The development of small-molecules targeting different components of SARS-CoV-2 is a key strategy to complement antibody-based treatments and vaccination campaigns in managing the COVID-19 pandemic. ...Here, we show that two thiol-based chemical probes that act as reducing agents, P2119 and P2165, inhibit infection by human coronaviruses, including SARS-CoV-2, and decrease the binding of spike glycoprotein to its receptor, the angiotensin-converting enzyme 2 (ACE2). Proteomics and reactive cysteine profiling link the antiviral activity to the reduction of key disulfides, specifically by disruption of the Cys379-Cys432 and Cys391-Cys525 pairs distal to the receptor binding motif in the receptor binding domain (RBD) of the spike glycoprotein. Computational analyses provide insight into conformation changes that occur when these disulfides break or form, consistent with an allosteric role, and indicate that P2119/P2165 target a conserved hydrophobic binding pocket in the RBD with the benzyl thiol-reducing moiety pointed directly toward Cys432. These collective findings establish the vulnerability of human coronaviruses to thiol-based chemical probes and lay the groundwork for developing compounds of this class, as a strategy to inhibit the SARS-CoV-2 infection by shifting the spike glycoprotein redox scaffold.
Music-based interventions (MBIs) show promise for managing symptoms of various brain disorders. To fully realize the potential of MBIs and dispel the outdated misconception that MBIs are rooted in ...soft science, the NIH is promoting rigorously designed, well-powered MBI clinical trials. The pressing need of guidelines for scientifically rigorous studies with enhanced data collection brought together the Renée Fleming Foundation, the Foundation for the NIH, the Trans-NIH Music and Health Working Group, and an interdisciplinary scientific expert panel to create the NIH MBI Toolkit for research on music and health across the lifespan. The Toolkit defines the building blocks of MBIs, including a consolidated set of common data elements for MBI protocols, and core datasets of outcome measures and biomarkers for brain disorders of aging that researchers may select for their studies. Utilization of the guiding principles in this Toolkit will be strongly recommended for NIH-funded studies of MBIs.
The incidence and sites of mucus accumulation and molecular regulation of mucin gene expression in coronavirus (COVID-19) lung disease have not been reported.
To characterize the incidence of mucus ...accumulation and the mechanisms mediating mucin hypersecretion in COVID-19 lung disease.
Airway mucus and mucins were evaluated in COVID-19 autopsy lungs by Alcian blue and periodic acid-Schiff staining, immunohistochemical staining, RNA
hybridization, and spatial transcriptional profiling. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected human bronchial epithelial (HBE) cultures were used to investigate mechanisms of SARS-CoV-2-induced mucin expression and synthesis and test candidate countermeasures.
MUC5B and variably MUC5AC RNA concentrations were increased throughout all airway regions of COVID-19 autopsy lungs, notably in the subacute/chronic disease phase after SARS-CoV-2 clearance. In the distal lung, MUC5B-dominated mucus plugging was observed in 90% of subjects with COVID-19 in both morphologically identified bronchioles and microcysts, and MUC5B accumulated in damaged alveolar spaces. SARS-CoV-2-infected HBE cultures exhibited peak titers 3 days after inoculation, whereas induction of MUC5B/MUC5AC peaked 7-14 days after inoculation. SARS-CoV-2 infection of HBE cultures induced expression of epidermal growth factor receptor (EGFR) ligands and inflammatory cytokines (e.g., IL-1α/β) associated with mucin gene regulation. Inhibiting EGFR/IL-1R pathways or administration of dexamethasone reduced SARS-CoV-2-induced mucin expression.
SARS-CoV-2 infection is associated with a high prevalence of distal airspace mucus accumulation and increased MUC5B expression in COVID-19 autopsy lungs. HBE culture studies identified roles for EGFR and IL-1R signaling in mucin gene regulation after SARS-CoV-2 infection. These data suggest that time-sensitive mucolytic agents, specific pathway inhibitors, or corticosteroid administration may be therapeutic for COVID-19 lung disease.
To effectively combat the simultaneous overdose and maternal mortality crises, a multimodal approach is needed. The aim of this study is to evaluate the preliminary effectiveness of a pilot, ...experiential learning, substance use disorder (SUD) curriculum embedded into a third-year medical student obstetrics and gynecology clerkship to improve self-reported confidence in SUD clinical skills.
This SUD curriculum was designed and implemented in an outpatient clinic, which provides integrated obstetric, gynecologic, and addiction medicine services for pregnant and parenting people with SUD. Third-year medical students on their obstetrics and gynecology clerkship rotated 1 full day through the OB MOTIVATE clinic between August 2020 and April 2022 and completed this curriculum. Students completed preclinic assignments and in-clinic tasks (eg, practicing SBIRT under supervision: screening, brief intervention, referral to treatment). Paired t tests assessed changes in outcomes, with increasing scores (range 1-5) demonstrating improvement.
