We can do it - Empowering learning.

We all need to adapt constantly in order to thrive in our ever-changing complex world. Thus, education and educators need to empower learners to develop a mindset and skills set of lifelong and lifewide learning. Papers from around the globe, reflecting the current drivers in education, were published in the FEMS Microbiology Letters virtual Thematic Issue 'Empowering learning' in October 2023. Its content is reviewed here to provide the context for further discussions within the professional community. The focus is on being inclusive, on lowering and removing barriers to learning, and fostering a sense of belonging. So learning communities can be established and connected in support of learning with and from each other for societal benefit, and this is discussed. Authentic learning approaches are highlighted with respect to the inherent opportunities for learners to take ownership for their learning, yet learn collaboratively, and develop resilience over time. Supporting learners in becoming self-regulated and realizing their full potential is truly worthwhile.

Absence makes the mind grow stronger - Educating in a pandemic and beyond.

With more than one academic year into the pandemic, it is timely to consider the lessons we learnt, and how they could shape education in the future. Papers from around the globe, reflecting on the directions we took and could take, were published in the FEMS Microbiology Letters virtual Thematic Issue 'Educating in a pandemic and beyond' in October 2021. Its content is reviewed here to facilitate discussions within the professional community. Online platforms and tools, that allowed a rapid emergency response, are covered, as well as enhancing student engagement, complementing and blending in-person activities with online elements for more flexible and accessible learning opportunities, the need for educator training, and improving science literacy overall and microbiology literacy specifically. As we go forward, in order to benefit from blended and flexible learning, we need to select our approaches based on evidence, and mindful of the potential impact on learners and educators. Education did not only continue during the pandemic, but it evolved, leading us into the future.

Be prepared - Learning for the future.

Forward-looking papers from around the globe, addressing themes of current educational practice, were published in the FEMS Microbiology Letters virtual Thematic Issue 'Learning for the future' in October 2019. Its dynamic content is reviewed here to facilitate discussions within the professional community. The focus is on preparing generations of 21st century professionals as lifelong learners for yet unknown working environments and demands. Active and authentic learning is covered in various contexts from interactive lectures, via innovative tasks exploring the scientific method to conducting research over a distance. Naturally, authentic assessment features. Moreover, authentic learning is taken out of the classroom into the community as service learning for students to develop skills and see unexpected Science, Technology, Engineering and Maths career opportunities. Approaches to public engagement with science are presented including through art. Art is also brought into the classroom, as well as educational games, to foster creativity. Additionally, sharing science with large audiences on social media is discussed. Future-proofing education requires being innovative and imaginative. This comes with challenges and risks, but being prepared is worth it.

Keeping education fresh-not just in microbiology.

Innovative practice from around the globe, addressing a range of recent educational themes and trends, was published in the FEMS Microbiology Letters virtual Thematic Issue 'Keeping Education Fresh' in October 2017. Its thought-provoking content is reviewed here to more directly facilitate reflections and discussions in the professional community. The focus is on best practice approaches when enhancing student engagement, how to adjust those to the diversity of learners, learning situations and infrastructures, and to a broad range of subjects. The need for authentic learning and to move away from didactic teaching is emphasized. The 'students as researchers' theme is featured e.g. in context of service learning. Creative approaches are presented such as using performing arts, popular culture and gamification. The development of interdisciplinary and intercultural competences, and the exploration of socioscientific themes and philosophical issues are considered. Revisions of curricula and programmes, reflective of educational advancements and sector drivers, are discussed from undergraduate to postgraduate and professional specialist level also in light of problem-based learning, interactive distance and on-campus learning, and even the legacy of Massive Open Online Courses. Such changes always require resources and skills, and carry risks. Yet, innovation is a risk worth taking to keep education fresh.

Edging into the future: education in microbiology and beyond.

State-of-the-art papers from around the globe addressing current topics in education were published in the FEMS Microbiology Letters virtual Thematic Issue 'Education' in November 2015 (http://femsle.oxfordjournals.org/content/thematic-issue-education), which was innovative and well received by microbiologists and other educators. Its unique content is reviewed here to facilitate broader access and further discussions in the professional community. Best practice in supporting school teaching and exposing students to concepts from other disciplines is presented in context of inspiring the next generations, where also historical microbiology can be drawn upon. Technology-enhanced education is discussed including its applications (e.g. lecture podcasts for flipped learning, learning from experts via videoconference). Authentic learning is covered with examples of research-led teaching, water and showerhead biofilm analyses and participation in the International Genetically Engineered Machines competition. Enhancing employability is focussed on, including supporting personal development and work-readiness in general and for the changing nature of the microbiology profession. International mobility develops international awareness but challenges teachers. Teaching training, teaching excellence and dissemination of best practice are reviewed. Times of challenge and change in the Higher Education landscape motivate us to improve educational approaches and frameworks, so that we are prepared for new topics to emerge as current topics in education.

