Self-pigmenting textiles grown from cellulose-producing bacteria with engineered tyrosinase expression.

Environmental concerns are driving interest in postpetroleum synthetic textiles produced from microbial and fungal sources. Bacterial cellulose (BC) is a promising sustainable leather alternative, on account of its material properties, low infrastructure needs and biodegradability. However, for alternative textiles like BC to be fully sustainable, alternative ways to dye textiles need to be developed alongside alternative production methods. To address this, we genetically engineer Komagataeibacter rhaeticus to create a bacterial strain that grows self-pigmenting BC. Melanin biosynthesis in the bacteria from recombinant tyrosinase expression achieves dark black coloration robust to material use. Melanated BC production can be scaled up for the construction of prototype fashion products, and we illustrate the potential of combining engineered self-pigmentation with tools from synthetic biology, through the optogenetic patterning of gene expression in cellulose-producing bacteria. With this study, we demonstrate that combining genetic engineering with current and future methods of textile biofabrication has the potential to create a new class of textiles.

Taxonomic revision of the native bee subgenus Parasphecodes Smith 1853 in Australia (Hymenoptera: Halictidae: Halictini: Lasioglossum Curtis 1833).

The Australian Lasioglossum Curtis 1833 subgenus Parasphecodes Smith 1853 is revised. Currently, Parasphecodes has 92 named, described species. The monotypic Lasioglossum subgenus Pseudochilalictus Michener 1965 is synonymised with Parasphecodes and its species, L. imitator Michener 1965, is recombined into Parasphecodes. The single known species from New Guinea, L. (Parasphecodes) permetallicum Michener 1965, is included in this revision. Eighteen new species are erected, 69 names are placed into synonymy, 20 new sex associations made and three species currently placed in Parasphecodes are recombined into the Lasioglossum subgenus Ctenonomia Cameron 1903. This revision resolved there are 40 valid species for Parasphecodes. Valid species for Lasioglossum (Parasphecodes) without synonymies are as follows: L. imitator, L. lichatus (Smith 1853), L. loweri (Cockerell 1905), L. olgae (Rayment 1935), L. permetallicum, L. turneri (Cockerell 1914d) and L. waterhousei (Cockerell 1915a). New synonymies proposed for Lasioglossum (Parasphecodes) are as follows: Lasioglossum (Pseudochilalictus Michener 1965) new synonymy = Lasioglossum (Parasphecodes); L. cirriferum (Cockerell 1910) new synonymy, L. insigne (Meyer 1920) new synonymy and L. grande (Meyer 1920) new synonymy = L. altichus (Smith 1853); L. paramelaenum (Cockerell 1922) new synonymy = L. atronitens (Cockerell 1914a); L. bribiense (Cockerell 1916) new synonymy, L. bribiensiforme (Cockerell 1930a) new synonymy, L. butleri (Rayment 1935) new synonymy, L. frenchi (Rayment 1935) new synonymy, L. frenchellum Michener 1965 new synonymy, L. sordidulum (Cockerell 1914c) new synonymy and L. patongensis (Rayment 1948) new synonymy = L. bryotrichum (Cockerell 1912a); L. fumidicaudum (Cockerell 1914b) new synonymy and L. noachinum (Cockerell 1914b) new synonymy = L. carbonarium (Smith 1853); L. cervicale (Cockerell 1915b) new synonymy and L. zamelanum (Cockerell 1930a) new synonymy = L. dissimulator (Cockerell 1914b); L. wilmatae (Cockerell 1929c) new synonymy = L. excultum (Cockerell 1913b); L. arciferum (Cockerell 1914b) new synonymy, L. atrorufescens (Cockerell 1914b) new synonymy, L. fulviventre (Friese 1924) new synonymy, L. leptospermi (Cockerell 1916) new synonymy, L. lichatinum (Cockerell 1922) new synonymy, L. leucorhinum (Cockerell 1926) new synonymy, L. proximum (Rayment 1947) new synonymy, L. testaciventre (Rayment 1953) new synonymy, L. tilachus (Smith 1853) new synonymy, L. tilachiforme (Cockerell 1907) new synonymy, L. tuchilas (Smith 1853) new synonymy, L. anhybodinum (Cockerell 1930a) new synonymy, L. hybodinum (Cockerell 1912a) new synonymy, L. tripunctatum (Cockerell 1929c) new synonymy and L. warburtoni (Cockerell 1906) new synonymy = L. hilactus (Smith 1853); L. frenchi (Cockerell 1904) new synonymy, L. schomburgki (Cockerell 1910) new synonymy, L. speculiferum (Cockerell 1912a) new synonymy, L. sextum (Cockerell 1910) new synonymy, L. solis (Cockerell 1922) new synonymy, L. vermiculatum (Cockerell 1914b) new synonymy and L. vulneratum (Cockerell 1910) new synonymy = L. hiltacus (Smith 1853); L. hirtiventre (Cockerell 1922) new synonymy, L. niveorufum (Friese 1924) new synonymy, L. submeracum (Cockerell 1930a) new synonymy and L. froggatti (Cockerell 1905) new synonymy = L. lacthius (Smith 1853); L. basilautum (Cockerell 1910) new synonymy, L. doddi (Cockerell 1914c) new synonymy, L. paracolletinum (Cockerell 1910) new synonymy, L. pilicolle (Friese 1924) new synonymy, L. scutellatum (Friese 1924) new synonymy and L. vau (Cockerell 1910) new synonymy = L. leichardti (Cockerell 1906); L. annexum (Cockerell 1922) new synonymy, L. latissimum (Cockerell 1915b) new synonymy, L. microdontum (Cockerell 1912a) new synonymy, L. recessum (Cockerell 1914d) new synonymy, L. longmani (Cockerell 1922) new synonymy, L. recantans (Cockerell 1912a) new synonymy and L. rufotegulare (Cockerell 1914e) new synonymy = L. melbournense (Cockerell 1904); L. trimaculatum (Friese 1924) new synonymy = L. musicum (Cockerell 1913a); L. gentianae (Rayment 1951) new synonymy = L. subrussatum (Cockerell 1922); L. fultoni (Cockerell 1914b) new synonymy, L. gibbosum (Friese 1924) new synonymy, L. niveatum (Meyer 1920) new synonymy, L. punctatissimus (Meyer 1920) new synonymy, L. rhodopterum (Cockerell 1914e) new synonymy, L. rubriventre (Friese 1924) new synonymy, L. subfultoni (Cockerell 1930a) new synonymy, L. tepperi (Cockerell 1905) new synonymy, L. notescens (Cockerell 1930a) new synonymy and L. rufulum (Friese 1924) new synonymy = L. sulthica (Smith 1853); L. submoratum (Cockerell 1930a) new synonymy and L. perustum (Cockerell 1914d) new synonymy = L. taluchis (Smith 1853). Eighteen new species are described as follows: L. acristum Walker & Sparks, L. altum Walker & Sparks, L. aspereticulum Walker & Sparks, L. atropum Walker & Sparks, L. bimelasmum Walker & Sparks, L. bipenicillum Walker & Sparks, L. bitrichum Walker & Sparks, L. blyscanatum Walker & Sparks, L. brevipectinatum Walker & Sparks, L. capronum Walker & Sparks, L. ferruginum Walker & Sparks, L. flexosum Walker & Sparks, L. laevidiscum Walker & Sparks, L. recavum Walker & Sparks, L. reticulum Walker & Sparks, L. rutrum Walker & Sparks, L. variegatum Walker & Sparks and L. wcisloi Walker & Sparks. New subgeneric classifications are as follows: L. (Pseudochilalictus) imitator = L. (Parasphecodes) imitator new status, Halictus clarigaster Cockerell 1918 = L. (Ctenonomia) clarigaster new status, Halictus forresti Cockerell 1906 = L. (Ctenonomia) forresti new status, and Halictus tribuarius Rayment 1935 = L. (Ctenonomia) tribuarium new status. These species names, all described by Smith 1853, are anagrams of Halictus. Therefore, they are nouns in apposition and should retain their original species designations as: Lasioglossum (Parasphecodes) altichus (Smith 1853), Lasioglossum (Parasphecodes) hilactus (Smith 1853), Lasioglossum (Parasphecodes) hiltacus (Smith 1853), Lasioglossum (Parasphecodes) lacthius (Smith 1853), Lasioglossum (Parasphecodes) lichatus (Smith 1853), Lasioglossum (Parasphecodes) sulthica (Smith 1853), Lasioglossum (Parasphecodes) talchius (Smith 1853), Lasioglossum (Parasphecodes) taluchis (Smith 1853), Lasioglossum (Parasphecodes) tilachus (Smith 1853) and Lasioglossum (Parasphecodes) tuchilas (Smith 1853). All 40 valid Parasphecodes species, as well as the three species recombined to Ctenonomia, are redescribed. For the Parasphecodes species, keys to both sexes, character groups, taxonomy, citations, species diagnoses, comments, descriptions, scanning electron micrographs, colour montage images, distribution maps, male genitalia and S7S8 line drawings are provided to assist with species identifications.

