The BC Radon Data Repository (BCRDR) and BC Radon Map: Integrating disparate data sources for improved public health communication.

SETTING: The potential for exposure to indoor radon varies dramatically across British Columbia (BC) due to varied geology. Individuals may struggle to understand their exposure risk and agencies may struggle to understand the value of population-level programs and policies to mitigate risk. INTERVENTION: The BC Centre for Disease Control (BCCDC) established the BC Radon Data Repository (BCRDR) to facilitate radon research, public awareness, and action in the province. The BCRDR aggregates indoor radon measurements collected by government agencies, industry professionals and organizations, and research and advocacy groups. Participation was formalized with a data sharing agreement, which outlines how the BCCDC anonymizes and manages the shared data integrated into the BCRDR. OUTCOMES: The BCRDR currently holds 38,733 measurements from 18 data contributors. The repository continues to grow with new measurements from existing contributors and the addition of new contributors. A prominent use of the BCRDR was to create the online, interactive BC Radon Map, which includes regional concentration summaries, risk interpretation messaging, and health promotion information. Anonymized BCRDR data are also available for external release upon request. IMPLICATIONS: The BCCDC leverages existing radon measurement programs to create a large and integrated database with wide geographic coverage. The development and application of the BCRDR informs public health research and action beyond the BCCDC, and the repository can serve as a model for other regional or national initiatives.

RéSUMé: LIEU: Le potentiel d'exposition au radon à l'intérieur des bâtiments varie beaucoup d'une région à l'autre de la Colombie-Britannique en raison de la géologie variée. Les particuliers peuvent avoir du mal à comprendre leur risque d'exposition, et les organismes, à comprendre l'utilité des programmes et des politiques populationnels pour atténuer le risque. INTERVENTION: Le BC Centre for Disease Control (« le Centre ¼) a créé un organe d'archivage, le BC Radon Data Repository (BCRDR), pour faciliter la recherche, l'information, la sensibilisation du public et l'action liées au radon dans la province. Le BCRDR totalise les relevés du radon à l'intérieur des bâtiments pris par les organismes gouvernementaux, les professionnels et les organismes de l'industrie, ainsi que les groupes de recherche et de revendication. La participation est officialisée par un accord de partage de données qui décrit comment le Centre anonymise et gère les données communes du BCRDR. RéSULTATS: Le BCRDR contient actuellement 38 733 relevés de 18 contributeurs de données. Il continue de croître, avec de nouveaux relevés venant de contributeurs existants et l'ajout de nouveaux contributeurs. Il a servi, entre autres, à créer une carte du radon interactive en ligne pour la Colombie-Britannique, avec des résumés des concentrations régionales, des messages d'interprétation du risque et des informations de promotion de la santé. Sur demande, les données anonymisées du BCRDR sont également disponibles pour diffusion externe. CONSéQUENCES: Le Centre a exploité les programmes de prise de relevés du radon existants pour créer une grande base de données intégrée ayant une vaste couverture géographique. Le développement et les applications du BCRDR éclairent la recherche et l'action en santé publique au-delà du Centre, et l'organe d'archivage peut servir de modèle pour d'autres initiatives régionales ou nationales.

Public Health Messaging for Wildfire Smoke: Cast a Wide Net.

Wildfire smoke events are increasing in British Columbia (BC), Canada and environmental and public health agencies are responsible for communicating the health-related risks and mitigation strategies. To evaluate and identify opportunities for improving public communications about wildfire smoke and associated health risks we collaborated with end-users and developed a 32-question online survey. The survey was deployed province-wide from 29 September to 31 December 2020 following a severe wildfire smoke episode, which impacted large parts of BC. Using a convenience sample, we disseminated the survey through email lists, radio advertisements, a provincial research platform, and snowball methods. There were 757 respondents, who were generally representative of provincial demographics. Respondents indicated that they receive wildfire smoke messages from diverse sources, including: websites, social media, radio, and television. Radio was identified as the most important source of information for populations that may have increased exposure or health risks, including Indigenous respondents and those working in the trades. Respondents with lower educational attainment expressed that messaging should be simplified. Environmental and public health agencies should continue to share wildfire smoke messages using diverse methods, ideally tailoring the messages and methods to specific populations at risk for exposure and health effects.

Using Low-Cost Sensors to Assess Fine Particulate Matter Infiltration (PM2.5) during a Wildfire Smoke Episode at a Large Inpatient Healthcare Facility.

Wildfire smoke exposure is associated with a range of acute health outcomes, which can be more severe in individuals with underlying health conditions. Currently, there is limited information on the susceptibility of healthcare facilities to smoke infiltration. As part of a larger study to address this gap, a rehabilitation facility in Vancouver, Canada was outfitted with one outdoor and seven indoor low-cost fine particulate matter (PM2.5) sensors in Air Quality Eggs (EGG) during the summer of 2020. Raw measurements were calibrated using temperature, relative humidity, and dew point derived from the EGG data. The infiltration coefficient was quantified using a distributed lag model. Indoor concentrations during the smoke episode were elevated throughout the building, though non-uniformly. After censoring indoor-only peaks, the average infiltration coefficient (range) during typical days was 0.32 (0.22-0.39), compared with 0.37 (0.31-0.47) during the smoke episode, a 19% increase on average. Indoor PM2.5 concentrations quickly reflected outdoor conditions during and after the smoke episode. It is unclear whether these results will be generalizable to other years due to COVID-related changes to building operations, but some of the safety protocols may offer valuable lessons for future wildfire seasons. For example, points of building entry and exit were reduced from eight to two during the pandemic, which likely helped to protect the building from wildfire smoke infiltration. Overall, these results demonstrate the utility of indoor low-cost sensors in understanding the impacts of extreme smoke events on facilities where highly susceptible individuals are present. Furthermore, they highlight the need to employ interventions that enhance indoor air quality in such facilities during smoke events.

