Intracellular proteomics and extracellular vesiculomics as a metric of disease recapitulation in 3D-bioprinted aortic valve arrays

In calcific aortic valve disease (CAVD), mechanosensitive valvular cells respond to fibrosis- and calcification-induced tissue stiffening, further driving pathophysiology. No pharmacotherapeutics are available to treat CAVD because of the paucity of (i) appropriate experimental models that recapitulate this complex environment and (ii) benchmarking novel engineered aortic valve (AV)–model performance. We established a biomaterial-based CAVD model mimicking the biomechanics of the human AV disease-prone fibrosa layer, three-dimensional (3D)–bioprinted into 96-well arrays. Liquid chromatography–tandem mass spectrometry analyses probed the cellular proteome and vesiculome to compare the 3D-bioprinted model versus traditional 2D monoculture, against human CAVD tissue. The 3D-bioprinted model highly recapitulated the CAVD cellular proteome (94% versus 70% of 2D proteins). Integration of cellular and vesicular datasets identified known and unknown proteins ubiquitous to AV calcification. This study explores how 2D versus 3D-bioengineered systems recapitulate unique aspects of human disease, positions multiomics as a technique for the evaluation of high throughput–based bioengineered model systems, and potentiates future drug discovery

Changes in symptomatology, reinfection, and transmissibility associated with the SARS-CoV-2 variant B.1.1.7: an ecological study

Background The SARS-CoV-2 variant B.1.1.7 was first identified in December, 2020, in England. We aimed to investigate whether increases in the proportion of infections with this variant are associated with differences in symptoms or disease course, reinfection rates, or transmissibility. Methods We did an ecological study to examine the association between the regional proportion of infections with the SARS-CoV-2 B.1.1.7 variant and reported symptoms, disease course, rates of reinfection, and transmissibility. Data on types and duration of symptoms were obtained from longitudinal reports from users of the COVID Symptom Study app who reported a positive test for COVID-19 between Sept 28 and Dec 27, 2020 (during which the prevalence of B.1.1.7 increased most notably in parts of the UK). From this dataset, we also estimated the frequency of possible reinfection, defined as the presence of two reported positive tests separated by more than 90 days with a period of reporting no symptoms for more than 7 days before the second positive test. The proportion of SARS-CoV-2 infections with the B.1.1.7 variant across the UK was estimated with use of genomic data from the COVID-19 Genomics UK Consortium and data from Public Health England on spike-gene target failure (a non-specific indicator of the B.1.1.7 variant) in community cases in England. We used linear regression to examine the association between reported symptoms and proportion of B.1.1.7. We assessed the Spearman correlation between the proportion of B.1.1.7 cases and number of reinfections over time, and between the number of positive tests and reinfections. We estimated incidence for B.1.1.7 and previous variants, and compared the effective reproduction number, Rt, for the two incidence estimates. Findings From Sept 28 to Dec 27, 2020, positive COVID-19 tests were reported by 36 920 COVID Symptom Study app users whose region was known and who reported as healthy on app sign-up. We found no changes in reported symptoms or disease duration associated with B.1.1.7. For the same period, possible reinfections were identified in 249 (0·7% [95% CI 0·6–0·8]) of 36 509 app users who reported a positive swab test before Oct 1, 2020, but there was no evidence that the frequency of reinfections was higher for the B.1.1.7 variant than for pre-existing variants. Reinfection occurrences were more positively correlated with the overall regional rise in cases (Spearman correlation 0·56–0·69 for South East, London, and East of England) than with the regional increase in the proportion of infections with the B.1.1.7 variant (Spearman correlation 0·38–0·56 in the same regions), suggesting B.1.1.7 does not substantially alter the risk of reinfection. We found a multiplicative increase in the Rt of B.1.1.7 by a factor of 1·35 (95% CI 1·02–1·69) relative to pre-existing variants. However, Rt fell below 1 during regional and national lockdowns, even in regions with high proportions of infections with the B.1.1.7 variant. Interpretation The lack of change in symptoms identified in this study indicates that existing testing and surveillance infrastructure do not need to change specifically for the B.1.1.7 variant. In addition, given that there was no apparent increase in the reinfection rate, vaccines are likely to remain effective against the B.1.1.7 variant. Funding Zoe Global, Department of Health (UK), Wellcome Trust, Engineering and Physical Sciences Research Council (UK), National Institute for Health Research (UK), Medical Research Council (UK), Alzheimer's Society

