CRNA Engagement During the COVID-19 Crisis: Optimization of Resource Management, Organizational Climate, and Contributions to Care.

COVID-19 has a strong hold on New York City, and it has similar effects on other areas of the country. As COVID-19 strains healthcare systems and the certified registered nurse anesthetists (CRNAs) who work within them, optimization of CRNA organizational climate promotes transformation of care delivery and may positively impact provider and patient outcomes. This article describes one healthcare system's newly refined processes for managing the surge of COVID-19 patients. It also describes the novel contributions of CRNAs to all aspects of care provision from supply and resource management to management of patients in critical care environments, to refinement of intubation and airway management for this patient population. Lessons learned during this pandemic from two facilities within this healthcare system are described. When a healthcare system's culture respects and encourages collaboration and innovation, dynamic changes can be implemented effectively during times of crises. Also, critically helpful is when CRNA organizational climate promotes equal partnerships with physicians, and administration values the CRNA contributions to care. In this case, collaboration of all stakeholders promoted best practices and improved care provision. Lessons learned may be applied to facilities where the COVID-19 surge is occurring.

Molecular Analyses of Clinical Isolates and Recombinant SARS-CoV-2 Carrying B.1 and B.1.617.2 Spike Mutations Suggest a Potential Role of Non-Spike Mutations in Infection Kinetics.

Some of the emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants are less susceptible to neutralization with post-vaccine sera and monoclonal antibodies targeting the viral spike glycoprotein. This raises concerns of disease control, transmissibility, and severity. Numerous substitutions have been identified to increase viral fitness within the nucleocapsid and nonstructural proteins, in addition to spike mutations. Therefore, we sought to generate infectious viruses carrying only the variant-specific spike mutations in an identical backbone to evaluate the impact of spike and non-spike mutations in the virus life cycle. We used en passant mutagenesis to generate recombinant viruses carrying spike mutations of B.1 and B.1.617.2 variants using SARS-CoV-2- bacterial artificial chromosome (BAC). Neutralization assays using clinical sera yielded comparable results between recombinant viruses and corresponding clinical isolates. Non-spike mutations for both variants neither seemed to effect neutralization efficiencies with monoclonal antibodies nor the response to treatment with inhibitors. However, live-cell imaging and microscopy revealed differences, such as persisting syncytia and pronounced cytopathic effect formation, as well as their progression between BAC-derived viruses and clinical isolates in human lung epithelial cell lines and primary bronchial epithelial cells. Complementary RNA analyses further suggested a potential role of non-spike mutations in infection kinetics.

Fluorescence-Based Measurements of Membrane-Bound Angiotensin Converting Enzyme 2 Activity Using Xenopus Laevis Oocytes.

Functional investigations of enzymes involving cellular expression systems are important for pharmacological studies. The precise control of expression is challenging in transiently transfected mammalian cell lines. Here, we explored the ability of Xenopus laevis oocytes to express a membrane-bound enzyme for functional characterization using standard 96-well plates and a fluorescence-based plate reader assay. We microinjected oocytes with cRNA encoding the angiotensin converting enzyme 2 (ACE2) and measured the enzymatic activity in single oocytes using a commercial fluorescence-based assay. The injected oocytes showed up to a 50-fold increase in fluorescence compared to uninjected oocytes. This fluorescence intensity was dose-dependent on the amount of ACE2 cRNA. These results suggest that Xenopus oocytes can be used for the functional evaluation of membrane-bound enzymes, decreasing the experimental workload.