,8 Charlotte Suppli Ulrik1,9 1Department of Respiratory Medicine and Endocrinology, Pulmonary Section, Copenhagen University Hospital Hvidovre, Hvidovre, Denmark; 2Allergy and Lung Clinic, Elsinore, Denmark; 3Department of Clinical Research, Copenhagen University Hospital Hvidovre, Hvidovre, Denmark; 4University College Absalon, Centre for Nursing, Roskilde, Denmark; 5Research Unit Nursing and Health Care, Aarhus University, Aarhus, Denmark; 6Department of Respiratory Medicine and Infectious Diseases, Copenhagen University Bispebjerg Hospital, Copenhagen, Denmark; 7Respiratory Medicine Section, Department of Medicine, Herlev-Gentofte Hospital, Hellerup, Denmark; 8Department of Clinical Medicine, Faculty of Health Sciences, University of Copenhagen, København, Denmark; 9Institute of Clinical Medicine, University of Copenhagen, Copenhagen, DenmarkCorrespondence: Charlotte Sandau, Email charlotte.sandau.bech.01@regionh.dkObjective: Devices for Automated Oxygen Administration (AOA) have been developed to optimize the therapeutic benefit of oxygen supplementation. We aimed to investigate the effect of AOA on multidimensional aspects of dyspnea and as-needed consumption of opioids and benzodiazepines, as opposed to conventional oxygen therapy, in hospitalized patients with Acute Exacerbation of COPD (AECOPD).Method and Patients: A multicenter randomized controlled trial across five respiratory wards in the Capital Region of Denmark. Patients admitted with AECOPD (n=157) were allocated 1:1 to either AOA (O2matic Ltd), a closed loop device automatically delivering oxygen according to the patient’s peripheral oxygen saturation (SpO2), or conventional nurse-administered oxygen therapy. Oxygen flows and SpO2 levels were measured by the O2matic device in both groups, while dyspnea, anxiety, depression, and COPD symptoms were accessed by Patient Reported Outcomes.Results: Of the 157 randomized patients, 127 had complete data for the intervention. The AOA reduced patients’ perception of overall unpleasantness significantly on the Multidimensional Dyspnea Profile (MDP) with a difference in medians of − 3 (p=0.003) between the intervention group (n=64) and the control group (n=63). The AOA also provided a significant between group difference in all single items within the sensory domain of the MDP (all p-values≤ 0.05) as well as in the Visual Analogue Scale – Dyspnea (VAS-D) within the past three days (p=0.013). All between group differences exceeded the Minimal Clinical Important Difference of the MDP and VAS-D, respectively. AOA did not seem to have an impact on the emotional response domain of the MDP, the COPD Assessment Test, the Hospital Anxiety and Depression Scale, or use of as-needed opioids and/or benzodiazepines (all p-values> 0.05).Conclusion: AOA reduces both breathing discomfort and physical perception of dyspnea in patients admitted with AECOPD but did not seem to impact the emotional status or other COPD symptoms.Keywords: dyspnea, oxygen, automated oxygen therapy, multidimensional dyspnea profile, COPD, hospital, admission, therapy

Recently, health technology systems offering monitoring of the peripheral oxygen saturation level and automated oxygen administration (AOA) have emerged. AOA has been shown to reduce duration of hypoxemia and the length of hospital stay, but the patients’ perspective on AOA has not been investigated. This qualitative study, based on the interpretive description methodology, aimed to explore how patients hospitalized with exacerbation of chronic obstructive pulmonary disease (COPD) experience being treated with AOA. Eighteen patients treated with AOA were included in the study. Data was collected during admission or in the patients’ homes using semi-structured interviews focusing on patients’ experiences of AOA using the word “robot” as used by patients. The findings revealed two themes “adaptation of behavior to the robot” and “robots can make patients feel safe but not cared for” and six subthemes. Our findings illustrate how patients were willing to compromise their own therapy and thereby safety by avoiding behavior triggering AOA alarms and disturbing their fellow patients and the health care professionals. Adherence, defined as patients’ consistency in taking their medications as prescribed, becomes an important point of attention for health professionals when applying individualized robotic therapies such as AOA to patients with COPD. To support patients in the process of managing adherence to therapeutic technology, we propose a person-centered care approach that, through education and communication with the patients, generates an understanding of how they can self-manage AOA and its alarms without activating avoiding behavior that threatens their treatment and recovery.

