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Opinion polls have documented a considerable public skepticism towards a COVID-19 vaccine. Seeking to address the vaccine skepticism challenge this essay surveys the research on vaccine hesitancy and trust building through the lens of the rhetorical situation and points towards five broad principles for a content strategy for public health communicators in regards to vaccination: 1) vaccine hesitancy is not irrational per se; 2) messages should be tailored to the various hesitancy drivers; 3) what is perceived as trustworthy is situational and constantly negotiated; 4) in areas of uncertainty where no exact knowledge exists, the character of the speaker becomes more important; and 5) the trustworthiness of the speaker can be strengthened through finding some common ground—such as shared feelings or accepted premises—with the audience. Such common insights are on offer in the literature on rhetoric and persuasion and linked here with the research on vaccine communication and trust focusing specifically on the latter and character.

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'. [ABSTRACT FROM AUTHOR]

The Escherichia coli cyclic AMP receptor protein (CRP or catabolite activator protein, CAP) provides a textbook example of bacterial transcriptional regulation and is one of the best studied transcription factors in biology. For almost five decades a large number of mutants, evolved in vivo or engineered in vitro, have shed light on the molecular structure and mechanism of CRP. Here, we review previous work, providing an overview of studies describing the isolation of CRP mutants. Furthermore, we present new data on deep sequencing of different bacterial populations that have evolved under selective pressure that strongly favors mutations in the crp locus. Our new approach identifies more than 100 new CRP mutations and paves the way for a deeper understanding of this fascinating bacterial master regulator. Keywords: cAMP receptor protein, Carbon catabolite repression, Global transcriptional regulation, Experimental evolution

This article concerns learning, socialization and social sorting among successful musicians. The aim is to understand and problematize not only what it means to become a classical musician but also what it entails to be a successful classical musician. In what ways are musicians socialized through life? What approaches are formed among musicians, and what is the meaning of these when it comes to performance of work? Are their specific arenas and rituals of particular importance, when it comes to socializing and sorting among musicians? Interviews and documents about twelve musicians have been analyzed with the aid of Bourdieusian concepts, for example, practical sense and habitus, and research on elitist practices within music, arts and sports. The results indicate that there is a typical and similar career path for successful musicians, where young musicians are socialized into considering their careers in music as natural and given. The musicians are recognized as talented or wonder children at an early age, and they are accepted as students of a master teacher. Their careers are built by merits, such as a spectacular debut leading to an international career. These career steps function as both merits and as an opening of doors. Further, they contribute by strengthening the musicians´ identities and their roles as successful musicians and role models within the field.

Lytic Polysaccharide Monooxygenases (LPMOs) oxidatively cleave recalcitrant polysaccharides. The mechanism involves (i) reduction of the Cu, (ii) polysaccharide binding, (iii) binding of different oxygen species, and (iv) glycosidic bond cleavage. However, the complete mechanism is poorly understood and may vary across different families and even within the same family. Here, we have investigated the protonation state of a secondary co-ordination sphere histidine, conserved across AA9 family LPMOs that has previously been proposed to be a potential proton donor. Partial unrestrained refinement of newly obtained higher resolution data for two AA9 LPMOs and re-refinement of four additional data sets deposited in the PDB were carried out, where the His was refined without restraints, followed by measurements of the His ring geometrical parameters. This allowed reliable assignment of the protonation state, as also validated by following the same procedure for the His brace, for which the protonation state is predictable. The study shows that this histidine is generally singly protonated at the Nε2 atom, which is close to the oxygen species binding site. Our results indicate robustness of the method. In view of this and other emerging evidence, a role as proton donor during catalysis is unlikely for this His.

Abstract Background The market for recombinant proteins is on the rise, and Gram-positive strains are widely exploited for this purpose. Bacillus subtilis is a profitable host for protein production thanks to its ability to secrete large amounts of proteins, and Lactococcus lactis is an attractive production organism with a long history in food fermentation. Results We have developed a synbio approach for increasing gene expression in two Gram-positive bacteria. First of all, the gene of interest was coupled to an antibiotic resistance gene to create a growth-based selection system. We then randomised the translation initiation region (TIR) preceding the gene of interest and selected clones that produced high protein titres, as judged by their ability to survive on high concentrations of antibiotic. Using this approach, we were able to significantly increase production of two industrially relevant proteins; sialidase in B. subtilis and tyrosine ammonia lyase in L. lactis. Conclusion Gram-positive bacteria are widely used to produce industrial enzymes. High titres are necessary to make the production economically feasible. The synbio approach presented here is a simple and inexpensive way to increase protein titres, which can be carried out in any laboratory within a few days. It could also be implemented as a tool for applications beyond TIR libraries, such as screening of synthetic, homologous or domain-shuffled genes.

