Digital Disease Surveillance for Emerging Infectious Diseases: An Early Warning System Using the Internet and Social Media Data for COVID-19 Forecasting in Canada.

BACKGROUND: Emerging Infectious Diseases (EID) are a significant threat to population health globally. We aimed to examine the relationship between internet search engine queries and social media data on COVID-19 and determine if they can predict COVID-19 cases in Canada. METHODS: We analyzed Google Trends (GT) and Twitter data from 1/1/2020 to 3/31/2020 in Canada and used various signal-processing techniques to remove noise from the data. Data on COVID-19 cases was obtained from the COVID-19 Canada Open Data Working Group. We conducted time-lagged cross-correlation analyses and developed the long short-term memory model for forecasting daily COVID-19 cases. RESULTS: Among symptom keywords, "cough," "runny nose," and "anosmia" were strong signals with high cross-correlation coefficients >0.8 ( rCough = 0.825, t - 9; rRunnyNose = 0.816, t - 11; rAnosmia = 0.812, t - 3 ), showing that searching for "cough," "runny nose," and "anosmia" on GT correlated with the incidence of COVID-19 and peaked 9, 11, and 3 days earlier than the incidence peak, respectively. For symptoms- and COVID-related Tweet counts, the cross-correlations of Tweet signals and daily cases were rTweetSymptoms = 0.868, t - 11 and tTweetCOVID = 0.840, t - 10, respectively. The LSTM forecasting model achieved the best performance (MSE = 124.78, R2 = 0.88, adjusted R2 = 0.87) using GT signals with cross-correlation coefficients >0.75. Combining GT and Tweet signals did not improve the model performance. CONCLUSION: Internet search engine queries and social media data can be used as early warning signals for creating a real-time surveillance system for COVID-19 forecasting, but challenges remain in modelling.

Investigating Canadian Public Attitudes Toward COVID-19 Vaccine Mandates with a Nested Analysis Framework.

BACKGROUND: Social media is an important medium for studying public attitudes toward COVID-19 vaccine mandates in Canada, and Reddit network communities are a good source for this. METHODS: This study applied a "nested analysis" framework. We collected 20378 Reddit comments via the Pushshift API and developed a BERT-based binary classification model to screen for relevance to COVID-19 vaccine mandates. We then used a Guided Latent Dirichlet Allocation (LDA) model on relevant comments to extract key topics and assign each comment to its most relevant topic. RESULTS: There were 3179 (15.6%) relevant and 17199 (84.4%) irrelevant comments. Our BERT-based model achieved 91% accuracy trained with 300 Reddit comments after 60 epochs. The Guided LDA model had an optimal coherence score of 0.471 with four topics: travel, government, certification, and institutions. Human evaluation of the Guided LDA model showed an 83% accuracy in assigning samples to their topic groups. CONCLUSION: We develop a screening tool for filtering and analyzing Reddit comments on COVID-19 vaccine mandates through topic modelling. Future research could develop more effective seed word-choosing and evaluation methods to reduce the need for human judgment.

Oral health behaviors for young low-income urban children during the COVID-19 pandemic: a mixed methods analysis (preprint)

This research assessed oral health behaviors changes in urban families with young children during the stay-at-home period of the COVID-19 pandemic. Survey data on oral health behaviors were collected in homes at three points over one year before COVID-19, and then via phone during COVID-19. Multivariate logistic regression was used to model tooth brushing frequency. A subset of parents completed in-depth interviews via video/phone that expanded on oral health and COVID-19. Key informant interviews via video/phone were also conducted with leadership from 20 clinics and social service agencies. Interview data were transcribed and coded, and themes were extracted. COVID-19 data collection went from Nov 2020 – August 2021. Of the 387 parents invited, 254 completed surveys in English or Spanish (65.6%) during COVID-19. Fifteen key informant (25 participants) and 21 parent interviews were conducted. The mean child age was approximately 4.3 years. Children identified as mainly Hispanic (57%) and Black race (38%). Parents reported increased child tooth brushing frequency during the pandemic. Parent interviews highlighted significant changes in family routines that impacted oral health behaviors and eating patterns, suggesting less optimal brushing and nutrition. This was linked to changed home routines and social presentability. Key informants described major disruptions in their oral health services and significant family fear and stress. In conclusion, the stay-at-home period of the COVID-19 pandemic was a time of extreme routine change and stress for families. Oral health interventions that target family routines and social presentability are important for families during times of extreme crisis.

