Objective Patients with COVID-19 are at risk for laryngeal injury and dysfunction secondary to respiratory failure, prolonged intubation, and other unique facets of this illness. Our goal is to report clinical features and treatment for patients presenting with voice, airway, and/or swallowing concerns postacute COVID-19. Study Design Case series. Setting Academic tertiary care center. Methods Patients presenting with laryngeal issues following recovery from COVID-19 were included after evaluation by our laryngology team. Data were collected via retrospective chart review from March 1, 2020, to April 1, 2021. This included details of the patient’s COVID-19 course, initial presentation to laryngology, and subsequent treatment. Results Twenty-four patients met inclusion criteria. Twenty (83%) patients were hospitalized, and 18 required endotracheal intubation for a median (range) duration of 14 days (6-31). Ten patients underwent tracheostomy. Patients were evaluated at a median 107 days (32-215) after their positive SARS-CoV-2 test result. The most common presenting concerns were dysphonia (n = 19, 79%), dyspnea (n = 17, 71%), and dysphagia (n = 6, 25%). Vocal fold motion impairment (50%), early glottic injury (39%), subglottic/tracheal stenosis (22%), and posterior glottic stenosis (17%) were identified in patients who required endotracheal intubation. Patients who did not need intubation were most frequently treated for muscle tension dysphonia (67%). Conclusion Patients may develop significant voice, airway, and/or swallowing issues postacute COVID-19. These complications are not limited to patients requiring intubation or tracheostomy. Multidisciplinary laryngology clinics will continue to play an integral role in diagnosing and treating patients with COVID-19–related laryngeal sequelae.

Nef is an HIV-encoded accessory protein that enhances pathogenicity by down-regulating major histocompatibility class I (MHC-I) expression to evade killing by cytotoxic T lymphocytes (CTLs). A potent Nef inhibitor that restores MHC-I is needed to promote immune-mediated clearance of HIV-infected cells. We discovered that the plecomacrolide family of natural products restored MHC-I to the surface of Nef-expressing primary cells with variable potency. Concanamycin A (CMA) counteracted Nef at subnanomolar concentrations that did not interfere with lysosomal acidification or degradation and were nontoxic in primary cell cultures. CMA specifically reversed Nef-mediated down-regulation of MHC-I, but not CD4, and cells treated with CMA showed reduced formation of the Nef:MHC-I:AP-1 complex required for MHC-I down-regulation. CMA restored expression of diverse allotypes of MHC-I in Nef-expressing cells and inhibited Nef alleles from divergent clades of HIV and simian immunodeficiency virus, including from primary patient isolates. Lastly, we found that restoration of MHC-I in HIV-infected cells was accompanied by enhanced CTL-mediated clearance of infected cells comparable to genetic deletion of Nef. Thus, we propose CMA as a lead compound for therapeutic inhibition of Nef to enhance immune-mediated clearance of HIV-infected cells.

Highlights d Hematopoietic stem and progenitor cells can serve as longterm reservoirs of HIV d HSPCs harbor both infectious and defective proviral genomes d HSPCs are an important source of residual plasma virus in treated people d Clonally amplified HIV proviruses contribute to residual plasma virus