Adapting a community pharmacy intervention to improve medication safety.

BACKGROUND: Community pharmacies are an ideal location to address challenges of over-the-counter medication safety, yet many successful interventions are only tested in a few pharmacies without expansion, creating unrealized opportunities to improve patient care on a larger scale. Scaling up to numerous pharmacies can be challenging because each community pharmacy has unique needs and layouts and requires individualized adaptation. OBJECTIVES: This paper reports techniques for (a) adapting a community pharmacy intervention to fit the unique physical layout and patient needs of health system pharmacy sites without increasing staff workload, (b) identifying strategies to gather feedback on adaptations from stakeholders, and (c) developing materials to share with pharmacy champions for them to independently implement and sustain the intervention in their organization. PRACTICE DESCRIPTION: The study team collaborated with Aurora Pharmacy, Inc to develop an intervention designed to increase awareness of safe over-the-counter medication use for older adults. PRACTICE INNOVATION: Senior Safe, a community pharmacy-based intervention, was designed, implemented, and tested using the Exploration, Preparation, Implementation, and Sustainment implementation framework. EVALUATION METHODS: Senior Safe was adapted through pilot testing and a randomized control trial. Feedback was collected from key stakeholders, including pharmacy staff, older adults, and a research advisory group. RESULTS: A finalized version of Senior Safe, as well as an implementation package, was provided to Aurora Pharmacy to integrate into all 63 sites. CONCLUSION: This multiphase study illustrated that refining an intervention is possible and welcomed by pharmacy staff, but it requires time, resources, and funds to create an impactful, sustainable community pharmacy intervention.

Fostering greater recognition of Caribbean traditional plant knowledge.

The Caribbean is a hotspot of biological and cultural diversity, manifested in traditional plant knowledge of Afrodescendant peoples and other ethnicities. To strengthen the visibility of this knowledge in research, education, and policy making, we propose an eight-step action plan centered on reciprocal relationships with Caribbean plant stewards, especially subsistence farmers.

Development of in silico models to predict viscosity and mouse clearance using a comprehensive analytical data set collected on 83 scaffold-consistent monoclonal antibodies.