Metatranscriptomics to characterize respiratory virome, microbiome, and host response directly from clinical samples.

We developed a metatranscriptomics method that can simultaneously capture the respiratory virome, microbiome, and host response directly from low biomass samples. Using nasal swab samples, we capture RNA virome with sufficient sequencing depth required to assemble complete genomes. We find a surprisingly high frequency of respiratory syncytial virus (RSV) and coronavirus (CoV) in healthy children, and a high frequency of RSV-A and RSV-B co-detections in children with symptomatic RSV. In addition, we have identified commensal and pathogenic bacteria and fungi at the species level. Functional analysis revealed that H. influenzae was highly active in symptomatic RSV subjects. The host nasal transcriptome reveled upregulation of the innate immune system, anti-viral response and inflammasome pathway, and downregulation of fatty acid pathways in children with symptomatic RSV. Overall, we demonstrate that our method is broadly applicable to infer the transcriptome landscape of an infected system, surveil respiratory infections, and to sequence RNA viruses directly from clinical samples.

Upper respiratory tract microbiota dynamics following COVID-19 in adults.

To date, little is known about the effect of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for the coronavirus disease 2019 (COVID-19) pandemic, on the upper respiratory tract (URT) microbiota over time. To fill this knowledge gap, we used 16S ribosomal RNA gene sequencing to characterize the URT microbiota in 48 adults, including (1) 24 participants with mild-to-moderate COVID-19 who had serial mid-turbinate swabs collected up to 21 days after enrolment and (2) 24 asymptomatic, uninfected controls who had mid-turbinate swabs collected at enrolment only. To compare the URT microbiota between groups in a comprehensive manner, different types of statistical analyses that are frequently employed in microbial ecology were used, including ⍺-diversity, ß-diversity and differential abundance analyses. Final statistical models included age, sex and the presence of at least one comorbidity as covariates. The median age of all participants was 34.00 (interquartile range=28.75-46.50) years. In comparison to samples from controls, those from participants with COVID-19 had a lower observed species index at day 21 (linear regression coefficient=-13.30; 95 % CI=-21.72 to -4.88; q=0.02). In addition, the Jaccard index was significantly different between samples from participants with COVID-19 and those from controls at all study time points (PERMANOVA q<0.05 for all comparisons). The abundance of three amplicon sequence variants (ASVs) (one Corynebacterium ASV, Frederiksenia canicola, and one Lactobacillus ASV) were decreased in samples from participants with COVID-19 at all seven study time points, whereas the abundance of one ASV (from the family Neisseriaceae) was increased in samples from participants with COVID-19 at five (71.43 %) of the seven study time points. Our results suggest that mild-to-moderate COVID-19 can lead to alterations of the URT microbiota that persist for several weeks after the initial infection.

Evaluating the Women's Empowerment and Leadership Initiative: Supporting mentorship, career satisfaction, and well-being among pediatric anesthesiologists.

The Society for Pediatric Anesthesia launched the Women's Empowerment and Leadership Initiative (WELI) in 2018 to empower highly productive women pediatric anesthesiologists to achieve equity, promotion, and leadership. WELI is focused on six career development domains: promotion and leadership, networking, conceptualization and completion of projects, mentoring, career satisfaction, and sense of well-being. We sought feedback about whether WELI supported members' career development by surveys emailed in November 2020 (baseline), May 2021 (6 months), and January 2022 (14 months). Program feedback was quantitatively evaluated by the Likert scale questions and qualitatively evaluated by extracting themes from free-text question responses. The response rates were 60.5% (92 of 152) for the baseline, 51% (82 of 161) for the 6-month, and 52% (96 of 185) for the 14-month surveys. Five main themes were identified from the free-text responses in the 6- and 14-month surveys. Members reported that WELI helped them create meaningful connections through networking, obtain new career opportunities, find tools and projects that supported their career advancement and promotion, build the confidence to try new things beyond their comfort zone, and achieve better work-life integration. Frustration with the inability to connect in-person during the coronavirus-19 pandemic was highlighted. Advisors further stated that WELI helped them improve their mentorship skills and gave them insight into early career faculty issues. Relative to the baseline survey, protégés reported greater contributions from WELI at 6 months in helping them clarify their priorities, increase their sense of achievement, and get promoted. These benefits persisted through 14 months. Advisors reported a steady increase in forming new meaningful relationships and finding new collaborators through WELI over time. All the members reported that their self-rated mentoring abilities improved at 6 months with sustained improvement at 14 months. Thus, programs such as WELI can assist women anesthesiologists and foster gender equity in career development, promotion, and leadership.