Genomic assessment of quarantine measures to prevent SARS-CoV-2 importation and transmission

Mitigation of SARS-CoV-2 transmission from international travel is a priority. We evaluated the effectiveness of travellers being required to quarantine for 14-days on return to England in Summer 2020. We identified 4,207 travel-related SARS-CoV-2 cases and their contacts, and identified 827 associated SARS-CoV-2 genomes. Overall, quarantine was associated with a lower rate of contacts, and the impact of quarantine was greatest in the 16–20 age-group. 186 SARS-CoV-2 genomes were sufficiently unique to identify travel-related clusters. Fewer genomically-linked cases were observed for index cases who returned from countries with quarantine requirement compared to countries with no quarantine requirement. This difference was explained by fewer importation events per identified genome for these cases, as opposed to fewer onward contacts per case. Overall, our study demonstrates that a 14-day quarantine period reduces, but does not completely eliminate, the onward transmission of imported cases, mainly by dissuading travel to countries with a quarantine requirement

Genomic epidemiology of SARS-CoV-2 in a UK university identifies dynamics of transmission

AbstractUnderstanding SARS-CoV-2 transmission in higher education settings is important to limit spread between students, and into at-risk populations. In this study, we sequenced 482 SARS-CoV-2 isolates from the University of Cambridge from 5 October to 6 December 2020. We perform a detailed phylogenetic comparison with 972 isolates from the surrounding community, complemented with epidemiological and contact tracing data, to determine transmission dynamics. We observe limited viral introductions into the university; the majority of student cases were linked to a single genetic cluster, likely following social gatherings at a venue outside the university. We identify considerable onward transmission associated with student accommodation and courses; this was effectively contained using local infection control measures and following a national lockdown. Transmission clusters were largely segregated within the university or the community. Our study highlights key determinants of SARS-CoV-2 transmission and effective interventions in a higher education setting that will inform public health policy during pandemics.</jats:p

Owyhee River intracanyon lava flows: Does the river give a dam?

Rivers carved into uplifted plateaus are commonly disrupted by discrete events from the surrounding landscape, such as lava flows or large mass movements. These disruptions are independent of slope, basin area, or channel discharge, and can dominate aspects of valley morphology and channel behavior for many kilometers. We document and assess the effects of one type of disruptive event, lava dams, on river valley morphology and incision rates at a variety of time scales, using examples from the Owyhee River in southeastern Oregon. Six sets of basaltic lava flows entered and dammed the river canyon during two periods in the late Cenozoic ca. 2 Ma–780 ka and 250–70 ka. The dams are strongly asymmetric, with steep, blunt escarpments facing up valley and long, low slopes down valley. None of the dams shows evidence of catastrophic failure; all blocked the river and diverted water over or around the dam crest. The net effect of the dams was therefore to inhibit rather than promote incision. Once incision resumed, most of the intracanyon flows were incised relatively rapidly and therefore did not exert a lasting impact on the river valley profile over time scales \u3e106 yr. The net long-term incision rate from the time of the oldest documented lava dam, the Bogus Rim lava dam (≤1.7 Ma), to present was 0.18 mm/yr, but incision rates through or around individual lava dams were up to an order of magnitude greater. At least three lava dams (Bogus Rim, Saddle Butte, and West Crater) show evidence that incision initiated only after the impounded lakes filled completely with sediment and there was gravel transport across the dams. The most recent lava dam, formed by the West Crater lava flow around 70 ka, persisted for at least 25 k.y. before incision began, and the dam was largely removed within another 35 k.y. The time scale over which the lava dams inhibit incision is therefore directly affected by both the volume of lava forming the dam and the time required for sediment to fill the blocked valley. Variations in this primary process of incision through the lava dams could be influenced by additional independent factors such as regional uplift, drainage integration, or climate that affect the relative base level, discharge, and sediment yield within the watershed. By redirecting the river, tributaries, and subsequent lava flows to different parts of the canyon, lava dams create a distinct valley morphology of flat, broad basalt shelves capping steep cliffs of Tertiary sediment. This stratigraphy is conducive to landsliding and extends the effects of intracanyon lava flows on channel geomorphology beyond the lifetime of the dams

