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The world of audio is undergoing seismic changes. Traditionally a space dominated by linear radio programmes, in the 2020s this form of listening remains relevant but is now being challenged by other forms and global media platforms. In particular, podcasting offers producers opportunities to pitch for commissions from brands and platforms or to make work independently. \ud Historically, podcasting was a medium led and shaped by amateurs and distributed for free by independent creators. In the 2020s this relationship began to shift. In the United Kingdom, audio producers are now finding success outside of radio, in what was once a market dominated by the BBC. Forming what we might term a 'new audio economy', producers and creatives are working across multiple forms of production in the long tail of audio media\ud Through interviews and analysis, this article will explore the impact of podcasting and other forms of audio production on the UK Independent radio/audio sector, noting the influence of the BBC and shifting patterns of production within the sector\ud Keywords: Audio Production, Podcasting, Radio, BBC

Several new structures of three types of protein complexes, obtained by cryo-electron microscopy (cryo-EM) and published between 2019 and 2021, identify a new family of natural molecular wheels, the "5:2 rotary motors." These span the cytoplasmic membranes of bacteria, and their rotation is driven by ion flow into the cell. They consist of a pentameric wheel encircling a dimeric axle within the cytoplasmic membrane of both Gram-positive and gram-negative bacteria. The axles extend into the periplasm, and the wheels extend into the cytoplasm. Rotation of these wheels has never been observed directly; it is inferred from the symmetry of the complexes and from the roles they play within the larger systems that they are known to power. In particular, the new structure of the stator complex of the Bacterial Flagellar Motor, MotA(5)B(2), is consistent with a "wheels within wheels" model of the motor. Other 5:2 rotary motors are believed to share the core rotary function and mechanism, driven by ion-motive force at the cytoplasmic membrane. Their structures diverge in their periplasmic and cytoplasmic parts, reflecting the variety of roles that they perform. This review focuses on the structures of 5:2 rotary motors and their proposed mechanisms and functions. We also discuss molecular rotation in general and its relation to the rotational symmetry of molecular complexes., Frontiers in Microbiology, 13, ISSN:1664-302X

This document summarizes suggestions of the central sleep apnea (CSA) Technical Expert Panel working group. This paper shares our vision for bringing the right device to the right patient at the right time. For patients with CSA, current coverage criteria do not align with guideline treatment recommendations. For example, CPAP and oxygen therapy are recommended but not covered for CSA. On the other hand, bilevel positive airway pressure (BPAP) without a backup rate may be a covered therapy for OSA, but it may worsen CSA. Narrow coverage criteria that require near elimination of obstructive breathing events on CPAP or BPAP in the spontaneous mode, even if at poorly tolerated pressure levels, may preclude therapy with BPAP with backup rate or adaptive servoventilation, even when those devices provide demonstrably better therapy. CSA is a dynamic disorder that may require different treatments over time, sometimes switching from one device to another; an example is switching from BPAP with backup rate to an adaptive servoventilation with automatic end-expiratory pressure adjustments, which may not be covered. To address these challenges, we suggest several changes to the coverage determinations, including: (1) a single simplified initial and continuing coverage definition of CSA that aligns with OSA; (2) removal of hypoventilation terminology from coverage criteria for CSA; (3) all effective therapies for CSA should be covered, including oxygen and all PAP devices with or without backup rates or servo-mechanisms; and (4) patients shown to have a suboptimal response to one PAP device should be allowed to add oxygen or change to another PAP device with different capabilities if shown to be effective with testing.

The generation of reliable mAb of unique and desired specificities serves as a valuable technology to study protein expression and function. However, standard approaches to mAb generation usually involve large-scale protein purification and intensive screening. In this study, we describe an optimized high-throughput proof-of-principle method for the expanded generation, enrichment, and screening of mouse hybridomas secreting mAb specific for a protein of interest. Briefly, we demonstrate that small amounts of a biotinylated protein of interest can be used to generate tetramers for use as prime-boost immunogens, followed by selective enrichment of Ag-specific B cells by magnetic sorting using the same tetramers prior to hybridoma generation. This serves two purposes: 1) to effectively expand both low- and high-affinity B cells specific for the antigenic bait during immunization and 2) to minimize subsequent laborious hybridoma efforts by positive selection of Ag-specific, Ab-secreting cells prior to hybridoma fusion and validation screening. Finally, we employ a rapid and inexpensive screening technology, CELLISA, a high-throughput validation method that uses a chimeric Ag fused to the CD3ζ signaling domain expressed on enzyme-generating reporter cells; these reporters can detect specific mAb in hybridoma supernatants via plate-bound Ab-capture arrays, thereby easing screening. Using this strategy, we generated and characterized novel mouse mAb specific for a viral immunoevasin, the mouse CMV m12 protein, and suggest that these mAb may protect mice from CMV infection via passive immunity.

