The clinical utility of sequencing the entirety of CFTR.

BACKGROUND: Cystic fibrosis (CF) is caused by deleterious variants in each CFTR gene. We investigated the utility of whole-gene CFTR sequencing when fewer than two pathogenic or likely pathogenic (P/LP) variants were detected by conventional testing (sequencing of exons and flanking introns) of CFTR. METHODS: Individuals with features of CF and a CF-diagnostic sweat chloride concentration with zero or one P/LP variants identified by conventional testing enrolled in the CF Mutation Analysis Program (MAP) underwent whole-gene CFTR sequencing. Replication was performed on individuals enrolled in the CF Genome Project (CFGP), followed by phenotype review and interrogation of other genes. RESULTS: Whole-gene sequencing identified a second P/LP variant in 20/43 MAP enrollees (47 %) and 10/22 CFGP enrollees (45 %) who had one P/LP variant after conventional testing. No P/LP variants were detected when conventional testing was negative (MAP: n = 43; CFGP: n = 13). Genome-wide analysis was unable to find an alternative etiology in CFGP participants with fewer than two P/LP CFTR variants and CF could not be confirmed in 91 % following phenotype re-review. CONCLUSIONS: Whole-gene CFTR analysis is beneficial in individuals with one previously-identified P/LP variant and a CF-diagnostic sweat chloride. Negative conventional CFTR testing indicates that the phenotype should be re-evaluated.

Engineering nanowires in bacteria to elucidate electron transport structural-functional relationships.

Bacterial pilin nanowires are protein complexes, suggested to possess electroactive capabilities forming part of the cells' bioenergetic programming. Their role is thought to be linked to facilitating electron transfer between cells and the external environment to permit metabolism and cell-to-cell communication. There is a significant debate, with varying hypotheses as to the nature of the proteins currently lying between type-IV pilin-based nanowires and polymerised cytochrome-based filaments. Importantly, to date, there is a very limited structure-function analysis of these structures within whole bacteria. In this work, we engineered Cupriavidus necator H16, a model autotrophic organism to express differing aromatic modifications of type-IV pilus proteins to establish structure-function relationships on conductivity and the effects this has on pili structure. This was achieved via a combination of high-resolution PeakForce tunnelling atomic force microscopy (PeakForce TUNA™) technology, alongside conventional electrochemical approaches enabling the elucidation of conductive nanowires emanating from whole bacterial cells. This work is the first example of functional type-IV pili protein nanowires produced under aerobic conditions using a Cupriavidus necator chassis. This work has far-reaching consequences in understanding the basis of bio-electrical communication between cells and with their external environment.

Self-Reported Effects of the Covid-19 Pandemic on Stewardship Organizations and Their Activities in Southeast New England, USA.

In many communities, regions, or landscapes, there are numerous environmental groups working across different sectors and creating stewardship networks that shape the environment and the benefits people derive from it. The make-up of these networks can vary, but generally include organizations of different sizes and capacities. As the Covid-19 pandemic (2020 to the present) shuts down businesses and nonprofits, catalyzes new initiatives, and generally alters the day-to-day professional and personal lives, it is logical to assume that these stewardship networks and their environmental work are impacted; exactly how, is unknown. In this study, we analyze the self-reported effects of the Covid-19 pandemic on stewardship groups working in southeast New England, USA. Stewardship organizations were surveyed from November 2020 to April 2021 and asked, among other questions, "How is Covid-19 affecting your organization?" We analyzed responses using several qualitative coding approaches. Our analysis revealed group-level impacts including changes in group capacity, challenges in managing access to public green spaces, and altered forms of volunteer engagement. These results provide insights into the varied effects of the Covid-19 pandemic and government responses such as stay-at-home orders and social distancing policies on stewardship that can inform the development of programs to reduce negative outcomes and enhance emerging capacities and innovations.

Affliction And Resignation in George Herbert: Reflections on Human Agency In A Global Pandemic.

This paper reflects on the context of lockdown in a global pandemic, where entire populations have experienced severe curtailments of the opportunity to exercise agency. The experience has made me notice the surprisingly large role that resignation has played in Christian moral thinking in earlier ages. This paper takes the devotional poetry of George Herbert as a case study, since Herbert had a particular preoccupation with the psychology of religious belief in circumstances where the will cannot be deployed to any end. Herbert reflects on the predicament in which the moral life has to continue even though opportunities for the exercise of agency have collapsed. After examining Herbert's poems of affliction, I conclude by setting out a brief case for understanding resignation as a moral practice - a practice that centres not on the will but on imagination and the emotions.