ABSTRACT
Viral RNA in fine (< 5 µm) aerosols from 13 patients infected by the SARS-CoV-2 virus were obtained using the Gesundheit-II (G-II) equipment which collects respiratory emissions. The collection was performed in isolation wards of the National Centre for Infectious Diseases of Singapore under an approved protocol. The patients breathed normally for 30 minutes, talk, and sing for 15 minutes each (with 30 minutes rest in between activity) into a specially designed aerosol collector in two size fractions. The coarse fraction (> 5 µm) and the fine aerosols (< 5 µm) are subsequently collected and subjected to PCR analysis for their viral load quantification. Viral RNA detected from 59% of the patients showed that patients earlier in illness were more likely to emit detectable RNA, and loads differed significantly between breathing, talking, and singing. © 2022 17th International Conference on Indoor Air Quality and Climate, INDOOR AIR 2022. All rights reserved.
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Introduction: The outbreak of coronavirus disease (COVID-19) in December 2019 called for a rapid solution, leading to repurposing of existing drugs. Due to its immunomodulatory effect and antiviral properties, hydroxychloroquine (HCQ) has been used in early 2020 for treatment of COVID-19 patients. This study was conducted to evaluate the treatment outcome of HCQ monotherapy in Malaysia. Methods: A retrospective cohort study was conducted in COVID-19 ward in Hospital Kuala Lumpur (HKL), from March to April 2020. A total of 446 COVID-19 patients were recruited, only 325 patients were finally included for analysis. Statistical analysis was done using SPSS, with a significant value set at p<0.05. Results: The mean age of the patients were 38.5 ±15.5. They were majority male, (n=210, 64.6%) Malaysian (n=239, 73.5%) and Malay ethnicity (n=204, 62.8%). Ninety-one (28%) patients received HCQ monotherapy. HCQ monotherapy was associated with worse outcome (OR: 10.29, 95% CI 1.17-90.80). There was a significant difference in mean length of stay between those with and without HCQ treatment (t323=5.868, p<0.001, 95% CI, 2.56-5.31). The average length of stay for HCQ treated group was 3.84 days longer than those without treatment. 6.6% of the patient receiving HCQ monotherapy encountered adverse drug effects. Conclusion: Similar to study reported worldwide, our study demonstrated that HCQ did not improve length of stay and the outcome of COVID-19 patients. © 2023 Authors. All rights reserved.
ABSTRACT
The emergence of the novel severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) Coronavirus resulted in a global pandemic due to its nature of rapid transmission and variable severities that facilitated its spread worldwide. Correspondingly, owing to advances in molecular technologies, information on this virus is generated at an unprecedented pace. Since the onset of the pandemic, multiple highthroughput "omics" analyses - including transcriptomics and proteomics of different viral infection models - have been made readily available to the research and wider community. The availability and ability to rapidly generate these data facilitate the deciphering of virus–host interactions during SARS-CoV-2 infection - thus enhancing understanding of the viral transmission, host susceptibility, pathogenesis, viral evolution, and disease complications. Such information is vital for eventual applications towards biomarker and treatment discovery against Coronavirus disease 2019 (COVID-19), and can serve as useful models for future pandemic responses. © 2023 by World Scientific Publishing Co. Pte. Ltd.
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Background: We aim to explore the factors related to job satisfaction among pain physicians and identify the reasons why individuals minimize or stop practicing outpatient pain medicine.Objectives/Study Design: This is a survey-based study with the primary goal to identify factors determining job satisfaction and dissatisfaction among pain medicine fellowship graduates who continue to practice and those who are no longer practicing interventional pain. A secondary goal is to elucidate reasons for anesthesiologists trained in pain medicine to leave pain medicine, despite an additional year of training, and to work as general anesthesiologists.Methods: In this study, all 114 pain program directors listed on the Accreditation Council for Graduate Medical Education (ACGME) website, or their administrative assistants were directly contacted via email. All email addresses were obtained from the ACGME website. The survey opened in September 2021, with reminder emails sent before the closing of the survey in December 2021. A final reminder email was sent 4 weeks prior to the closing of the survey.Results: Of all the respondents, 79 (89.77%) were currently practicing pain medicine, and 9 (10.23%) were no longer practicing pain medicine.Limitations: Our study has a major limitation as we are unable to determine the response rate and are limited in the data points gathered.Conclusion: We hope this study will allow for pain medicine fellowship program directors to improve recruitment and retention of pain fellows in the field while addressing the pros and cons of future career aspirations with anesthesiology residents prior to fellowship selection. A larger, more thorough study with an exact response rate can compare the various outcomes based upon different types of settings, such as private practice, partnership, and academia, as well as geographical locations.