Biologic drug discovery pipelines are designed to deliver protein therapeutics that have exquisite functional potency and selectivity while also manifesting biophysical characteristics suitable for manufacturing, storage, and convenient administration to patients. The ability to use computational methods to predict biophysical properties from protein sequence, potentially in combination with high throughput assays, could decrease timelines and increase the success rates for therapeutic developability engineering by eliminating lengthy and expensive cycles of recombinant protein production and testing. To support development of high-quality predictive models for antibody developability, we designed a sequence-diverse panel of 83 effector functionless IgG1 antibodies displaying a range of biophysical properties, produced and formulated each protein under standard platform conditions, and collected a comprehensive package of analytical data, including in vitro assays and in vivo mouse pharmacokinetics. We used this robust training data set to build machine learning classifier models that can predict complex protein behavior from these data and features derived from predicted and/or experimental structures. Our models predict with 87% accuracy whether viscosity at 150 mg/mL is above or below a threshold of 15 centipoise (cP) and with 75% accuracy whether the area under the plasma drug concentration-time curve (AUC0-672 h) in normal mouse is above or below a threshold of 3.9 × 106 h x ng/mL.

Integrating simulation into surgical training: a qualitative case study of a national programme.

BACKGROUND: Applying simulation-based education (SBE) into surgical curricula is challenging and exacerbated by the absence of guidance on implementation processes. Empirical studies evaluating implementation of SBE interventions focus primarily on outcomes. However, understanding the processes involved in organising, planning, and delivering SBE adds knowledge on how best to develop, implement, and sustain surgical SBE. This study used a reform of early years surgical training to explore the implementation of a new SBE programme in Scotland. It aimed to understand the processes that are involved in the relative success (or failure) when implementing surgical SBE interventions. METHODS: This qualitative case study, underpinned by social constructionism, used publicly available documents and the relevant surgical SBE literature to inform the research focus and contextualise data obtained from semi-structured interviews with core surgical trainees (n = 46), consultant surgeons (n = 25), and key leaders with roles in surgical training governance in Scotland (n = 7). Initial data coding and analysis were inductive. Secondary data analysis was then undertaken using Normalisation Process Theory (NPT). NPTs' four constructs (coherence, cognitive participation, collective action, reflexive monitoring) provided an explanatory framework for scrutinising how interventions are implemented, embedded, and integrated into practice, i.e. the "normalisation" process. RESULTS: Distributed leadership (individual SBE initiatives assigned to faculty but overall programme overseen by a single leader) and the quality improvement practise of iterative refinement were identified as key novel processes promoting successful normalisation of the new SBE programme. Other processes widely described in the literature were also identified: stakeholder collaboration, personal contacts/relational processes, effective communication, faculty development, effective leadership, and tight programme management. The study also identified that learners valued SBE activities in group- or team-based social environments over isolated deliberate practice. CONCLUSIONS: SBE is most effective when designed as a comprehensive programme aligned to the curriculum. Programmes incorporating both group-based and isolated SBE activities promote deliberate practice. Distributed leadership amongst faculty attracts wide engagement integral to SBE programme implementation, while iterative programme refinement through regular evaluation and action on feedback encourages integration into practice. The knowledge contributed by critically analysing SBE programme implementation processes can support development of much needed guidance in this area.

Context matters in curriculum reform: An analysis of change in surgical training.

INTRODUCTION: Education and training reforms are typically devised by accreditation bodies and rolled out nationally. This top-down approach is positioned as contextually independent, yet context is highly influential in shaping the impact of change. Given this, it is critical to consider how curriculum reform plays out as it meets local settings. We have therefore used a national-level curriculum reform process of surgical training, Improving Surgical Training (IST), to examine the influence of context in IST implementation across two UK countries. METHODS: Adopting a case study approach, we used document data for contextualisation purposes and semi-structured interviews with key stakeholders across multiple organisations (n = 17, plus four follow-up interviews) as our main source of data. Initial data coding and analysis were inductive. We followed this with a secondary analysis using Engeström's second-generation activity theory nested within an overarching framework of complexity theory to help tease out some key elements of IST development and implementation. RESULTS: The introduction of IST into the surgical training system was historically situated within a landscape of previous reforms. IST's aims collided with existing practices and rules, thus creating tensions. In one country, the systems of IST and surgical training came together to some extent, mostly due to processes of social networks, negotiation and leverage nested in a relatively cohesive setting. These processes were not apparent in the other country, and instead of transformative change, the system contracted. Change was not integrated, and the reform was halted. CONCLUSIONS: Our use of a case study approach and complexity theory deepens understanding of how history, systems and contexts interact to facilitate or inhibit change within one area of medical education. Our study paves the way for further empirical work examining the influence of context in curriculum reform, and thus determining how best to bring about change in practice.

