EXPLORING THE COVID-19 PANDEMIC EXPERIENCE OF MATERNITY CLINICIANS IN A HIGH MIGRANT POPULATION AND LOW COVID PREVALENCE COUNTRY

Background: Australia experienced a low prevalence of COVID-19 in 2020 compared to many other countries. However, maternity care has been impacted with hospital policy driven changes in practice. Little qualitative research has investigated maternity clinicians' perception of the impact of COVID-19 in a high-migrant population. We investigate maternity clinicians' perceptions of patient experience, service delivery and personal experience in a high-migrant population. Methods We conducted semi-structured in-depth interviews with 14 maternity care clinicians in Sydney, New South Wales, Australia. Interviews were conducted from November to December 2020. A reflexive thematic approach was used for data analysis Results: A key theme in the data was 'COVID-19 related travel restrictions result in loss of valued family support for migrant families. However, partners were often 'stepping-up' into the role of missing overseas relatives. The main theme in clinical care was a shift in healthcare delivery away from optimising patient care to a focus on preservation and safety of health staff. Conclusions: Clinicians were of the view migrant women were deeply affected by the loss of traditional support. However, the benefit may be the potential for greater gender equity and bonding opportunities for partners. Conflict with professional beneficence principles and values may result in bending rules when a disconnect exists between relaxed community health orders and restrictive hospital protocols during different phases of a pandemic. This research adds to the literature that migrant women require individualised culturally safe care because of the ongoing impact of loss of support during the COVID-19 pandemic.

ROLE of ACTIN REGULATORS in HIV-1, IAV, & SARS-CoV-2 VIRUS-LIKE PARTICLES PRODUCTION

Background: Human immunodeficiency virus (HIV) and Influenza A virus (IAV) remain a global health concern. Further, emergence of novel coronavirus SARS-CoV-2, which rapidly became global pandemic, increases the concern in biomedical research field for antiviral treatment. To develop new antiviral therapy, we must need to understand the molecular and cellular mechanisms involved in assembly and replication. It is known for some viruses (HIV and IAV) that the host actin cytoskeleton has been involved in various stages of the virus life cycle. Regulation of actin cytoskeleton requires several actin binding proteins, which organize the actin filaments (F-actin) into higher order structures such as actin bundles, branches, filopodia and microvilli, for further assistance in viral particle production. Thus, our objective for this work is to understand the role of these actin regulator proteins, like cofilin and one of its cofactor WDR1, in viral particle assembly and release. Methods: Here we used a combination of different experimental methods like RNA interference, immunoblot, immunoprecipitation, immunofluorescence coupled to confocal and STED fluorescence microscopy. In order to study only virus release, and bypass viral entry, we set up a minimal system for virus-like particles production in transfected cells, giving HIV-1 Gag-VLP, Influenza M1-VLP and SARS-CoV-2 MNE-VLP (developed by D. Muriaux lab). For image analysis, we used Image J software. Statistical analysis was performed with non-parametric t-tests or one-way Anova test. Results: Using siRNA strategy, we have shown that upon knock down of actin protein cofilin or WDR1, HIV-1 and IAV particles production increases in contrario to SARS-CoV-2 VLP release. Further, using immunoprecipitation, we report that HIV-1 Gag is able to form an intracellular complex with WDR1 and cofilin. Similarly, IAV-M1, which like HIV Gag-MA binds with plasma membrane phospholipids, is able to form an intracellular complex with cofilin. These results suggested that virus budding from the host cell plasma membrane seemed restricted by the cofilin/WDR1 complex. Finally, using confocal/STED microscopy on cell producing VLP, we observed actin fibers rearrangement with cell protrusions, suggesting a role for actin in viral particles assembly and release. Conclusion: In conclusion, regulators of actin dynamic are involved in HIV-1 Gag, IAV-M1 and SARS-CoV-2 VLP production but play a differential role in assembly and release of these RNA enveloped viruses.

Allelic variation in class I HLA determines CD8(+) T cell repertoire shape and cross-reactive memory responses to SARS-CoV-2

[Figure: see text].

ABSENCE OF COVID-19 VIRUS WITHIN AN ACTIVE IVF LABORATORY USING STRICT PATIENT SCREENING AND SAFETY CRITERIA

Objective: In the early stages of the COVID-19 pandemic, most IVF clinics stopped the majority of patient treatment cycles to minimize the risk of disease transmission. When ASRM and other professional societies recommended resumption of treatments, procedures were put into place to ensure patient and staff safety. However, the risk of SARS-CoV-2 viral exposure and potential cross contamination within the IVF laboratory remains largely unclear. The objective of this study was to assess the true risk of exposure to SARS-CoV-2 in an active IVF laboratory when strict patient screening procedures are in place. Design: Prospective analysis. Materials and Methods: Prior to restarting IVF treatments, a COVID-19 safety protocol was implemented. Patients and staff were required to wear masks, fill out a symptom-based questionnaire daily, have their temperature taken, and practice social distancing in patient waiting areas. Each female patient undergoing transvaginal oocyte retrieval (TVOR) was required to have a negative SARS-CoV-2 RNA test 3-4 days prior to the procedure. Male partners were not tested. All cases examined utilized ICSI. The first tube of follicular fluid aspirated during TVOR (FF), culture media drops following removal of embryos on day 5 (M), and vitrification solution (VS) after blastocyst cryopreservation were analyzed. Self-inactivating replication incompetent lentivirus particles containing the single stranded viral RNA genome were immediately inoculated into each sample after collection as a positive control for viral RNA stability, prior to direct RNA isolation (M, VS) or sample concentration (FF). For FF, cell debris was removed by centrifugation and filtration (0.22 um) prior to concentration of virus particles with an Amicon filter. RNA was isolated using the optimized QIAamp viral RNA minikit, RNA quantity and quality determined, and cDNA synthesized using SuperScript IV VILO master mix. A multiplex TaqMan-based qPCR assay was developed for SARS-CoV-2 and lentivirus RNA (detection limit 5 SARS-CoV-2 copies/qPCR reaction and 50 viral copies/2 mL sample), and used to test all diagnostic samples. SARS-Cov2 synthetic RNA and lentivirus RNA were used as an RT-qPCR positive control. Samples with no amplification of lentivirus genome were removed from the analysis (false negative). Results: In total, culture medium from 30 patients, vitrification solution from 98 patients, and follicular fluid from 156 patients were analyzed. All samples were negative for the presence of SARS-CoV-2 viral RNA. Conclusions: With stringent safety protocols in place, including patient testing and use of ICSI, the presence of SARS-CoV-2 RNA can be avoided in the IVF laboratory. Importantly, this study does not indicate that virus from an actively infected patient cannot be found in follicular fluid or make its way into the IVF lab. However, it does provide reassurance that with proper patient testing and safety measures, cross-contamination of the virus between gametes and embryos (including within liquid nitrogen storage dewars), as well as exposure of embryologists, is minimal.