Sequence-specific capture and concentration of viral RNA by type III CRISPR system enhances diagnostic.

Type-III CRISPR-Cas systems have recently been adopted for sequence-specific detection of SARS-CoV-2. Here, we repurpose the type III-A CRISPR complex from Thermus thermophilus (TtCsm) for programmable capture and concentration of specific RNAs from complex mixtures. The target bound TtCsm complex generates two cyclic oligoadenylates (i.e., cA3 and cA4) that allosterically activate ancillary nucleases. We show that both Can1 and Can2 nucleases cleave single-stranded RNA, single-stranded DNA, and double-stranded DNA in the presence of cA4. We integrate the Can2 nuclease with type III-A RNA capture and concentration for direct detection of SARS-CoV-2 RNA in nasopharyngeal swabs with 15 fM sensitivity. Collectively, this work demonstrates how type-III CRISPR-based RNA capture and concentration simultaneously increases sensitivity, limits time to result, lowers cost of the assay, eliminates solvents used for RNA extraction, and reduces sample handling.

The First COVID-19 Pandemic Wave and the Effect on Health Care Trainees: A National Survey Study.

BACKGROUND: This study observes the trends and patterns among trainees during the coronavirus disease 2019 (COVID-19) pandemic and their response to resident education and hospital/program support. METHODS: An anonymous online 31-question survey was distributed to medical students and postgraduate year residents. Topics included were demographics, clinical responsibilities, educational/curricula changes, and trainee wellness. Descriptive analysis was performed for each set of demographic groupings as well as 2 and 3 group comparisons. RESULTS: Total 1051 surveys collected, 930 used for analysis: 373 (40.1%) male, 434 (46.6%) aged 30-34 years, 588 (63.2%) white, 417 (44.8%) married, 168 (18%) with children, and 323 (34.7%) from the Northeast region. The Northeast experienced difficulty sleeping, feelings of guilt, hopelessness, and changes in appetite (P = .0077). The pandemic interfered significantly with relationships and living situations (P < .0001). Trainees 18-34 years believed the pandemic affected residency training (P < .0001). Surgical residents were concerned about reaching numbers of operative procedures to graduate (P < .0001). Residency programs adhered to ACGME work restrictions (P < .0001). CONCLUSION: We aim to provide continued educational support for our trainees' clinical development and well-being during the COVID-19 pandemic.

Development and performance of a point-of-care rapid antigen test for detection of SARS-COV-2 variants.

Background: SARS-CoV-2 antigen-based tests are well-calibrated to infectiousness and have a critical role to play in the COVID-19 public health response. We report the development and performance of a unique lateral flow immunoassay (LFA). Methods: Combinations of several monoclonal antibodies targeting multiple antigenic sites on the SARS-CoV-2 nucleocapsid protein (NP) were isolated, evaluated, and chosen for the development of a LFA termed CoV-SCAN (BioMedomics, Inc.). Clinical point-of-care studies in symptomatic and asymptomatic individuals were conducted to evaluate positive predictive agreement (PPA) and negative predictive agreement (NPA) with RT-PCR as comparator. Results: In laboratory testing, CoV-SCAN detected 14 recombinant N-proteins of SARS-CoV-2 variants with sensitivity in the range of 0.2-3.2 ng/mL, and 10 authentic SARS-CoV-2 variants with sensitivity in the range of 1.6-12.5 TCID50/swab. No cross reactivity was observed with other human coronaviruses or other respiratory pathogens. In clinical point-of-care testing on 148 individuals over age 2 with symptoms of ≤5 days, PPA was 87.2% (CI 95: 78.3-94.8%) and NPA was 100% (CI 95: 94.2-100%). In another 884 asymptomatic individuals, PPA was 85.7% (CI 95: 42.1-99.6%) and 99.7% (99.0-99.9%). Overall, CoV-SCAN detected over 97.2% of specimens with CT values <30 and 93.8% of nasal swab specimens with the Omicron variant, even within the first 2 days after symptom onset. Conclusions: The unique construction of CoV-SCAN using two pairs of monoclonal antibodies has resulted in a test with high performance that remains durable across multiple variants in both laboratory and clinical evaluations. CoV-SCAN should identify almost all individuals harboring infectious SARS-CoV-2. Summary: Unique construction of a point-of-care rapid antigen test using two pairs of monoclonal antibodies has led to good performance that remained durable across multiple variants in laboratory and clinical evaluations. Test should identify almost all individuals harboring infectious SARS-CoV-2.

