COVID-19 during pregnancy alters circulating extracellular vesicle cargo and their effects on trophoblast (preprint)

SARS-CoV-2 infection and the resulting coronavirus disease (COVID-19) complicate pregnancies as the result of placental dysfunction which increases the risk of adverse pregnancy outcomes. While abnormal placental pathology resulting from COVID-19 is common, direct infection of the placenta is rare. This suggests maternal response to infection is responsible for placental dysfunction. We hypothesized that maternal circulating extracellular vesicles (EVs) are altered by COVID-19 during pregnancy and contribute to placental dysfunction. To examine this, we characterized maternal circulating EVs from pregnancies complicated by COVID-19 and tested their functional effect on trophoblast cells in vitro. We found the timing of infection is a major determinant of the effect of COVID-19 on circulating EVs. Additionally, we found differentially expressed EV mRNA cargo in COVID-19 groups compared to Controls that regulates the differential gene expression induced by COVID-19 in the placenta. In vitro exposure of trophoblasts to EVs isolated from patients with an active infection, but not EVs isolated from Controls, reduced key trophoblast functions including hormone production and invasion. This demonstrates circulating EVs from subjects with an active infection disrupt vital trophoblast function. This study determined that COVID-19 has a long-lasting effect on circulating EVs and circulating EVs are likely to participate in the placental dysfunction induced by COVID-19.

The Untold Stories from COVID-19: The Calamity of Implementing Curriculum during a Pandemic

In March 2020, the COVID-19 Pandemic wreaked havoc on our nation's educational system. Students, teachers, and administrators were forced to engage in a new remote learning model, which was unfamiliar. This narrative study draws on the lived experiences of six K-12 teachers in Southwest Ohio urban school districts. The data analysis was examined through the lens of the Science of Learning and Development framework (SoLD). Findings highlight the impact of COVID-19 on curriculum implementation. Results show that unprepared teachers could not pivot to online learning effectively, which may intensify the educational gaps and inequities among students in six urban schools in Southwest Ohio.

Sleep disturbances and depression are bidirectionally associated among college student athletes across COVID-19 pandemic exposure classes.

College athletes may be vulnerable to sleep disturbances and depression during the COVID-19 pandemic as a result of large shifts in social and athletic obligations. In a national sample of college athletes, we examined the associations between sleep disturbances and depression across two timepoints, using COVID-19 exposure as a moderator. Data were collected from 2098 NCAA Division I, II, and III college athletes during two timepoints, from April 10 to May 23, and from August 4 to September 15, 2020. First, a latent class analysis was conducted with five indicators of levels of COVID-19 exposure to determine different exposure profiles. Second, to examine the directionality of associations between sleep disturbance and depression, a cross-lagged panel model was added to the latent class membership structural equation model; this allowed for testing of moderation by COVID exposure class membership. Four highly homogeneous, well-separated classes of COVID-19 exposure were enumerated: Low Exposure (57%); Quarantine Only (21%); High Other, Low Self Exposure (14%); and High Exposure (8%). COVID-19 exposure class membership did not significantly moderate associations between sleep disturbances and depression. However, student athletes significantly differed in T2 depression by their COVID-19 exposure class membership. Depression and sleep disturbances were positively correlated at both timepoints (r T1 = 0.39; r T2 = 0.30). Additionally, cross-lagged associations were found such that T2 depression was associated with T1 sleep disturbances (ß = 0.14) and vice versa (ß = 0.11). These cross-lagged associations were not significantly affected by athletes' level of COVID-19 exposure during the beginning of the pandemic.

Thiopurines inhibit coronavirus Spike protein processing and incorporation into progeny virions.

There is an outstanding need for broadly acting antiviral drugs to combat emerging viral diseases. Here, we report that thiopurines inhibit the replication of the betacoronaviruses HCoV-OC43 and SARS-CoV-2. 6-Thioguanine (6-TG) disrupted early stages of infection, limiting accumulation of full-length viral genomes, subgenomic RNAs and structural proteins. In ectopic expression models, we observed that 6-TG increased the electrophoretic mobility of Spike from diverse betacoronaviruses, matching the effects of enzymatic removal of N-linked oligosaccharides from Spike in vitro. SARS-CoV-2 virus-like particles (VLPs) harvested from 6-TG-treated cells were deficient in Spike. 6-TG treatment had a similar effect on production of lentiviruses pseudotyped with SARS-CoV-2 Spike, yielding pseudoviruses deficient in Spike and unable to infect ACE2-expressing cells. Together, these findings from complementary ectopic expression and infection models strongly indicate that defective Spike trafficking and processing is an outcome of 6-TG treatment. Using biochemical and genetic approaches we demonstrated that 6-TG is a pro-drug that must be converted to the nucleotide form by hypoxanthine phosphoribosyltransferase 1 (HPRT1) to achieve antiviral activity. This nucleotide form has been shown to inhibit small GTPases Rac1, RhoA, and CDC42; however, we observed that selective chemical inhibitors of these GTPases had no effect on Spike processing or accumulation. By contrast, the broad GTPase agonist ML099 countered the effects of 6-TG, suggesting that the antiviral activity of 6-TG requires the targeting of an unknown GTPase. Overall, these findings suggest that small GTPases are promising targets for host-targeted antivirals.

