Protocol for primary human lung organoid-derived air-liquid interface in vitro model to study response to SARS-CoV-2 (preprint)

This article presents a comprehensive protocol for establishing primary human lung organoid-derived air-liquid interface (ALI) cultures from cryopreserved human lung tissue. These cultures serve as a physiologically relevant model to study human airway epithelium in vitro. The protocol encompasses lung tissue cryostorage, tissue dissociation, lung epithelial organoid generation, and ALI culture differentiation. It also demonstrates SARS-CoV-2 infection in these cultures as an example of their utility. Quality control steps, ALI characterization, and technical readouts for monitoring virus response are included in the study. For additional details on the use and execution of this protocol, please refer to Diana Cadena Castaneda et al. (https://doi.org/10.1016/j.isci.2023.107374).

Spatiotemporally organized immunomodulatory response to SARS-CoV-2 virus in primary human broncho-alveolar epithelia (preprint)

The COVID-19 pandemic continues to be a health crisis with major unmet medical needs. The early responses from airway epithelial cells, the first target of the virus regulating the progression towards severe disease, are not fully understood. Primary human air-liquid interface cultures representing the broncho-alveolar epithelia were used to study the kinetics and dynamics of SARS-CoV-2 variants infection. The infection measured by nucleoprotein expression, was a late event appearing between day 4-6 post infection for Wuhan-like virus. Other variants demonstrated increasingly accelerated timelines of infection. All variants triggered similar transcriptional signatures, an early inflammatory/immune signature preceding a late type I/III IFN, but differences in the quality and kinetics were found, consistent with the timing of nucleoprotein expression. Response to virus was spatially organized: CSF3 expression in basal cells and CCL20 in apical cells. Thus, SARS-CoV-2 virus triggers specific responses modulated over time to engage different arms of immune response.

Clinical Implications of Mask Wearing for Special Populations in an Inpatient Rehabilitation Setting during COVID-19 Global Pandemic

Research Objectives To describe the clinical implications of wearing a mask among special populations in inpatient rehabilitation. Design Narrative Review. Setting Inpatient rehabilitation. Participants Patients undergoing acute inpatient rehabilitation. Interventions Mask wearing. Main Outcome Measures Effect of mask wearing on exercise outcomes, including perceived exertion, shortness of breath, and patient perception;clinical implications for the interdisciplinary team. Results Five trials which observed exercise outcomes after mask wearing were identified and summarized. Findings for the effect of face masks on exercise and vital signs varied due to heterogeneity of exercise protocols and outcome measures. Relevant findings show that patients who wore masks had higher ratings of perceived exertion, increased shortness of breath, and feelings of claustrophobia while wearing a face mask during exercise. Wearing masks affected physical and mental outcomes despite exercise intensity (low, moderate, or high). Conclusions Moderate-to-high intensity exercise is needed for neuroplasticity. This type of exercise may not be possible in neurological populations, who do not have normal oxygen consumption at rest or with exercise, and may be further affected by wearing masks. Clinicians should their patients' mask wearing into consideration, and change the frequency, intensity, timing and types of exercises when patients are wearing a face mask. Frequent measurement of vital signs and fatigue symptoms are critical to maintain patient safety, and adjustments should be made as necessary. Other neurological challenges to monitor include impaired cognitive function, limited communication, reduced mood, and interactions with medications. These can affect the patients' ability to accurately determine and state their perceived effort or may hinder physiological and autonomic responses. Lastly, interdisciplinary teams should communicate patient findings related to mask wearing at team conferences, huddles and include discussion for ways to increase patients' participation and how to optimize health outcomes. Author(s) Disclosures None.