Service process factors affecting patients' and clinicians' experiences on rapid teleconsultation implementation in out-patient neurology services during COVID-19 pandemic: a scoping review.

BACKGROUND: The adoption of teleconsultation for outpatient neurology services was limited until the onset of the COVID-19 pandemic which forced many outpatient neurology services to rapidly switch to virtual models. However, it remains unclear how this change has impacted patients' and clinicians' perceptions of service quality. The purpose of this scoping review is to identify process factors that influence patients' and clinicians' experiences of outpatient teleconsultation services during COVID-19. METHODS: Arksey and O'Malley scoping review framework was used to search PubMed, Scopus, CINAHL, and PsycInfo for original peer-reviewed research studies that examined the experiences of synchronous teleconsultation between a clinician and patient in a home-setting since the World Health Organization announced the COVID-19 global pandemic. The service quality model SERVQUAL was used to conduct a deductive thematic analysis to identify the key factors that impacted the patients' and clinicians' perception of teleconsultation services. RESULTS: A total of nineteen studies published between January 1, 2020, and April 17, 2021, were identified. The most common service process factors affecting the patients' and clinicians' experiences of teleconsultation were technical issues, addressing logistical needs, communication, ability to perform clinical activities, appropriate triage, and administrative support. CONCLUSIONS: Our findings identified six key service process factors affecting the patients' and clinicians' teleconsultation experiences in outpatient neurology services. The need for improvement of triage process and standardizing administrative virtual care pathway are identified as important steps to improve patients and clinicians' teleconsultation experiences compared to pre-COVID era. More research is needed to assess outpatient neurology teleconsultation service quality from patients' and clinicians' perspectives.

The Intention to Use Online Groceries Shopping during the COVID19 Pandemic

The Coronavirus disease (COVID19) has brought many impacts to the activities including changed the habit for shopping the daily groceries. The purpose of this research is to examine the intention to use the Online Groceries Shopping (OGS) based on the framework of Technology Acceptance Model (TAM) with two primary factors that influence the intention to use technology OGS: perceived usefulness and perceived ease of use. This is a quantitative research with non-probability sampling techniques of 317 respondents measured by 5 Point Likert scale and using Structural Equation Model (SEM) Smart PLS version 3 to analyze the data. The results obtained show all variables have positive and significant relationship to the intention to use OGS during the Covid19 Pandemic. © 2021 IEEE.

Severe COVID-19 Is Associated With an Altered Upper Respiratory Tract Microbiome.

Background: The upper respiratory tract (URT) is the portal of entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and SARS-CoV-2 likely interacts with the URT microbiome. However, understanding of the associations between the URT microbiome and the severity of coronavirus disease 2019 (COVID-19) is still limited. Objective: Our primary objective was to identify URT microbiome signature/s that consistently changed over a spectrum of COVID-19 severity. Methods: Using data from 103 adult participants from two cities in the United States, we compared the bacterial load and the URT microbiome between five groups: 20 asymptomatic SARS-CoV-2-negative participants, 27 participants with mild COVID-19, 28 participants with moderate COVID-19, 15 hospitalized patients with severe COVID-19, and 13 hospitalized patients in the ICU with very severe COVID-19. Results: URT bacterial load, bacterial richness, and within-group microbiome composition dissimilarity consistently increased as COVID-19 severity increased, while the relative abundance of an amplicon sequence variant (ASV), Corynebacterium_unclassified.ASV0002, consistently decreased as COVID-19 severity increased. Conclusions: We observed that the URT microbiome composition significantly changed as COVID-19 severity increased. The URT microbiome could potentially predict which patients may be more likely to progress to severe disease or be modified to decrease severity. However, further research in additional longitudinal cohorts is needed to better understand how the microbiome affects COVID-19 severity.