"Just not all ice users do that": Investigating perceptions and potential harms of Australia's Ice Destroys Lives campaign in two studies

Background: In 2015, the Australian government launched the media campaign Ice Destroys Lives targeting crystal methamphetamine use. Previous research indicates mass media campaigns may have harmful effects for people engaged in drug use. This study investigated perceptions and harms of Ice Destroys Lives among adults with a history of injecting drugs and young people. Methods: This analysis includes data from two studies: an online questionnaire with young people and in-depth interviews with adults who use crystal methamphetamine. Young people from Victoria, Australia, were recruited through Facebook. We collected data on drug use, campaign recognition and behaviours. Participants who recognised the campaign indicated whether they agreed with five statements related to Ice Destroys Lives. We compared campaign perceptions between young people who reported ever using crystal methamphetamine and those who did not. Adults who use crystal methamphetamine were sampled from the Melbourne injecting drug user cohort study. We asked participants if they recognised the campaign and whether it represented their experiences. Results: One thousand twenty-nine young people completed the questionnaire; 71% were female, 4% had used crystal methamphetamine and 69% recognised Ice Destroys Lives. Three quarters agreed the campaign made them not want to use ice. Ever using crystal methamphetamine was associated with disagreeing with three statements including this campaign makes you not want to use ice (adjusted odds ratio (AOR) = 4.3, confidence interval (CI) = 1.8-10.0), this campaign accurately portrays the risks of ice use (AOR = 3.2, CI = 1.4-7.6) and this campaign makes you think that people who use ice are dangerous (AOR = 6.6, CI = 2.2-19.8). We interviewed 14 people who used crystal methamphetamine; most were male, aged 29-39 years, and most recognised the campaign. Participants believed Ice Destroys Lives misrepresented their experiences and exaggerated "the nasty side" of drug use. Participants felt the campaign exacerbated negative labels and portrayed people who use crystal methamphetamine as "violent" and "crazy". Conclusion: In our study, Ice Destroys Lives was widely recognised and delivered a prevention message to young people. However, for people with a history of crystal methamphetamine use, the campaign also reinforced negative stereotypes and did not encourage help seeking. Alternative evidence-based strategies are required to reduce crystal methamphetamine-related harms

Genomic reconstruction of the SARS-CoV-2 epidemic in England

AbstractThe evolution of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus leads to new variants that warrant timely epidemiological characterization. Here we use the dense genomic surveillance data generated by the COVID-19 Genomics UK Consortium to reconstruct the dynamics of 71 different lineages in each of 315 English local authorities between September 2020 and June 2021. This analysis reveals a series of subepidemics that peaked in early autumn 2020, followed by a jump in transmissibility of the B.1.1.7/Alpha lineage. The Alpha variant grew when other lineages declined during the second national lockdown and regionally tiered restrictions between November and December 2020. A third more stringent national lockdown suppressed the Alpha variant and eliminated nearly all other lineages in early 2021. Yet a series of variants (most of which contained the spike E484K mutation) defied these trends and persisted at moderately increasing proportions. However, by accounting for sustained introductions, we found that the transmissibility of these variants is unlikely to have exceeded the transmissibility of the Alpha variant. Finally, B.1.617.2/Delta was repeatedly introduced in England and grew rapidly in early summer 2021, constituting approximately 98% of sampled SARS-CoV-2 genomes on 26 June 2021.</jats:p