Scotland's "Incentivised Laparoscopy Practice" programme: Engaging trainees with take-home laparoscopy simulation.

BACKGROUND: The transfer validity of portable laparoscopy simulation is well established. However, attempts to integrate take-home simulation into surgical training have met with inconsistent engagement worldwide, as for example in our 2014-15 study of an Incentivised Laparoscopy Practice programme (ILPv1). Drawing on learning from our subsequent multi-centre study examining barriers and facilitators, we revised the programme for 2018 onwards. We now report on engagement with the 2018-2022 versions of this home-based simulation programme (ILP v2.1-2.3). METHODS: In ILP v2.1-2.3, three consecutive year-groups of new-start Core Surgical Trainees (n = 48, 46 and 53) were loaned portable simulators. The 6-month education programme included induction, technical support, and intermittent feedback. Six tasks were prescribed, with video instruction and charting of metric scores. Video uploads were required and scored by faculty. A pass resulted in an eCertificate, expected at Annual Review (but not mandatory for progression). ILP was set within a wider reform, "Improving Surgical Training". RESULTS: ILP v2.1-2.3 saw pass rates of 94%, 76% and 70% respectively (45/48, 35/46 and 37/53 trainees), compared with only 26% (7/27) in ILP v1, despite now including some trainees not intending careers in laparoscopic specialties. The ILP v2.2 group all reported their engagement with the whole simulation strategy was hampered by the COVID19 pandemic. CONCLUSIONS: Simply providing take-home simulators, no matter how good, is not enough. To achieve trainee engagement, a whole programme is required, with motivated learners, individual and group practice, intermittent feedback, and clear goals and assessments. ILP is a complex intervention, best understood as a "reform within a reform, within a context."

Scratching beneath the surface: How organisational culture influences curricular reform.

INTRODUCTION: Curricular reform is often proposed as the means to improve medical education and training. However, reform itself may not lead to noticeable change, possibly because the influence of organisational culture on change is given insufficient attention. We used a national reform of early-years surgical training as a natural opportunity to examine the interplay between organisational culture and change in surgical education. Our specific research question was: in what ways did organisational culture influence the implementation of Improving Surgical Training (IST)? METHODS: This is a qualitative study underpinned by social constructivism. Interviews were conducted with core surgical trainees (n = 46) and their supervising consultants (n = 25) across Scotland in 2020-2021. Data coding and analysis were initially inductive. The themes indicated the importance of many cultural factors as barriers or enablers to IST implementation. We therefore carried out a deductive, secondary data analysis using Johnson's (1988) cultural web model to identify and examine the different elements of organisational culture and their impact on IST. RESULTS: The cultural web enabled a detailed understanding of how organisational culture influenced IST implementation as per Johnson's six elements-Rituals and Routines (e.g. departmental rotas), Stories (e.g. historical training norms and culture), Symbols (e.g. feedback mechanisms, visibility and value placed on education), Power Structures (e.g. who has the power in local contexts), Organisational Structures (e.g. relationships and accountability) and the Control System (e.g. consultant job plans and service targets)-and how these interact. However, it did not shed light on the influence of exogenous events on change. CONCLUSION: Our data reveal cultural reasons why this curricular reform met with varying degrees of success across different hospital sites, reinforcing that curricular reform is not simply about putting recommendations into practice. Many different aspects of context must be considered when planning and evaluating change in medical education and training.

"It's making me think outside the box at times": a qualitative study of dynamic capabilities in surgical training.