Danish retail banking is characterized by a role conflict between sales and advice. Through the lens of identity work, this study explores how bankers negotiate this conflict by seeking to make sense of themselves and justify the work that they do. We show that bankers are keenly aware of the role conflicts they face in doing their jobs, but they also discover ways of making these conflicts meaningfully productive for themselves, their employers, and their customers. To explain these findings, we introduce Hartmut Rosa's concept of resonance, provisionally understood as a processual and relational conceptualization of well-being. This leads to a conceptualization of bankers' identity work of managing the sales–advice tension as striving for professional resonance, the attainment of which is possible through three strategies: an advice orientation, which can be understood as a protection of resonance; a sale orientation, which relies on a promise of resonance; and an integration of sales and advice, which can be seen as an incorporation of resonance. Following the analysis, the normativity of resonance theory is leveraged in a critical discussion of the identified strategies of professional resonance, and the potential implications for well-being research and human resource management practices are considered. [ABSTRACT FROM AUTHOR]

Abstract Background Plasmids are widely used and essential tools in molecular biology. However, plasmids often impose a metabolic burden and are only temporarily useful for genetic engineering, bio-sensing and characterization purposes. While numerous techniques for genetic manipulation exist, a universal tool enabling rapid removal of plasmids from bacterial cells is lacking. Results Based on replicon abundance and sequence conservation analysis, we show that the vast majority of bacterial cloning and expression vectors share sequence similarities that allow for broad CRISPR-Cas9 targeting. We have constructed a universal plasmid-curing system (pFREE) and developed a one-step protocol and PCR procedure that allow for identification of plasmid-free clones within 24 h. While the context of the targeted replicons affects efficiency, we obtained curing efficiencies between 40 and 100% for the plasmids most widely used for expression and engineering purposes. By virtue of the CRISPR-Cas9 targeting, our platform is highly expandable and can be applied in a broad host context. We exemplify the wide applicability of our system in Gram-negative bacteria by demonstrating the successful application in both Escherichia coli and the promising cell factory chassis Pseudomonas putida. Conclusion As a fast and freely available plasmid-curing system, targeting virtually all vectors used for cloning and expression purposes, we believe that pFREE has the potential to eliminate the need for individualized vector suicide solutions in molecular biology. We envision the application of pFREE to be especially useful in methodologies involving multiple plasmids, used sequentially or simultaneously, which are becoming increasingly popular for genome editing or combinatorial pathway engineering.

Abstract How do ageing bacterial colonies generate adaptive mutants? Over a period of two months, we isolated on ageing colonies outgrowing mutants able to use a new carbon source, and sequenced their genomes. This allowed us to uncover exquisite details on the molecular mechanism behind their adaptation: most mutations were located in just a few hotspots in the genome, and over time, mutations increasingly were consistent with the involvement of 8-oxo-guanosine, formed exclusively on the transcribed strand. This work provides strong support for retromutagenesis as a general process creating adaptive mutations during ageing.

Taking higher education to be an arena in which professional and social interaction has a special propensity to overlap, this paper investigates university students’ experiences and perceptions of sexual harassment. Based on survey data, we find varying responses according to their gender and nationality, indicating that men and Danish students are least likely to experience and perceive situations as sexual harassment. Further, we find a wide-spread normalization of certain potentially offensive acts and behaviours. In addition, students report varying degrees of acceptability of certain acts, depending on context. On this basis, we argue that normalization hinders individual students’ ability to recognize and denounce sexual harassment. The influence of social norms on individual experiences and perceptions, we assert, means sexual harassment is neither an objective category nor an individual responsibility. In consequence, issues of sexual harassment can only be dealt with if and when universities assume responsibility for the norms that prevail within their spheres of influence.