The recently discovered lytic polysaccharide monooxygenases (LPMOs) are Cu-containing enzymes capable of degrading polysaccharide substrates oxidatively. The generally accepted first step in the LPMO reaction is the reduction of the active-site metal ion from Cu2+ to Cu+. Here we have used a systematic diffraction data collection method to monitor structural changes in two AA9 LPMOs, one from Lentinus similis (LsAA9_A) and one from Thermoascus aurantiacus (TaAA9_A), as the active-site Cu is photoreduced in the X-ray beam. For LsAA9_A, the protein produced in two different recombinant systems was crystallized to probe the effect of post-translational modifications and different crystallization conditions on the active site and metal photoreduction. We can recommend that crystallographic studies of AA9 LPMOs wishing to address the Cu2+ form use a total X-ray dose below 3 × 104 Gy, while the Cu+ form can be attained using 1 × 106 Gy. In all cases, we observe the transition from a hexacoordinated Cu site with two solvent-facing ligands to a T-shaped geometry with no exogenous ligands, and a clear increase of the θ2 parameter and a decrease of the θ3 parameter by averages of 9.2° and 8.4°, respectively, but also a slight increase in θT. Thus, the θ2 and θ3 parameters are helpful diagnostics for the oxidation state of the metal in a His-brace protein. On binding of cello-oligosaccharides to LsAA9_A, regardless of the production source, the θT parameter increases, making the Cu site less planar, while the active-site Tyr—Cu distance decreases reproducibly for the Cu2+ form. Thus, the θT increase found on copper reduction may bring LsAA9_A closer to an oligosaccharide-bound state and contribute to the observed higher affinity of reduced LsAA9_A for cellulosic substrates.

Studies of sexual harassment in professional contexts, including academia, provide detailed explanations of the predominance and pervasiveness of sexist organizational norms that enable harassing behavior—and offer a thorough critique of the structures and practices that support and reproduce these norms. When sexist organizational norms are linked to acts of sexual harassment, it becomes clear that harassment is systemic, and that organizations tend to justify and excuse the very norms and behaviors that propagate harassment. Focusing on the context of Danish universities, we do not ask whether sexism exists in Danish society generally and in academia specifically, but rather, why issues of systemic sexism and normalized sexual harassment have been ignored for so long and how sexist organizational norms have been maintained. Based on an investigation of prevalent rhetorical strategies for legitimating sexual harassment and gendered discrimination, we discuss how recognizing these strategies may translate into concerted action against them. Introducing queer organization studies as a lever for such translation, we suggest that a norm‐critical approach may, first, explain how currently dominant norms offer sexist excuses for continued harassment and, consequently, delegitimize and change these unjust norms and the untenable practices they support. [ABSTRACT FROM AUTHOR]

Background and objectiveLiver cirrhosis represents a considerable health burden and causes 1.2 million deaths annually. Patients with decompensated liver cirrhosis have a poor prognosis and severely reduced health-related quality of life. Nurse-led outpatient care has proven safe and feasible for several chronic diseases and engaging nurses in the outpatient care of patients with liver cirrhosis has been recommended. At the decompensated stage, the treatment and nursing care are directed at specific complications, educational support, and guidance concerning preventive measures and signs of decompensation. This review aimed to assess the effects of nurse-assisted follow-up after admission with decompensation in patients with liver cirrhosis from all causes.MethodA systematic search was conducted through February 2022. Studies were eligible for inclusion if i) they assessed adult patients diagnosed with liver cirrhosis that had been admitted with one or more complications to liver cirrhosis and ii) if nurse-assisted follow-up, including nurse-assisted multidisciplinary interventions, was described in the manuscript. Randomized clinical trials were prioritized, but controlled trials and prospective cohort studies with the intervention were also included. Primary outcomes were mortality and readmission, but secondary subjective outcomes were also assessed.Results and conclusionWe included eleven controlled studies and five prospective studies with a historical control group comprising 1224 participants. Overall, the studies were of moderate to low quality, and heterogeneity across studies was substantial. In a descriptive summary, the 16 studies were divided into three main types of interventions: educational interventions, case management, and standardized hospital follow-up. We saw a significant improvement across all types of studies on several parameters, but currently, no data support a specific type of nurse-assisted, post-discharge intervention. Controlled trials with a predefined intervention evaluating clinically- and practice-relevant endpoints in a real-life, patient-oriented setting are highly warranted.