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The dynamics of SARS-CoV-2 infection in COVID-19 patients are highly variable, with a subset of patients demonstrating prolonged virus shedding, which poses a significant challenge for disease management and transmission control. In this study, the long-term dynamics of SARS-CoV-2 infection were investigated using a human well-differentiated nasal epithelial cell (NEC) model of infection. NECs were observed to release SARS-CoV-2 virus onto the apical surface for up to 28 days post-infection (dpi), further corroborated by viral antigen staining. Single-cell transcriptome sequencing (sc-seq) was utilized to explore the host response from infected NECs after short-term (3-dpi) and long-term (28-dpi) infection. We identified a unique population of cells harboring high viral loads present at both 3 and 28 dpi, characterized by expression of cell stress-related genes DDIT3 and ATF3 and enriched for genes involved in tumor necrosis factor alpha (TNF-alpha) signaling and apoptosis. Remarkably, this sc-seq analysis revealed an antiviral gene signature within all NEC cell types even at 28 dpi. We demonstrate increased replication of basal cells, absence of widespread cell death within the epithelial monolayer, and the ability of SARS-CoV-2 to replicate despite a continuous interferon response as factors likely contributing to SARS-CoV-2 persistence. This study provides a model system for development of therapeutics aimed at improving viral clearance in immunocompromised patients and implies a crucial role for immune cells in mediating viral clearance from infected epithelia. IMPORTANCE Increasing medical attention has been drawn to the persistence of symptoms (long-COVID syndrome) or live virus shedding from subsets of COVID-19 patients weeks to months after the initial onset of symptoms. In vitro approaches to model viral or symptom persistence are needed to fully dissect the complex and likely varied mechanisms underlying these clinical observations. We show that in vitro differentiated human NECs are persistently infected with SARS-CoV-2 for up to 28 dpi. This viral replication occurred despite the presence of an antiviral gene signature across all NEC cell types even at 28 dpi. This indicates that epithelial cell intrinsic antiviral responses are insufficient for the clearance of SARS-CoV-Z implying an essential role for tissue-resident and infiltrating immune cells for eventual viral clearance from infected airway tissue in COVID-19 patients.
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Background: The highly specialised proximal tubule (PT) nephron segment is responsible for most kidney functions in mammals and is acutely vulnerable to disease, making it a key objective for toxicity screening and disease research. While induced pluripotent stem cell (iPSC)-derived kidney organoids represent a promising approach, the PT remains immature with limited evidence of functional transporters. Here we report the development of PT-enhanced organoids with nephron functionality, enabling improved modelling of PT-relevant conditions including drug-induced toxicity and infection such as SARS-CoV-2. Methods: Standard and fluorescent reporter iPSC lines were subjected to prolonged monolayer differentiations (modified from: Howden et al. EMBO Rep 2019;Vanslambrouck et al. JASN 2019) and precisely-timed morphogens for targets such as WNT, BMP and NOTCH pathways prior to organoid generation (Takasato et al,. Nat Protoc 2016). Maturation was analysed via immunofluorescence, live confocal imaging of fluorescent reporters, transcriptional profiling, transporter function assays, and SARSCoV-2 infectivity. Results: Prolonged nephron progenitor differentiation with simultaneous prevention of spontaneous nephrogenesis resulted in PT-enhanced kidney organoids with elongated and aligned nephrons. Striking proximo-distal nephron orientation resulted from localised WNT antagonism. Improved upregulation of PT-specific markers compared to standard organoids was strengthened by evidence of transporter functionality, including uptake of albumin, organic cations, and cisplatin (eliciting appropriate KIM1 upregulation). This approach also improved expression of SARS-CoV-2 entry factors, confirmed by susceptibility to infection and viral replication. Conclusions: We describe enhanced kidney organoids with improved PT maturity arising from alterations to early mesodermal patterning and delayed nephron initiation. The enhanced conditions also provided more stringent control over nephron spatial arrangement. PT-enhanced organoids provide an ideal model to better understand human PT maturation, inherited and acquired PT disease, and drug toxicity implications.