Teaching matters-academic professional development in the early 21st century.

Academic work at different career stages has changed and a broadened portfolio of expertise enables academics to adapt, maintain and advance their career. Development related to research activity is naturally driven by methodology and technology. Institutions and peers largely support development in the contexts of dissemination, measuring impact and obtaining funding. A European Commission High Level Group recommended pedagogic training for everyone teaching in Higher Education by 2020 with mandatory continuing professional development and with academic staff recruitment and promotion being linked to teaching performance. Early career teaching experience is already an expectation, and advantage is gained by developing recognized teaching expertise. More senior academics gain an advantage through recognition of higher levels of expertise, also covering elements of leadership and innovation in teaching. This review aims to raise awareness particularly of teaching-related skills within the dimensions of academic professional development in Higher Education, outlining some general directions for development and recognition in context of current challenges to support planning and identifying training needs and opportunities at different career stages.

On your marks, get set, go!-lessons from the UK in enhancing employability of graduates and postgraduates.

Employers expect graduates and postgraduates to demonstrate their education through more than good grades. Learning activities that develop subject skills during formalized programmes of undergraduate and postgraduate study also develop employability skills, if the curriculum is suitably aligned, and developmental planning is supported. Only little extra provision is required, but all development needs to be explicitly signposted to the learner, and the curriculum should be developed in consultation with employers. This review aims to raise awareness of current issues in the context of enhancing employability that arise from an increased global competition on the job market and the expectation of the Higher Education sector to produce work-ready graduates and postgraduates that are well equipped to adapt to a quickly changing work environment particularly due to transferable skills. In the context of lessons from the UK, these current issues and employability are discussed, and approaches to Personal Development Planning that prepare students for lifelong learning and that enable communicating and evidencing achievement are addressed. Issues specific to postgraduates, how actual work experience should be maximized as well as other career influences such as learned societies and social networking are highlighted.

Using folding promoting agents in recombinant protein production: a review.

Recombinant production has become an invaluable tool for supplying research and therapy with proteins of interest. The target proteins are not in every case soluble and/or correctly folded. That is why different production parameters such as host, cultivation conditions and co-expression of chaperones and foldases are applied in order to yield functional recombinant protein. There has been a constant increase and success in the use of folding promoting agents in recombinant protein production. Recent cases are reviewed and discussed in this chapter. Any impact of such strategies cannot be predicted and has to be analyzed and optimized for the corresponding target protein. The in vivo effects of the agents are at least partially comparable to their in vitro mode of action and have been studied by means of modern systems approaches and even in combination with folding/activity screening assays. Resulting data can be used directly for experimental planning or can be fed into knowledge-based modelling. An overview of such technologies is included in the chapter in order to facilitate a decision about the potential in vivo use of folding promoting agents.

RNA-based sandwich hybridisation method for detection of lactic acid bacteria in brewery samples.

Recently we showed the applicability and sensitivity of the RNA-based sandwich hybridisation assay (SHA) for detection of gram-negative cells in environmental samples [Leskelä, T., Tilsala-Timisjärvi, A., Kusnetsov, J., Neubauer, P., Breitenstein, A., 2005. Sensitive genus-specific detection of Legionella by a 16S rRNA based sandwich hybridization assay. J. Microbiol. Met. 62, 167-179.]. In this study the aim was to test and optimise this method for the detection of gram-positive cells from brewery yeast slurries that contain up to 10(9) yeast cells/ml. Eleven new oligonucleotide probes were designed for group-specific detection of different beer-spoiling lactic acid bacteria of the genera Lactobacillus and Pediococcus. Functionality of the designed probes was shown by testing individual and paired probes using in vitro transcribed 16S rRNA and crude cell extracts as samples. Various simple and fast cell disruption methods were evaluated for the efficient disruption of lactobacilli and pediococci. The applicability of the designed oligonucleotide probes and the SHA for detection of brewery contaminants was demonstrated using both artificial and actual yeast slurry samples from brewery fermentation tanks with either fluorimetric readout or an electric biochip analyser.