Craft specialties such as surgery endured widespread disruption to postgraduate education and training during the pandemic. Despite the expansive literature on rapid adaptations and innovations, generalisability of these descriptions is limited by scarce use of theory-driven methods. In this research, we explored UK surgical trainees' (n = 46) and consultant surgeons' (trainers, n = 25) perceptions of how learning in clinical environments changed during a time of extreme uncertainty (2020/2021). Our ultimate goal was to identify new ideas that could shape post-pandemic surgical training. We conducted semi-structured virtual interviews with participants from a range of working/training environments across thirteen Health Boards in Scotland. Initial analysis of interview transcripts was inductive. Dynamic capabilities theory (how effectively an organisation uses its resources to respond to environmental changes) and its micro-foundations (sensing, seizing, reconfiguring) were used for subsequent theory-driven analysis. Findings demonstrate that surgical training responded dynamically and adapted to external and internal environmental uncertainty. Sensing threats and opportunities in the clinical environment prompted trainers' institutions to seize new ways of working. Learners gained from reconfigured training opportunities (e.g., splitting operative cases between trainees), pan-surgical working (e.g., broader surgical exposure), redeployment (e.g., to medical specialties), collaborative working (working with new colleagues and in new ways) and supervision (shifting to online supervision). Our data foreground the human resource and structural reconfigurations, and technological innovations that effectively maintained surgical training during the pandemic, albeit in different ways. These adaptations and innovations could provide the foundations for enhancing surgical education and training in the post-pandemic era.

Natural source zone depletion (NSZD) insights from over 15 years of research and measurements: A multi-site study.

Site-average Natural Source Zone Depletion (NSZD) rates measured from 40 petroleum light non-aqueous phase liquid (LNAPL) source zone sites were compiled from researchers, project reports, and scientific papers. At each site, the following data were compiled: i) general site location; ii) LNAPL fuel type; iii) measurement method, number of locations, and number of measurements per location; and iv) calculated site-average NSZD rate in liters per hectare per year (L/ha/yr) per site and the associated measurement method (i.e., Gradient Method, Carbon Traps, Dynamic Closed Chamber (DCC), or Thermal Monitoring). The resulting dataset showed site-average NSZD rates that ranged from 650 to 152,000 L/ha/yr (70 to 16,250 gallons per acre per year (gal/acre/yr)), with a median value of 9,540 L/ha/yr (1,020 gal/acre/yr). The median site-average NSZD rate by type of fuel spill did not show a statistically significant difference between fuel types. When comparing the different NSZD measurement methods applied to the same sites, the site-average NSZD rates differed by up to 4.8 times (i.e., ratio of faster rate to slower rate), with a median difference of 2.1 times. No clear bias was observed between NSZD rate measurement methods. At four sites with calculations of NSZD rates by season, NSZD rates were typically higher during summer and fall compared to winter and spring. For these sites, Q10 values (a measure of the increase in NSZD rate associated with a 10 °C increase in temperature) ranged from 0.8 to 15.1, with a median of 2.2. The implications of this study suggest that increasing mean annual soil temperature at a site using engineered methods could potentially increase the biodegradation rate (e.g., an increase of 10 °C could double the NSZD rate). Finally, for five sites with site-average NSZD rates for multiple years, average NSZD rates varied by 1.1 to 4.9 times across years. Overall, the evaluation of NSZD rates measured at 40 LNAPL sites suggests that measurable NSZD occurs across a broad range of LNAPL sites. Although NSZD rates vary across sites, fuel type is not the primary factor explaining observed differences in rates.