Summary Secreted proteins and peptides hold large potential both as therapeutics and as enzyme catalysts in biotechnology. The high stability of many secreted proteins helps maintain functional integrity in changing chemical environments and is a contributing factor to their commercial potential. Disulphide bonds constitute an important post‐translational modification that stabilizes many of these proteins and thus preserves the active state under chemically stressful conditions. Despite their importance, the discovery and applications within this group of proteins and peptides are limited by the availability of synthetic biology tools and heterologous production systems that allow for efficient formation of disulphide bonds. Here, we refine the design of two DisCoTune (Disulphide bond formation in E. coli with tunable expression) plasmids that enable the formation of disulphides in the highly popular Escherichia coli T7 protein production system. We show that this new system promotes significantly higher yield and activity of an industrial protease and a conotoxin, which belongs to a group of disulphide‐rich venom peptides from cone snails with strong potential as research tools and pharmacological agents.

Microbial evolution often involves transient phenotypes and sequential development of multiple mutations of unclear relevance. Here, the authors show that the evolution of non-growing E. coli cells can be driven by alterations in pyrimidine nucleoside levels associated with colony ageing and/or due to mutations in metabolic or regulatory genes.

Genome engineering is challenging compared to plasmid DNA manipulation. Here the authors create a simple methodology called SEGA that enables genome engineering by combining DNA and bacterial cells followed by identification of recombinant clones by a change in colour when grown on agar plates.

Heyde and Nørholm investigate the role of RNAse E in the toxicity mediated by overexpression of heterologous proteins in derivatives of the BL21(DE3) strain of E. coli. The authors report an unexpected connection between global RNA metabolism and the efficiency of heterologous protein production in E. coli.

Abstract Poly(ethylene terephthalate) (PET) is the world’s most abundant polyester plastic, and its ongoing accumulation in nature is causing a global environmental problem. Currently, the main recycling processes utilize thermomechanical or chemical means, resulting in the deterioration of the mechanical properties of PET. Consequently, polluting de novo synthesis remains preferred, creating the need for more efficient and bio-sustainable ways to hydrolyze the polymer. Recently, a PETase enzyme from the bacterium Ideonella sakaiensis was shown to facilitate PET biodegradation, albeit at slow rate. Engineering of more efficient PETases is required for industrial relevance, but progress is currently hampered by the dependency on intracellular expression in Escherichia coli. To create a more efficient screening platform in E. coli, we explore different surface display anchors for fast and easy assaying of PETase activity. We show that PETases can be functionally displayed on the bacterial cell surface, enabling screening of enzyme activity on PET microparticles – both while anchored to the cell and following solubilization of the enzymes.

In adipose tissue, reduced expression of the glycerol channel aquaporin 7 (AQP7) has been associated with increased accumulation of triglyceride. The present study determines the relative protein abundances of lipolytic enzymes, AQP7, and cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) in paired mesenteric and omental visceral adipose tissue (VAT) and abdominal and femoral subcutaneous adipose tissue (SAT) in women with either normal weight or upper-body obesity. No differences in the expression of hormone-sensitive lipase (HSL) or AQP7 were found between the two groups in the four depots. The expression of adipocyte triglyceride lipase (ATGL) and HSL were higher in omental VAT and femoral SAT than in mesenteric VAT in both groups of women. Similarly, AQP7 expression was higher in omental VAT than in mesenteric VAT. The expression of PEPCK-C was lower in omental VAT than in femoral SAT. No correlation between the expression of AQP7 and the mean adipocyte size was observed; however, the expression of PEPCK-C positively correlated with the mean adipocyte size. In conclusion, a depot-specific protein expression pattern was found for ATGL, HSL, AQP7, and PEPCK-C. The expression pattern supports that the regulation of AQP7 protein expression is at least in part linked to the lipolytic rate. Furthermore, the results support that the synthesis of glycerol-3-phosphate via glyceroneogenesis contributes to regulating triglyceride accumulation in white adipose tissue in women. [ABSTRACT FROM AUTHOR]