GP perceptions of telehealth services in Australia: a qualitative study.

BACKGROUND: Primary care providers have been rapidly transitioning from in-person to telehealth care during the 2019 coronavirus (COVID-19) pandemic. There is an opportunity for new research in a rapidly evolving area, where evidence for telehealth services in primary care in the Australian setting remains limited. AIM: To explore general practitioner (GP) perceptions on providing telehealth (telephone and video consultation) services in primary care in Australia. DESIGN & SETTING: A qualitative study using semi-structured interviews to gain an understanding of GP perceptions on telehealth use in Australia. METHOD: GPs across Australia were purposively sampled. Semi-structured interviews were conducted, recorded, and transcribed verbatim for analysis. Transcripts were analysed using inductive thematic analysis to identify initial codes, which were then organised into themes. RESULTS: Fourteen GPs were interviewed. Two major themes that described GP perceptions of telehealth were: (1) existence of business and financial pressures in general practice; and (2) providing quality of care in Australia. These two themes interacted with four minor themes: (3) consumer-led care; (4) COVID-19 as a driver for telehealth reimbursement and adoption; (5) refining logistical processes; and (6) GP experiences shape telehealth use. CONCLUSION: This study found that multiple considerations influenced GP choice of in-person, videoconference, or telephone consultation mode. For telehealth to be used routinely within primary care settings, evidence that supports the delivery of higher quality care to patients through telehealth and sustainable funding models will be required.

Empowering Women as Leaders in Pediatric Anesthesiology: Methodology, Lessons, and Early Outcomes of a National Initiative.

Research has shown that women have leadership ability equal to or better than that of their male counterparts, yet proportionally fewer women than men achieve leadership positions and promotion in medicine. The Women's Empowerment and Leadership Initiative (WELI) was founded within the Society for Pediatric Anesthesia (SPA) in 2018 as a multidimensional program to help address the significant career development, leadership, and promotion gender gap between men and women in anesthesiology. Herein, we describe WELI's development and implementation with an early assessment of effectiveness at 2 years. Members received an anonymous, voluntary survey by e-mail to assess whether they believed WELI was beneficial in several broad domains: career development, networking, project implementation and completion, goal setting, mentorship, well-being, and promotion and leadership. The response rate was 60.5% (92 of 152). The majority ranked several aspects of WELI to be very or extremely valuable, including the protégé-advisor dyads, workshops, nomination to join WELI, and virtual facilitated networking. For most members, WELI helped to improve optimism about their professional future. Most also reported that WELI somewhat or absolutely contributed to project improvement or completion, finding new collaborators, and obtaining invitations to be visiting speakers. Among those who applied for promotion or leadership positions, 51% found WELI to be somewhat or absolutely valuable to their application process, and 42% found the same in applying for leadership positions. Qualitative analysis of free-text survey responses identified 5 main themes: (1) feelings of empowerment and confidence, (2) acquisition of new skills in mentoring, coaching, career development, and project implementation, (3) clarification and focus on goal setting, (4) creating meaningful connections through networking, and (5) challenges from coronavirus disease 2019 (COVID-19) and the inability to sustain the advisor-protégé connection. We conclude that after 2 years, the WELI program has successfully supported career development for the majority of protégés and advisors. Continued assessment of whether WELI can meaningfully contribute to attainment of promotion and leadership positions will require study across a longer period. WELI could serve as a programmatic example to support women's career development in other subspecialties.