Objectives In-office serial intralesional steroid injections (SILSIs) have become a commonly used treatment for subglottic stenosis. We characterized the impact of SILSIs on the time between operating room visits and incidence of glucocorticoid systemic side effects. Study Design Retrospective case series. Setting Academic tertiary care center. Methods All patients with subglottic stenosis receiving SILSIs at 1 institution from 2016 to 2020 were included. Surgery-free interval was compared using paired t tests. Side effect incidence was calculated with Kaplan-Meier methodology for visualization. Results Nineteen patients and 207 procedures were included. The majority of patients were White (95%) and female (95%) and had idiopathic subglottic stenosis (53%). Mean surgery-free interval for all patients was 8.7 months (95% CI, 5.6-11.8) before initiating SILSIs. Of 11 patients with calculable surgery-free interval, 10 experienced improvement, with a mean surgery-free interval increase of 4.6 months (95% CI, 2.4-6.7). Seven patients have not required surgery since initiation of SILSIs, with a mean follow-up time of 28 months (95% CI, 25-31). Noncutaneous systemic side effects occurred at a mean 3.2 months (95% CI, 2.4-4.0) from first injection and included Cushing’s syndrome, increased intraocular pressure, central serous chorioretinopathy, and new insulin requirement in the setting of diabetes. Conclusions Ninety-one percent of patients who initiated SILSIs and had a subsequent return to the operating room experienced a mean 4.6-month increase in surgery-free interval. Systemic side effects of glucocorticoids occurred in 32% of patients after initiating SILSIs. This should be considered in preprocedure counseling and side effect monitoring during treatment.

Objectives To characterize the impact of subglottic stenosis (SGS) on voice‐related quality of life (V‐RQOL) and quantify the effect of treatment on voice outcomes. Study Design Case series. Methods Retrospective review of SGS patients treated from 1996 to 2018 at a single institution to assess for 1) V‐RQOL association with individual patient cumulative treatment number and 2) V‐RQOL correlation with treatment type, time between treatments, and degree of stenosis. Analysis included both parametric and nonparametric statistical comparison across treatment types and multivariable and univariate linear regression. Results Sixty‐one patients, predominantly white (93%) and female (93%), were included. Etiology of SGS included idiopathic (61%), iatrogenic (16%), granulomatosis with polyangiitis (16%), and other (7%). The plurality of patients had four or more treatments (44%), with the remainder having one (28%), two (13%), or three treatments (15%). Analysis of change between pre‐ and postoperative V‐RQOL scores was completed for 130 treatments. These included dilation with laser incision (52%), in‐office injection (34%), dilation without division (8%), cricotracheal resection (1%), and all other treatment (8%). For every 10% improvement in airway caliber postoperatively, there was a 1.3‐point improvement in calculated V‐RQOL (r = 0.27, P = .02). After adjustment for treatment type, age, sex, and SGS etiology, this association held (beta = 1.5, P = .02). Change in V‐RQOL was not associated with treatment type, treatment number, or time between treatments. Conclusion Patients with subglottic stenosis who have greater degree of change in airway caliber experience greater improvement in V‐RQOL scores following treatment. V‐RQOL scores are not associated with treatment type or time between individual treatments. Level of Evidence 4 Laryngoscope, 131:360–365, 2021

BackgroundThe COVID-19 pandemic limited in-person medical experiences for senior medical students. To address the lack of visiting sub-internships, we created a one-day virtual overview of Otolaryngology-Head and Neck Surgery, incorporating discussion of important trainee topics and an introduction to our residency program.MethodsA one-day virtual event was hosted by a tertiary academic medical center. Sessions were structured as interviews with faculty and residents, with real-time question and answer sessions. A guide with presenter information and resident-curated resources for each sub-specialty was given, as well as a document of frequently asked questions. The event was advertised to senior medical students through online forums. Data were collected via registration and post-event anonymous surveys. ResultsThere were 327 participants representing 138 institutions, 34 states and territories, and 10 countries, with 90% of participants consisting of senior medical students. The event spanned 5 content hours, and participants were signed in for 4.1 hours on average. Participants (n = 102, 31% response rate) found the event helpful for learning about the field (96%), residency program (97%), and specialty-specific resources (86%). Hearing from residents (43%) and faculty (41%) were cited as the most helpful parts of the program. Getting to know residency programs remotely (28%), participating in virtual interviews (26%), and not having away rotations (20%) were most often noted as the biggest student concerns for the 2020–2021 application cycle. Finally, participants identified getting to know an institution’s culture (76%) as the most difficult aspect of a program to evaluate remotely.ConclusionsWe demonstrate that a virtual event can be valuable in the transition to residency by supplementing or partially substituting visiting sub-internships.