For the past two decades, the use of genetically fused fluorescent proteins (FPs) has greatly contributed to the study of chemotactic signalling in Escherichia coli including the activation of the response regulator protein CheY and its interaction with the flagellar motor. However, this approach suffers from a number of limitations, both biological and biophysical: for example, not all fusions are fully functional when fused to a bulky FP, which can have a similar molecular weight to its fused counterpart; they may interfere with the native interactions of the protein and the chromophores of FPs have low brightness and photostability and fast photobleaching rates. A recently developed technique for the electroporation of fluorescently labelled proteins in live bacteria has enabled us to bypass these limitations and study the in vivo behaviour of CheY at the single-molecule level. Here we show that purified CheY proteins labelled with organic dyes can be internalized into E. coli cells in controllable concentrations and imaged with video fluorescence microscopy. The use of this approach is illustrated by showing single CheY molecules diffusing within cells and interacting with the sensory clusters and the flagellar motors in real time. This article is part of the themed issue ‘The new bacteriology’.

This article considers the 2014 podcast Serial within a context of technical change and podcast histories. Building on previous work, the author argues that Serial is a significant moment in the history of podcasting. Recent trends have shifted podcasts from iPods to networked screen-based mobile devices in smartphones and connected dashboards, locations where global brands enjoy a significant advantage. It is in this context that the author places Serial as cultural object.

An important goal in synthetic biology is the assembly of biomimetic cell-like structures, which combine multiple biological components in synthetic lipid vesicles. A key limiting assembly step is the incorporation of membrane proteins into the lipid bilayer of the vesicles. Here we present a simple method for delivery of membrane proteins into a lipid bilayer within 5 min. Fusogenic proteoliposomes, containing charged lipids and membrane proteins, fuse with oppositely charged bilayers, with no requirement for detergent or fusion-promoting proteins, and deliver large, fragile membrane protein complexes into the target bilayers. We demonstrate the feasibility of our method by assembling a minimal electron transport chain capable of adenosine triphosphate (ATP) synthesis, combining Escherichia coli F1Fo ATP-synthase and the primary proton pump bo3-oxidase, into synthetic lipid vesicles with sizes ranging from 100 nm to ∼10 μm. This provides a platform for the combination of multiple sets of membrane protein complexes into cell-like artificial structures., Assembling multiple biological components into synthetic lipid vesicles is a limiting step in the manufacture of biomimetic cell-like structures. Here the authors use fusogenic proteoliposomes of opposite charge for fast assembly of a minimal electron transport chain consisting of F1F0 ATP-synthase and the proton pump bo3-oxidase.