RAMPVIS: Answering the challenges of building visualisation capabilities for large-scale emergency responses.

The effort for combating the COVID-19 pandemic around the world has resulted in a huge amount of data, e.g., from testing, contact tracing, modelling, treatment, vaccine trials, and more. In addition to numerous challenges in epidemiology, healthcare, biosciences, and social sciences, there has been an urgent need to develop and provide visualisation and visual analytics (VIS) capacities to support emergency responses under difficult operational conditions. In this paper, we report the experience of a group of VIS volunteers who have been working in a large research and development consortium and providing VIS support to various observational, analytical, model-developmental, and disseminative tasks. In particular, we describe our approaches to the challenges that we have encountered in requirements analysis, data acquisition, visual design, software design, system development, team organisation, and resource planning. By reflecting on our experience, we propose a set of recommendations as the first step towards a methodology for developing and providing rapid VIS capacities to support emergency responses.

Thiopurines inhibit coronavirus Spike protein processing and incorporation into progeny virions (preprint)

There is an outstanding need for broadly acting antiviral drugs to combat emerging viral diseases. Here, we report that thiopurines inhibit the replication of the betacoronaviruses HCoV-OC43 and SARS-CoV-2, and to a lesser extent, the alphacoronavirus HCoV-229E. 6-Thioguanine (6-TG) disrupted early stages of infection, limiting synthesis of full-length and subgenomic HCoV RNAs. Furthermore, consistent with our previous report on the effects of thiopurines on influenza A virus (IAV) glycoproteins, we observed that 6-TG inhibited accumulation of Spike glycoproteins from diverse HCoVs. Specifically, 6-TG treatment decreased the accumulation of Spike proteins and increased their electrophoretic mobility to match the properties of Spike following enzymatic removal of N-linked oligosaccharides with Peptide:N-glycosidase F (PNGaseF). SARS-CoV-2 virus-like particles (VLPs) harvested from 6-TG-treated cells were deficient in Spike. 6-TG treatment had a similar effect on lentiviruses pseudotyped with SARS-CoV-2 Spike; lentiviruses could be harvested from cell supernatants, but they were deficient in Spike and unable to infect human cells bearing ACE2 receptors. Together, these findings from complementary ectopic expression and infection models strongly indicate that defective Spike trafficking and processing is an outcome of 6-TG treatment. At low micromolar doses, the primary known mode of action of 6-TG is selective inhibition of the small GTPase Rac1. However, we show that selective chemical inhibitors of the small GTPases Rac1, CDC42 and Rho had no effect on Spike processing and accumulation, whereas the broad GTPase agonist ML099 was able to counter the effects of 6-TG, suggesting that an unknown GTPase could be the relevant 6-TG-target protein involved in regulating Spike processing and accumulation. Overall, these findings provide important clues about the mechanism of action of a candidate antiviral that can broadly target HCoVs and suggest that small GTPases may be promising targets for host-targeted antivirals.

Appropriate Telemedicine Utilization in Spine Surgery: Results From a Delphi Study.