A bacteriophage is a virus that infects and replicates in bacteria. To reproduce, the phage must convert the host cell metabolism from that suited for cell survival to one favorable for producing new phage progeny. One mechanism by which phage can cause host cell conversion is by expressing ‘cytotoxic’ phage gene products that interact with and inhibit the function of specific protein targets of key metabolic pathways of the bacterial host. Identifying ‘cytotoxic’ phage genes and understanding more about how phage exploit their host's weaknesses could lead to new and important therapies for many bacterial illnesses. In this work, a cytotoxic gene from mycobacteriophage Vix, gene 80, was studied. The protein of the Vix80 gene is 68% identical to the product of gene 77 of mycobacteriophage L5, a gene that has been shown to be cytotoxic to Mycobacterium. We confirmed that the Vix80 gene product is also cytotoxic when expressed in Mycobacterium smegmatis. Our hypothesis is that a physical interaction between the Vix80 gene product and a specific host cell protein affects cell metabolism and causes growth inhibition. A 6xHis tagged Vix80 protein was created and expressed in E. coli, and then purified by column chromatography. SDS‐PAGE and staining verified that we had expressed and purified the Vix80 protein of correct size. We are using the His‐tagged Vix80 protein in a biochemical approach to identify the interacting host cell protein. Supported by an ASBMB Undergraduate Research Award, and a Summer Research Scholar Award from Hope College Biology Department

Introduction: Abdominal Aortic Aneurysm (AAA) development is linked to smoking and is characterized by extracellular matrix remodeling by MMP2 and -9. We hypothesized that MMP2 and -9 secretions and regulation in response to cigarette smoke is modulated by interactions between resident cells within the aorta, namely, aortic smooth muscles, endothelial cells, and infiltrating macrophages and could influence AAA formation and/or progression. Methods: Aqueous extract of cigarette smoke (AEC) was applied to male rat aortic smooth muscle (RASMC), endothelial (RAEC) or RAW cells in serum free medium (SFM). After 24h the cells were washed, fresh SFM was added and collected after 24h. This conditioned media (CM) was applied to the cells for 24h, again washed, fresh SFM added, and then incubated for 72h, at which point the media was saved for MMP2 and -9 analysis by zymography while the cellular protein was used to determine the ratio (pS/pJ) of phospho-Stat3 (pStat3) and phospho-Jak2 (pJak2) by western blot. Results: Unstimulated CM from RAW cells increased MMP2 by 24% (p=0.0357), MMP9 by 282.1% (p=0.0004) and pS/pJ by 136.5% in RASMC. AEC-CM from RAW and RAEC cells increased MMP9 by 200.5% (p=0.0004) and 17.1% (p=0.0301) respectively in RASMC and corresponding increases in pS/pJ (305.7% and 227.6%, respectively). AEC-CM derived from RAW cells induced RAEC to produce moderate amounts of MMP2 (16.5%, p=0.0140) and MMP9 (29.5%, p=0.0207) and a 137.0% increase in pS/pJ. RAW cells receiving unstimulated CM from RASMC and RAEC cells produced significant amounts of MMP9 (127.7%, p=0.0026 and 236.8%, p=0.0020, respectively) and increased levels of pS/pJ ratios (45.2% and 1282.8%, respectively). Moreover, AEC-CM from RASMC and RAEC induced significant production of MMP9 from RAW cells (154.8%, p=0.0001 and 162.4%, p=0.0022, respectively) and increase in pS/pJ ratios (1348.2%, and 1493.9%, respectively) over baseline. Conclusion: This is the first in vitro study demonstrating that cigarette smoke results in differential interactions between aortic smooth muscle cells, endothelial cells, and macrophage lineage cells resulting in enhanced levels of MMP2, MMP9, pStat3 and pJak2. These interactions may play an important role in AAA formation under in vivo conditions.