Within a timespan of a few years, regulatory sandboxes, a novel initiative for testing fintech innovations, have spread to financial conduct authorities across the world. We study the discursive diffusion of this regulatory innovation as evidence of a broader reconfiguration of relations between the financial sector and its regulators. To do so, we analyse media articles, policy documents, and political speeches, identifying the logics and arguments used to diffuse rhetorical sandboxes. Based on the initial mapping, we detail the anatomy of three national arguments, showing how the underlying rationales for establishing sandboxes are variously reproduced across the contexts of the UK, Singapore, and Denmark. In each case, we find, innovation is posited as a driver of competition, which, in turn, leads to economic growth. Through regulatory sandboxes, financial regulators are becoming actively involved in the promotion of innovation and, hence, continue to rely on and reproduce the exhausted promissory legitimacy of neoliberalism. Whilst appearing to depoliticize the field of finance, the assumption that regulators should be 'open for business' is in itself political; as such, we argue, it needs to be approached more critically, which becomes possible by shifting the analytical focus from discursive diffusion to rhetorical struggle. [ABSTRACT FROM AUTHOR]

Abstract Background Lytic polysaccharide monooxygenases (LPMOs) are important industrial enzymes known for their catalytic degradation of recalcitrant polymers such as cellulose or chitin. Their activity can be measured by lengthy HPLC methods, while high-throughput methods are less specific. A fast and specific LPMO assay would simplify screening for new or engineered LPMOs and accelerate biochemical characterization. Results A novel LPMO activity assay was developed based on the production of the dye phenolphthalein (PHP) from its reduced counterpart (rPHP). The colour response of rPHP oxidisation catalysed by the cellulose-specific LPMO from Thermoascus aurantiacus (TaAA9A), was found to increase tenfold by adding dehydroascorbate (DHA) as a co-substrate. The assay using a combination of rPHP and DHA was tested on 12 different metallo-enzymes, but only the LPMOs catalysed this reaction. The assay was optimized for characterization of TaAA9A and showed a sensitivity of 15 nM after 30 min incubation. It followed apparent Michaelis–Menten kinetics with k cat = 0.09 s−1 and K M = 244 µM, and the assay was used to confirm stoichiometric copper–enzyme binding and enzyme unfolding at a temperature of approximately 60 °C. DHA, glutathione and fructose were found to enhance LPMO oxidation of rPHP and in the optimized assay conditions these co-substrates also enabled cellulose degradation. Conclusions This novel and specific LPMO assay can be carried out in a convenient microtiter plate format ready for high-throughput screening and enzyme characterization. DHA was the best co-substrate tested for oxidation of rPHP and this preference appears to be LPMO-specific. The identified co-substrates DHA and fructose are not normally considered as LPMO co-substrates but here they are shown to facilitate both oxidation of rPHP and degradation of cellulose. This is a rare example of a finding from a high-throughput assay that directly translate into enzyme activity on an insoluble substrate. The rPHP-based assay thus expands our understanding of LPMO catalysed reactions and has the potential to characterize LPMO activity in industrial settings, where usual co-substrates such as ascorbate and oxygen are depleted.

Abstract Background Recombinant proteins are often engineered with an N-terminal signal peptide, which facilitates their secretion to the oxidising environment of the periplasm (gram-negative bacteria) or the culture supernatant (gram-positive bacteria). A commonly encountered problem is that the signal peptide influences the synthesis and secretion of the recombinant protein in an unpredictable manner. A molecular understanding of this phenomenon is highly sought after, as it could lead to improved methods for producing recombinant proteins in bacterial cell factories. Results Herein we demonstrate that signal peptides contribute to an unpredictable translation initiation region. A directed evolution approach that selects a new translation initiation region, whilst leaving the amino acid sequence of the signal peptide unchanged, can increase production levels of secreted recombinant proteins. The approach can increase production of single chain antibody fragments, hormones and other recombinant proteins in the periplasm of E. coli. Conclusions The study demonstrates that signal peptide performance is coupled to the efficiency of the translation initiation region.