STUDY DESIGN: Delphi expert panel consensus. OBJECTIVE: To obtain expert consensus on best practices for appropriate telemedicine utilization in spine surgery. SUMMARY OF BACKGROUND DATA: Several studies have shown high patient satisfaction associated with telemedicine during the COVID-19 peak pandemic period as well as after easing of restrictions. As this technology will most likely continue to be employed, there is a need to define appropriate utilization. METHODS: An expert panel consisting of 27 spine surgeons from various countries was assembled in February 2021. A two-round consensus-based Delphi method was used to generate consensus statements on various aspects of telemedicine (separated as video visits or audio visits) including themes, such as patient location and impact of patient diagnosis, on assessment of new patients. Topics with ≥75% agreement were categorized as having achieved a consensus. RESULTS: The expert panel reviewed a total of 59 statements. Of these, 32 achieved consensus. The panel had consensus that video visits could be utilized regardless of patient location and that video visits are appropriate for evaluating as well as indicating for surgery multiple common spine pathologies, such as lumbar stenosis, lumbar radiculopathy, and cervical radiculopathy. Finally, the panel had consensus that video visits could be appropriate for a variety of visit types including early, midterm, longer term postoperative follow-up, follow-up for imaging review, and follow-up after an intervention (i.e., physical therapy, injection). CONCLUSION: Although telemedicine was initially introduced out of necessity, this technology most likely will remain due to evidence of high patient satisfaction and significant cost savings. This study was able to provide a framework for appropriate telemedicine utilization in spine surgery from a panel of experts. However, several questions remain for future research, such as whether or not an in-person consultation is necessary prior to surgery and which physical exam maneuvers are appropriate for telemedicine.Level of Evidence: 4.

COVID-19 Lockdowns: Exacerbating the Silent Pandemic

The global medical community has exalted the vaccine as the champion solution to end the violent toll inflicted by COVID-19. While the role of vaccines cannot be undervalued in wide-scale intervention, presenting them as the sole solution exonerates individuals of the importance of taking ownership over their lifestyle choices. This editorial focuses on the importance of physical activity as a crucial component of COVID-19 prevention programs and a long-term investment against chronic diseases.

One Health Approach: A Data-Driven Priority for Mitigating Outbreaks of Emerging and Re-Emerging Zoonotic Infectious Diseases.

This paper discusses the contributions that One Health principles can make in improving global response to zoonotic infectious disease. We highlight some key benefits of taking a One Health approach to a range of complex infectious disease problems that have defied a more traditional sectoral approach, as well as public health policy and practice, where gaps in surveillance systems need to be addressed. The historical examples demonstrate the scope of One Health, partly from an Australian perspective, but also with an international flavour, and illustrate innovative approaches and outcomes with the types of collaborative partnerships that are required.

Photodynamic Inactivation of Human Coronaviruses.

Photodynamic inactivation (PDI) employs a photosensitizer, light, and oxygen to create a local burst of reactive oxygen species (ROS) that can inactivate microorganisms. The botanical extract PhytoQuinTM is a powerful photosensitizer with antimicrobial properties. We previously demonstrated that photoactivated PhytoQuin also has antiviral properties against herpes simplex viruses and adenoviruses in a dose-dependent manner across a broad range of sub-cytotoxic concentrations. Here, we report that human coronaviruses (HCoVs) are also susceptible to photodynamic inactivation. Photoactivated-PhytoQuin inhibited the replication of the alphacoronavirus HCoV-229E and the betacoronavirus HCoV-OC43 in cultured cells across a range of sub-cytotoxic doses. This antiviral effect was light-dependent, as we observed minimal antiviral effect of PhytoQuin in the absence of photoactivation. Using RNase protection assays, we observed that PDI disrupted HCoV particle integrity allowing for the digestion of viral RNA by exogenous ribonucleases. Using lentiviruses pseudotyped with the SARS-CoV-2 Spike (S) protein, we once again observed a strong, light-dependent antiviral effect of PhytoQuin, which prevented S-mediated entry into human cells. We also observed that PhytoQuin PDI altered S protein electrophoretic mobility. The PhytoQuin constituent emodin displayed equivalent light-dependent antiviral activity to PhytoQuin in matched-dose experiments, indicating that it plays a central role in PhytoQuin PDI against CoVs. Together, these findings demonstrate that HCoV lipid envelopes and proteins are damaged by PhytoQuin PDI and expands the list of susceptible viruses.

A serum-stable RNA aptamer specific for SARS-CoV-2 neutralizes viral entry.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has created an urgent need for new technologies to treat COVID-19. Here we report a 2'-fluoro protected RNA aptamer that binds with high affinity to the receptor binding domain (RBD) of SARS-CoV-2 spike protein, thereby preventing its interaction with the host receptor ACE2. A trimerized version of the RNA aptamer matching the three RBDs in each spike complex enhances binding affinity down to the low picomolar range. Binding mode and specificity for the aptamer-spike interaction is supported by biolayer interferometry, single-molecule fluorescence microscopy, and flow-induced dispersion analysis in vitro. Cell culture experiments using virus-like particles and live SARS-CoV-2 show that the aptamer and, to a larger extent, the trimeric aptamer can efficiently block viral infection at low concentration. Finally, the aptamer maintains its high binding affinity to spike from other circulating SARS-CoV-2 strains, suggesting that it could find widespread use for the detection and treatment of SARS-CoV-2 and emerging variants.