Service Process Factors Affecting Patients’ and Clinicians’ Experiences on Rapid Teleconsultation Implementation in Out-patient Neurology Services during COVID-19 Pandemic: A Scoping Review (preprint)

BackgroundPrior to the COVID-19 outbreak, the adoption of teleconsultation for outpatient neurology services was limited. Since the COVID-19 pandemic, many outpatient neurology services have rapidly switched to teleconsultation worldwide. However, it remains unclear how this change has impacted patients' and clinicians' perceptions of service quality. The purpose of this scoping review is to identify process factors that influence patients' and clinicians' experiences of outpatient teleconsultation services during COVID-19.MethodsArksey and O’Malley scoping review framework was used to search PubMed, Scopus, CINAHL, and PsycInfo for original peer-reviewed research studies that examined the experiences of synchronous teleconsultation between a clinician and patient in a home-setting since the World Health organization announced the COVID-19 global pandemic. The service quality model SERVQUAL was used to conduct a deductive thematic analysis to identify the key factors that impacted the patients’ and clinicians’’ perception of teleconsultation services. Results A total of nineteen studies published between January 1, 2020, and April 17, 2021, were identified. The most common service process factors affecting the patients' and clinicians' experiences of teleconsultation were technical issues, addressing logistical needs, communication, ability to perform clinical activities, appropriate triage, and administrative support. Conclusions Our findings identified six key service process factors affecting the patients' and clinicians' teleconsultation experiences in outpatient neurology services. The need for a new triage system and standardizing administrative virtual care workflow are identified as important steps to improve patients and clinicians’ teleconsultation experiences compared to pre-COVID. More research is needed to assess outpatient neurology teleconsultation service quality from patients' and clinicians' perspectives.

Non-pharmaceutical interventions and COVID-19 cases in US summer camps: results from an American Camp Association survey.

BACKGROUND: Most camps remained closed during Summer 2020, due to concerns regarding child transmission of SARS-CoV-2 and limited information about the effectiveness of non-pharmaceutical interventions (NPIs) within child congregate settings. METHODS: We surveyed US camps about on-site operations, camper and staff demographics, COVID-19 cases among campers and staff, and NPI usage as related to pre-camp quarantine, facial coverings, physical distancing, cleaning and facility modifications. For all NPIs, save quarantine, responses were provided on a 5-point Likert scale format. RESULTS: Within 486 on-site camps, a range of NPIs were instituted, most often related to reduced camper interactions, staff face coverings, cleaning and hand hygiene. Camper facial coverings were less common, with campers always wearing masks at ~34% of the camps. Approximately 15% of camps reported 1+ confirmed COVID-19 case in either campers or staff, with three camps reporting a COVID-19 outbreak. In both single and multi-NPI analyses, the risk of COVID-19 cases was lowest when campers always wore facial coverings. Constant use of staff facial coverings and targeted physical distancing measures, but not pre-camp quarantine, also reduced COVID-19 risks. CONCLUSIONS: We found constant facial coverings, especially for campers, and targeted physical distancing measures to reduce risks of SARS-CoV-2 transmission within summer camps. Our findings provide valuable insights for future operations of summer camps and other child congregate settings regarding the use of NPIs to reduce the risk of SARS-CoV-2 infection.

Evaluation of sample pooling using the SAMBA II SARS-CoV-2 test.

BACKGROUND: Screening of infectious asymptomatic or pre-symptomatic individuals for SARS-CoV-2 is at present a key to controling the COVID-19 pandemic. In order to expand testing capability and limit cost, pool testing of asymtomatic individuals has been proposed, provided assay performance is not significantly affected. METHODS: Combined nose and throat (N/T) swabs collected from COVID-19 infected or non-infected individuals were tested using SAMBA II individually and in pools of four (one positive and 3 negative). The evaluation was conducted by the manufacturer and an independent NHS site. Ct cycles of individual positives and pooled positives were determined by qRT-PCR. RESULTS: In 42 pools containing a single positive sample with Ct values ranging between 17 and 36, 41 pools (97.6 %) were found positive by the SARS-CoV-2 SAMBA II test. The false-negative pool by SAMBA was also negative by both reference methods used in this evaluation.The individual positive sample in this pool was positive by SAMBA (Orf only) and by one of the reference methods (S gene only, Ct 35) but negative by the second reference method indicating that the sample itself was very low viral load. All 78 pools containing 4 negative swabs were negative (100 % specificity). DISCUSSION: The preliminary data of the evaluation indicated a high level of performance in both sensitivity and specificity of the SAMBA II assay when used to test pools of 4 patient samples. The implementation of this pooled protocol can increase throughput and reduce cost/test when the prevalence of COVID is low.