Consenso chileno de diagnóstico y manejo de neoplasias quísticas pancreáticas. Club de páncreas Chile-SCHGE / Chilean consensus on diagnosis and management of pancreatic cystic neoplasms

Pancreatic cystic neoplasms (PCN) are frequently detected on abdominal images performed for non-pancreatic indications. Their prevalence in asymptomatic population ranges from 2.7 to 24.8%, and increases with age. There are several types of pancreatic cysts. Some may contain cancer or have malignant potential, such as mucinous cystic neoplasms, including mucinous cystadenoma (MCN) and intraductal papillary mucinous neoplasms (IPMN). In contrast, others are benign, such as serous cystadenoma (SCA). However, even those cysts with malignant potential rarely progress to cancer. Currently, the only treatment for pancreatic cysts is surgery, which is associated with high morbidity and occasional mortality. The Board of the Chilean Pancreas Club of the Chilean Gastroenterology Society developed the first Chilean multidisciplinary consensus for diagnosis, management, and surveillance of PCN. Thirty experts were invited and answered 21 statements with five possible alternatives: 1) fully agree; 2) partially agree; 3) undecided; 4) disagree and 5) strongly disagree. A consensus was adopted when at least 80% of the sum of the answers "fully agree" and "partially agree" was reached. The consensus was approved by the Board of Directors of the Chilean Pancreas Club for publication.

Komagataeibacter Tool Kit (KTK): A Modular Cloning System for Multigene Constructs and Programmed Protein Secretion from Cellulose Producing Bacteria.

Bacteria proficient at producing cellulose are an attractive synthetic biology host for the emerging field of Engineered Living Materials (ELMs). Species from the Komagataeibacter genus produce high yields of pure cellulose materials in a short time with minimal resources, and pioneering work has shown that genetic engineering in these strains is possible and can be used to modify the material and its production. To accelerate synthetic biology progress in these bacteria, we introduce here the Komagataeibacter tool kit (KTK), a standardized modular cloning system based on Golden Gate DNA assembly that allows DNA parts to be combined to build complex multigene constructs expressed in bacteria from plasmids. Working in Komagataeibacter rhaeticus, we describe basic parts for this system, including promoters, fusion tags, and reporter proteins, before showcasing how the assembly system enables more complex designs. Specifically, we use KTK cloning to reformat the Escherichia coli curli amyloid fiber system for functional expression in K. rhaeticus, and go on to modify it as a system for programming protein secretion from the cellulose producing bacteria. With this toolkit, we aim to accelerate modular synthetic biology in these bacteria, and enable more rapid progress in the emerging ELMs community.

Bacterial cellulose spheroids as building blocks for 3D and patterned living materials and for regeneration.

Engineered living materials (ELMs) based on bacterial cellulose (BC) offer a promising avenue for cheap-to-produce materials that can be programmed with genetically encoded functionalities. Here we explore how ELMs can be fabricated in a modular fashion from millimetre-scale biofilm spheroids grown from shaking cultures of Komagataeibacter rhaeticus. Here we define a reproducible protocol to produce BC spheroids with the high yield bacterial cellulose producer K. rhaeticus and demonstrate for the first time their potential for their use as building blocks to grow ELMs in 3D shapes. Using genetically engineered K. rhaeticus, we produce functionalized BC spheroids and use these to make and grow patterned BC-based ELMs that signal within a material and can sense and report on chemical inputs. We also investigate the use of BC spheroids as a method to regenerate damaged BC materials and as a way to fuse together smaller material sections of cellulose and synthetic materials into a larger piece. This work improves our understanding of BC spheroid formation and showcases their great potential for fabricating, patterning and repairing ELMs based on the promising biomaterial of bacterial cellulose.

Transthyretin-mediated protein and peptide oligomerization for enhanced target clustering.

Advances in cancer research have led to the development of new therapeutics with significant and durable responses such as immune checkpoint inhibitors. More recent therapies aim to stimulate anti-tumor immune responses by targeting the tumor necrosis factor (TNF) receptors, however this approach has been shown to require clustering of receptors in order to achieve a significant response. Here we present a perspective on using transthyretin, a naturally occurring serum protein, as a drug delivery platform to enable cross-linking independent clustering of targets. TTR forms a stable homo-tetramer with exposed termini that make TTR a highly versatile platform for generating multimeric antibody fusions to enable enhanced target clustering. Fusions with antibodies or Fabs targeting TRAILR2 were shown to have robust cytotoxic activity in vitro and in vivo in colorectal xenograft models demonstrating that TTR is a highly versatile, stable, therapeutic fusion platform that can be used with antibodies, Fabs and other bioactive fusion partners and has broad applications in oncology and infectious disease research.