Relationships between Physical Activity, Work Ability, Absenteeism and Presenteeism in Australian and New Zealand Adults during COVID-19.

Public health movement and social restrictions imposed by the Australian and New Zealand governments in response to the COVID-19 pandemic influenced the working environment and may have affected health behaviours, work ability, and job performance. The aim of this study was to determine the associations between health behaviours and work ability and performance during COVID-19 restrictions and if health behaviours were related to demographic or population factors. A cross-sectional survey was used to gather responses from 433 adult employees in Australia and New Zealand between June and August 2020. The survey requested demographic information and used the International Physical Activity Questionnaire, Work Ability Index, and the World Health Organisation's Health and Work Performance Questionnaire. Multivariate regression models were used to explore relationships between the identified variables while controlling for several possible confounders. Being sufficiently physically active was associated with higher reported physical (aOR = 2.1; p = 0.001) and mental work abilities (aOR = 1.8; p = 0.007) and self-reported job performance (i.e., lower presenteeism) (median +7.42%; p = 0.03). Part-time employees were 56% less likely (p = 0.002) to report a good or very good mental work ability. Those with existing medical conditions were 14% less likely (p = 0.008) to be sufficiently active and 80% less likely (p = 0.002) to report rather good or very good physical work ability. Being sufficiently active was associated with higher physical and mental work abilities and better job performance during the COVID-19 pandemic. Employers should support opportunities for regular physical activity and provide specific support to individuals with medical conditions or in part-time employment.

Examination of daily-level associations between posttraumatic stress disorder symptoms and COVID-19 worries.

OBJECTIVE: Stressors and worries related to the COVID-19 pandemic have contributed to the onset and exacerbation of psychological symptoms such as posttraumatic stress disorder (PTSD). Using a microlongitudinal framework, we uniquely investigated bidirectional associations between daily-level PTSD symptoms and COVID-19 worries. METHOD: Data from 42 trauma-exposed university students (Mage = 22.67 ± 5.02, 86.7% female) were collected between March and August 2020. Participants completed daily surveys for 10 days to assess PTSD symptom severity and COVID-19 worries. Multilevel regression was conducted to examine both lagged and simultaneous models of daily person-centered mean PTSD symptom severity predicting COVID-19 worries, and vice-versa. RESULTS: Days with greater COVID-19 worries were associated with greater same-day (b = .53, SE = .19, p = .006) and next-day (b = .65, SE = .21, p = .003) PTSD symptom severity. Additionally, days with greater PTSD symptom severity were associated with greater same-day COVID-19 worries (b = .06, SE = .02, p = .006). CONCLUSIONS: COVID-19 worries may influence same-day and next-day PTSD symptoms, and PTSD symptoms may influence same day COVID-19 worries. Findings substantiate the interplay between ongoing stress related to the COVID-19 pandemic and posttrauma symptoms and support therapeutically targeting COVID-19 stress in PTSD treatments to potentially impact posttrauma symptoms. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Implementation of a non-emergent medical transportation programme at an integrated health system.

OBJECTIVES: To implement a unified non-emergency medical transportation (NEMT) service across a large integrated healthcare delivery network. METHODS: We assessed needs among key organisational stakeholders, then reviewed proposals. We selected a single NEMT vendor best aligned with organisational priorities and implemented this solution system-wide. RESULTS: Our vendor's hybrid approach combined rideshares with contracted vehicles able to serve patients with equipment and other needs. After 6195 rides in the first year, we observed shorter wait times and lower costs compared with our prior state. DISCUSSION: Essential lessons included (1) understanding user and patient needs, (2) obtaining complete, accurate and comprehensive baseline data and (3) adapting existing workflows-rather than designing de novo-whenever possible. CONCLUSIONS: Our implementation of a single-vendor NEMT solution validates the need for NEMT at large healthcare organisations, geographical challenges to establishing NEMT organisation-wide, and the importance of baseline data and stakeholder engagement.

Challenges in interpreting cytokine data in COVID-19 affect patient care and management.

Challenges in using cytokine data are limiting Coronavirus Disease 2019 (COVID-19) patient management and comparison among different disease contexts. We suggest mitigation strategies to improve the accuracy of cytokine data, as we learn from experience gained during the COVID-19 pandemic.