Conferences in the time of COVID: notes on organizing and delivering the first Brain Conference.

To further fulfil their missions of promoting teaching, education and research in neurology and related clinical-academic disciplines, the Guarantors of Brain and the Brain journal family invited delegates to the first Brain Conference in Spring of this year. This event aimed to deliver excellent teaching and scientific presentations across a broad spectrum of neuroscience fields, with the key aim of making the content as accessible as possible. We hoped to capitalize on the benefits of an online format, whilst trying to capture a little of the joy of the in-person meeting. This article reports on the approach and practical choices made to achieve these goals, and we hope this will provide some guidance and advice to others organizing their own online conference.

CD73 Maintains Hepatocyte Metabolic Integrity and Mouse Liver Homeostasis in a Sex-Dependent Manner.

BACKGROUND & AIMS: Metabolic imbalance and inflammation are common features of chronic liver diseases. Molecular factors controlling these mechanisms represent potential therapeutic targets. CD73 is the major enzyme that dephosphorylates extracellular adenosine monophosphate (AMP) to form the anti-inflammatory adenosine. CD73 is expressed on pericentral hepatocytes, which are important for long-term liver homeostasis. We aimed to determine if CD73 has nonredundant hepatoprotective functions. METHODS: Liver-specific CD73 knockout (CD73-LKO) mice were generated by targeting the Nt5e gene in hepatocytes. The CD73-LKO mice and hepatocytes were characterized using multiple approaches. RESULTS: Deletion of hepatocyte Nt5e resulted in an approximately 70% reduction in total liver CD73 protein (P < .0001). Male and female CD73-LKO mice developed normally during the first 21 weeks without significant liver phenotypes. Between 21 and 42 weeks, the CD73-LKO mice developed spontaneous-onset liver disease, with significant severity in male mice. Middle-aged male CD73-LKO mice showed hepatocyte swelling and ballooning (P < .05), inflammation (P < .01), and variable steatosis. Female CD73-LKO mice had lower serum albumin levels (P < .05) and increased inflammatory genes (P < .01), but did not show the spectrum of histopathologic changes in male mice, potentially owing to compensatory induction of adenosine receptors. Serum analysis and proteomic profiling of hepatocytes from male CD73-LKO mice showed significant metabolic imbalance, with increased blood urea nitrogen (P < .0001) and impairments in major metabolic pathways, including oxidative phosphorylation and AMP-activated protein kinase (AMPK) signaling. There was significant hypophosphorylation of AMPK substrates in CD73-LKO livers (P < .0001), while in isolated hepatocytes treated with AMP, soluble CD73 induced AMPK activation (P < .001). CONCLUSIONS: Hepatocyte CD73 supports long-term metabolic liver homeostasis through AMPK in a sex-dependent manner. These findings have implications for human liver diseases marked by CD73 dysregulation.

Intrinsic signal amplification by type III CRISPR-Cas systems provides a sequence-specific SARS-CoV-2 diagnostic.

There is an urgent need for inexpensive new technologies that enable fast, reliable, and scalable detection of viruses. Here, we repurpose the type III CRISPR-Cas system for sensitive and sequence-specific detection of SARS-CoV-2. RNA recognition by the type III CRISPR complex triggers Cas10-mediated polymerase activity, which simultaneously generates pyrophosphates, protons, and cyclic oligonucleotides. We show that all three Cas10-polymerase products are detectable using colorimetric or fluorometric readouts. We design ten guide RNAs that target conserved regions of SARS-CoV-2 genomes. Multiplexing improves the sensitivity of amplification-free RNA detection from 107 copies/µL for a single guide RNA to 106 copies/µL for ten guides. To decrease the limit of detection to levels that are clinically relevant, we developed a two-pot reaction consisting of RT-LAMP followed by T7-transcription and type III CRISPR-based detection. The two-pot reaction has a sensitivity of 200 copies/µL and is completed using patient samples in less than 30 min.