Simulation for technical skill acquisition in open vascular surgery.

BACKGROUND: Simulation has an increasingly prominent role in modern vascular surgery training. However, it is important to understand how simulation is most effectively delivered to best use the time and resources available. The aim of this narrative review is therefore to critically appraise open technical skill acquisition in the operating room environment and provide recommendations for the future development of evidence-based simulation for open vascular surgery. METHODS: A systematic search strategy was used to retrieve relevant studies from PubMed, Medline, Web of Science, EMBASE, and the Cochrane databases in July 2019. Included papers were independently screened by two reviewers. Data were subsequently extracted using a standardized proforma and thematically analyzed. RESULTS: Thirteen studies were included. All demonstrated that simulation is effective in improving confidence and/or competence in performing open technical skills when assessed by previously validated metrics. However, not all participants or course schedules achieved equal benefit, with distributed practice for junior trainees over several weeks achieving a greater improvement in technical skill compared with senior trainees or longer course schedules for some tasks. CONCLUSIONS: Simulation can be an effective adjunct to traditional operative experience for technical skill acquisition in open vascular surgery. Future work should focus on developing models to address a wider range of training needs, as well as further defining the optimum schedule for the style, content, and timing of simulation for specific learner groups.

Moving forward through the in silico modeling of tuberculosis: a further step with UISS-TB.

BACKGROUND: In 2018, about 10 million people were found infected by tuberculosis, with approximately 1.2 million deaths worldwide. Despite these numbers have been relatively stable in recent years, tuberculosis is still considered one of the top 10 deadliest diseases worldwide. Over the years, Mycobacterium tuberculosis has developed a form of resistance to first-line tuberculosis treatments, specifically to isoniazid, leading to multi-drug-resistant tuberculosis. In this context, the EU and Indian DBT funded project STriTuVaD-In Silico Trial for Tuberculosis Vaccine Development-is supporting the identification of new interventional strategies against tuberculosis thanks to the use of Universal Immune System Simulator (UISS), a computational framework capable of predicting the immunity induced by specific drugs such as therapeutic vaccines and antibiotics. RESULTS: Here, we present how UISS accurately simulates tuberculosis dynamics and its interaction within the immune system, and how it predicts the efficacy of the combined action of isoniazid and RUTI vaccine in a specific digital population cohort. Specifically, we simulated two groups of 100 digital patients. The first group was treated with isoniazid only, while the second one was treated with the combination of RUTI vaccine and isoniazid, according to the dosage strategy described in the clinical trial design. UISS-TB shows to be in good agreement with clinical trial results suggesting that RUTI vaccine may favor a partial recover of infected lung tissue. CONCLUSIONS: In silico trials innovations represent a powerful pipeline for the prediction of the effects of specific therapeutic strategies and related clinical outcomes. Here, we present a further step in UISS framework implementation. Specifically, we found that the simulated mechanism of action of RUTI and INH are in good alignment with the results coming from past clinical phase IIa trials.

Engineering Bacterial Cellulose by Synthetic Biology.

Synthetic biology is an advanced form of genetic manipulation that applies the principles of modularity and engineering design to reprogram cells by changing their DNA. Over the last decade, synthetic biology has begun to be applied to bacteria that naturally produce biomaterials, in order to boost material production, change material properties and to add new functionalities to the resulting material. Recent work has used synthetic biology to engineer several Komagataeibacter strains; bacteria that naturally secrete large amounts of the versatile and promising material bacterial cellulose (BC). In this review, we summarize how genetic engineering, metabolic engineering and now synthetic biology have been used in Komagataeibacter strains to alter BC, improve its production and begin to add new functionalities into this easy-to-grow material. As well as describing the milestone advances, we also look forward to what will come next from engineering bacterial cellulose by synthetic biology.