Binding interface and impact on protease cleavage for an RNA aptamer to HIV-1 reverse transcriptase.

RNA aptamers that bind HIV-1 reverse transcriptase (RT) inhibit RT in enzymatic and viral replication assays. Some aptamers inhibit RT from only a few viral clades, while others show broad-spectrum inhibition. Biophysical determinants of recognition specificity are poorly understood. We investigated the interface between HIV-1 RT and a broad-spectrum UCAA-family aptamer. SAR and hydroxyl radical probing identified aptamer structural elements critical for inhibition and established the role of signature UCAA bulge motif in RT-aptamer interaction. HDX footprinting on RT ± aptamer shows strong contacts with both subunits, especially near the C-terminus of p51. Alanine scanning revealed decreased inhibition by the aptamer for mutants P420A, L422A and K424A. 2D proton nuclear magnetic resonance and SAXS data provided constraints on the solution structure of the aptamer and enable computational modeling of the docked complex with RT. Surprisingly, the aptamer enhanced proteolytic cleavage of precursor p66/p66 by HIV-1 protease, suggesting that it stabilizes the productive conformation to allow maturation. These results illuminate features at the RT-aptamer interface that govern recognition specificity by a broad-spectrum antiviral aptamer, and they open new possibilities for accelerating RT maturation and interfering with viral replication.

Poly-Target Selection Identifies Broad-Spectrum RNA Aptamers.

Aptamer selections often yield distinct subpopulations, each with unique phenotypes that can be leveraged for specialized applications. Although most selections aim to attain ever higher specificity, we sought to identify aptamers that recognize increasingly divergent primate lentiviral reverse transcriptases (RTs). We hypothesized that aptamer subpopulations in libraries pre-enriched against a single RT may exhibit broad-spectrum binding and inhibition, and we devised a multiplexed poly-target selection to elicit those phenotypes against a panel of primate lentiviral RTs. High-throughput sequencing and coenrichment/codepletion analysis of parallel and duplicate selection trajectories rapidly narrowed the list of candidate aptamers by orders of magnitude and identified dozens of priority candidates for further screening. Biochemical characterization validated a novel aptamer motif and several rare and unobserved variants of previously known motifs that inhibited recombinant RTs to varying degrees. These broad-spectrum aptamers also suppressed replication of viral constructs carrying phylogenetically diverse RTs. The poly-target selection and coenrichment/codepletion approach described herein is a generalizable strategy for identifying cross-reactivity among related targets from combinatorial libraries.

RNA-protein interactions govern antiviral specificity and encapsidation of broad spectrum anti-HIV reverse transcriptase aptamers.

RNA aptamers that bind HIV-1 reverse transcriptase (RT) inhibit HIV-1 replication, but little is known about potential aptamer-specific viral resistance. During replication, RT interacts with diverse nucleic acids. Thus, the genetic threshold for eliciting resistance may be high for aptamers that make numerous contacts with RT. To evaluate the impact of RT-aptamer binding specificity on replication, we engineered proviral plasmids encoding diverse RTs within the backbone of HIV-1 strain NL4-3. Viruses inhibited by pseudoknot aptamers were rendered insensitive by a naturally occurring R277K variant, providing the first demonstration of aptamer-specific resistance in cell culture. Naturally occurring, pseudoknot-insensitive viruses were rendered sensitive by the inverse K277R mutation, establishing RT as the genetic locus for aptamer-mediated HIV-1 inhibition. Non-pseudoknot RNA aptamers exhibited broad-spectrum inhibition. Inhibition was observed only when virus was produced in aptamer-expressing cells, indicating that encapsidation is required. HIV-1 suppression magnitude correlated with the number of encapsidated aptamer transcripts per virion, with saturation occurring around 1:1 stoichiometry with packaged RT. Encapsidation specificity suggests that aptamers may encounter dimerized GagPol in the cytosol during viral assembly. This study provides new insights into HIV-1's capacity to escape aptamer-mediated inhibition, the potential utility of broad-spectrum aptamers to overcome resistance, and molecular interactions that occur during viral assembly.

Nucleobase modification by an RNA enzyme.

Ribozymes can catalyze phosphoryl or nucleotidyl transfer onto ribose hydroxyls of RNA chains. We report a single ribozyme that performs both reactions, with a nucleobase serving as initial acceptor moiety. This unprecedented combined reaction was revealed while investigating potential contributions of ribose hydroxyls to catalysis by kinase ribozyme K28. For a 58nt, cis-acting form of K28, each nucleotide could be replaced with the corresponding 2΄F analog without loss of activity, indicating that no particular 2΄OH is specifically required. Reactivities of two-stranded K28 variants with oligodeoxynucleotide acceptor strands devoid of any 2΄OH moieties implicate modification on an internal guanosine N-2, rather than a ribose hydroxyl. Product mass suggests formation of a GDP(S) adduct along with a second thiophosphorylation, implying that the ribozyme catalyzes both phosphoryl and nucleotidyl transfers. This is further supported by transfer of radiolabels into product from both α and γ phosphates of donor molecules. Furthermore, periodate reactivity of the final product signifies acquisition of a ribose sugar with an intact 2΄-3΄ vicinal diol. Neither nucleobase modification nor nucleotidyl transfer has previously been reported for a kinase ribozyme, making this a first-in-class ribozyme. Base-modifying ribozymes may have played important roles in early RNA world evolution by enhancing nucleic acid functions.