Murine Wnt-1 with an internal c-myc tag recombinantly produced in Escherichia coli can induce intracellular signaling of the canonical Wnt pathway in eukaryotic cells.

Wnt-1 belongs to the Wnt family of secreted glycoproteins inducing an intracellular signaling pathway involved in cell proliferation, differentiation, and pattern formation. The canonical branch is one of three known branches. This is also valid in vitro, and Wnts can be considered beneficial for culturing primary cells from organs, provided Wnts are available and applicable even with cells of different species. It was shown here that internally c-myc-tagged murine Wnt-1 produced in the heterologous host Escherichia coli was appropriate for inducing intracellular signaling of the canonical Wnt pathway in eukaryotic cells via stabilization of cytosolic beta-catenin. The pioneering injection of the protein into the blastocoels of Xenopus laevis embryos led to axis duplication and suppression of head formation. Applying the recombinant murine Wnt-1 to metanephric mesenchyme activated the tubulogenic program. The signal-inducing activity of the recombinant protein was also positively demonstrated in the TOP-flash reporter assay. Although Wnts were purified recently from the growth media of stably transfected eukaryotic cell lines, the production of active Wnt proteins in pro- or eukaryotic microorganisms reportedly has never been successful. Here soluble production in E. coli and translocation into the oxidizing environment of the periplasm were achieved. The protein was purified using the internal c-myc tag. The effect on the eukaryotic cells implies that activity was retained. Thus, this approach could make recombinant murine Wnt-1 available as a good starting point for other Wnts needed, for example, for maintaining and differentiating stem cells, organ restoration therapy, and tissue engineering.

Folding-promoting agents in recombinant protein production.

Recombinant protein production has become an essential tool for providing the necessary amounts of a protein of interest to either research or therapy. The target proteins are not in every case soluble and/or correctly folded. That is why different production parameters, such as host, cultivation conditions, and co-expression of chaperones and foldases, are applied in order to gain functional recombinant proteins. Furthermore, the addition of folding-promoting agents during the cultivation is increasingly performed. The impact of all these strategies cannot be predicted and must be analyzed and optimized for the corresponding target protein. In this chapter recent cases of using folding-promoting agents in recombinant protein production are reviewed and discussed with respect to their in vivo applicability. Their effects in the cells are mostly not known in detail but at least partially comparable with the in vitro mode of action. The corresponding in vitro effects are also included in the chapter in order to facilitate a decision about their potential in vivo use.

Inclusion bodies: formation and utilisation.

The efficient in vivo folding of many heterologous proteins is a major bottleneck of high level production in bacterial hosts and simple optimisation protocols have not been available yet. Therefore, inclusion body (IB) based processes play a major role as a potential strategy for the production of complex recombinant proteins. These processes combine the advantages of a high accumulation of the target protein in well-characterised bacteria such as Escherichia coli, efficient strategies for IB isolation, purification and in vitro protein refolding without the need of complicated coexpression systems. Recent advances in the molecular physiology of IB formation and resolubilisation allow straight-forward optimisation of fermentation processes to obtain a high-quality product. In addition, simple strategies have been developed to optimise the purification and renaturation of disulfide bond containing proteins making a fast transfer of such processes into the industrial production scale realistic.

Novel bacterial membrane surface display system using cell wall-less L-forms of Proteus mirabilis and Escherichia coli.

We describe a novel membrane surface display system that allows the anchoring of foreign proteins in the cytoplasmic membrane (CM) of stable, cell wall-less L-form cells of Escherichia coli and Proteus mirabilis. The reporter protein, staphylokinase (Sak), was fused to transmembrane domains of integral membrane proteins from E. coli (lactose permease LacY, preprotein translocase SecY) and P. mirabilis (curved cell morphology protein CcmA). Both L-form strains overexpressed fusion proteins in amounts of 1 to 100 microg ml(-1), with higher expression for those with homologous anchor motifs. Various experimental approaches, e.g., cell fractionation, Percoll gradient purification, and solubilization of the CM, demonstrated that the fusion proteins are tightly bound to the CM and do not form aggregates. Trypsin digestion, as well as electron microscopy of immunogold-labeled replicas, confirmed that the protein was localized on the outside surface. The displayed Sak showed functional activity, indicating correct folding. This membrane surface display system features endotoxin-poor organisms and can provide a novel platform for numerous applications.