Some proteins in biological complexes exchange with pools of free proteins while the complex is functioning. Evidence is emerging that protein exchange can be part of an adaptive mechanism. The bacterial flagellar motor is one of the most complex biological machines and is an ideal model system to study protein dynamics in large multimeric complexes. Recent studies showed that the copy number of FliM in the switch complex and the fraction of FliM that exchanges vary with the direction of flagellar rotation. Here, we investigated the stoichiometry and turnover of another switch complex component, FliN, labeled with the fluorescent protein CyPet, in Escherichia coli. Our results confirm that, in vivo, FliM and FliN form a complex with stoichiometry of 1:4 and function as a unit. We estimated that wild-type motors contained 120 ± 26 FliN molecules. Motors that rotated only clockwise (CW) or counterclockwise (CCW) contained 114 ± 17 and 144 ± 26 FliN molecules, respectively. The ratio of CCW-to-CW FliN copy numbers was 1.26, very close to that of 1.29 reported previously for FliM. We also measured the exchange of FliN molecules, which had a time scale and dependence upon rotation direction similar to those of FliM, consistent with an exchange of FliM-FliN as a unit. Our work confirms the highly dynamic nature of multimeric protein complexes and indicates that, under physiological conditions, these machines might not be the stable, complete structures suggested by averaged fixed methodologies but, rather, incomplete rings that can respond and adapt to changing environments., IMPORTANCE The flagellum is one of the most complex structures in a bacterial cell, with the core motor proteins conserved across species. Evidence is now emerging that turnover of some of these motor proteins depends on motor activity, suggesting that turnover is important for function. The switch complex transmits the chemosensory signal to the rotor, and we show, by using single-cell measurement, that both the copy number and the fraction of exchanging molecules vary with the rotational bias of the rotor. When the motor is locked in counterclockwise rotation, the copy number is similar to that determined by averaged, fixed methodologies, but when locked in a clockwise direction, the number is much lower, suggesting that that the switch complex ring is incomplete. Our results suggest that motor remodeling is an important component in tuning responses and adaptation at the motor.

A vegetation analysis reveals that the Lai Chi Wo mangrove swamp in Hong Kong has 10 mangrove species comprising 76.9% of the total number of mangrove species in Hong Kong. This swamp is a mixed community dominated byAegiceras corniculatum, Heritiera littoralis, Excoecaria agallocha, Kandelia candel, Bruguiera gymnorrhiza, with importance values of 37.88, 28.19, 14.33, 11.33 and 8.27 respectively. The community’s mangrove species diversity and evenness, as based on the Shannon-Wiener Function, were 1.44 and 62.01%, which were lower than those of the south su-tropical evergreen broad-leaf forest community but its ecological dominance (0.26) was higher. The correlation information field (CIF) method, a useful analytical method for complex forest community, was adopted in the present mangrove analysis. The species correlation information was:Aegiceras corniculatum (0.268)>Heritiera littoralis (0.250)>Kandelia candel (0.220)>Excoecaria agallocha (0.169)>Bruguiera gymnorrhiza (0.101). The correlation information of the mangrove community was 1.008.

In recent years, there has been a growing area of research focused on points of convergence between scientific and humanities discourses, with methods of interpreting cultural products drawing increasingly on other disciplines and vice versa. Ecological readings of cultural materials would be included here, ecological not in the traditional sense of environmental criticism, but rather focused on the elements of network relations as they play out within the living systems of cultural works. This chapter will examine the ways in which podcasts can share structural and epistemological affinities with ecological processes, engaging the conversational science podcast Stuff to Blow Your Mind (STBYM) as a case study. I will argue that STBYM, known for its elegantly produced discourse around complex material, with episodes like ‘Meet Your Bacterial Masters’ and ‘The Habitable Epoch’, exemplifies a growing trend toward epistemologically complex methods of approach to cultural processes.

Richard Herring is a writer, stand-up comedian and podcaster. In recent years he has found sustained success as a podcaster, particularly through the podcast Richard Herring’s Leicester Square Theatre Podcast (RHLSTP), on which he interviews a renowned comedian, broadcaster or celebrity guest in front of a live studio audience. RHLSTP is released in both audio and video versions. He also uses the podcast form to explore improvisational aspects of his writing and stand-up through the As It Occurs To Me (AIOTM) and Richard Herring’s Meaning Of Life (RHMOL) projects. Then there’s the surreal performance art of Me1 vs Me2 Snooker that finds Herring playing himself at snooker and taking on the persona of both players and also commentating on the frame as it happens, in audio form. Due to the variety of podcast forms he engages with and his high profile through them, Herring is an appropriate practitioner to provide a case study interview for this book. His work cuts across several types of podcast – live, pre-recorded, comedy, interview/chat etc. and podcasting is central to his work. It is the area where he gains the most recognition of the varied work he does and also, as he discusses in the interview, podcasting provides the opportunity to drive audiences and consumers to the other areas such as his stand-up shows. He is also a reflective practitioner taking the opportunity to discuss podcasting as a form on his RHLSTP podcast when a suitable opportunity arises