Sixty-three students rotated through the OB MOTIVATE clinic; 57 completed the curriculum and surveys. Results from the self-assessment tools demonstrated significant improvements in confidence in SUD clinical skills, including performing SBIRT (2.46 ± 0.80 vs 4.07 ± 0.59, P < 0.01), motivational interviewing (2.98 ± 0.86 vs 4.16 ± 0.65, P < 0.01), using evidence-based medicine (2.91 ± 1.09 vs 4.23 ± 0.66, P < 0.01), and collecting an SUD history (3.25 ± 1.04 vs 4.35 ± 0.55, P = 0.01).
The integration of interventional curriculums into medical school and residency programs could be an effective avenue to reinforce addiction knowledge and teach new skills. This practical 1-day pilot curriculum demonstrated preliminary effectiveness at introducing third-year medical students to the complexities of SUD in pregnancy and postpartum. Further investigations of feasible and acceptable SUD educational interventions are warranted.
Abstract
Migration is increasingly common in Africa, especially for employment. Migrants may face additional barriers to accessing health care, including human immunodeficiency virus (HIV) prevention ...and treatment, compared with long-term residents. Exploring migrants’ experiences with health services can provide insights to inform the design of health programmes. In this study, we used qualitative methods to understand migrants’ barriers to health service utilization in south-central Uganda. This secondary data analysis used data from in-depth semi-structured interviews with 35 migrants and 25 key informants between 2017 and 2021. Interviews were analysed thematically through team debriefings and memos. We constructed three representative migrant journeys to illustrate barriers to accessing health services, reflecting experiences of migrant personas with differing HIV status and wealth. Migrants reported experiencing a range of barriers, which largely depended on the resources they could access, their existing health needs and their ability to form connections and relationships at their destination. Migrants were less familiar with local health services, and sometimes needed more time and resources to access care. Migrants living with HIV faced additional barriers to accessing health services due to anticipated discrimination from community members or health workers and difficulties in continuing antiretroviral therapy when switching health facilities. Despite these barriers, social networks and local connections facilitated access. However, for some migrants, such as those who were poorer or living with HIV, these barriers were more pronounced. Our work highlights how local connections with community members and health workers help migrants access health services. In practice, reducing barriers to health services is likely to benefit both migrants and long-term residents.
Oesophageal stents are meshed tubular implants designed to maintain patency of the oesophageal lumen and attenuate the symptoms of oesophageal cancer. Oesophageal cancers account for one in twenty ...cancer diagnoses and can lead to dysphasia, malnutrition and the diminishment of patient quality of life (QOL). Self-expanding oesophageal stents are the most common approach to attenuate these symptoms. Recent advances in oncological therapy have enabled patient survival beyond the lifetime of current devices. This introduces new complications for palliation, driving the need for innovation in stent design. This review identifies the factors responsible for stent failure. It explores the challenges of enhancing the longevity of stent therapies and outlines solutions to improving clinical outcomes. Discussions focus on the role of stent materials, construction methods, and coatings upon device performance. We found three key stent enhancement strategies currently used; material surface treatments, anti-migratory modifications, and biodegradable skeletons. Furthermore, radioactive and drug eluting stent designs were identified as emerging novel treatments. In conclusion, the review offers an overview of remaining key challenges in oesophageal stent design and potential solutions. It is clear that further research is needed to improve the clinical outcome of stents and patient QOL.
Human African trypanosomiasis (HAT) is a neglected tropical disease caused by the protozoan parasite Trypanosoma brucei. Because drugs in use against HAT are toxic and require intravenous dosing, new ...drugs are needed. Initiating lead discovery campaigns by using chemical scaffolds from drugs approved for other indications can speed up drug discovery for neglected diseases. We demonstrated recently that the 4-anilinoquinazolines lapatinib (GW572016, 1) and canertinib (CI-1033) kill T. brucei with low micromolar EC50 values. We now report promising activity of analogues of 1, which provided an excellent starting point for optimization of the chemotype. Our compound optimization that has led to synthesis of several potent 4-anilinoquinazolines, including NEU617, 23a, a highly potent, orally bioavailable inhibitor of trypanosome replication. At the cellular level, 23a blocks duplication of the kinetoplast and arrests cytokinesis, making it a new chemical tool for studying regulation of the trypanosome cell cycle.
Identification of fungal organisms often poses a problem for pathologists because the histomorphology of some fungal organisms is not specific, fresh tissues may not be available, and isolation and ...identification in culture may take a long time. The purpose of this study was to validate the use of panfungal polymerase chain reaction (PCR) to identify fungal organisms from formalin-fixed paraffin-embedded (FFPE) tissues. Formalin-fixed paraffin-embedded curls were tested from 128 blocks containing canine, feline, equine, and bovine tissues with cutaneous, nasal, pulmonary, and systemic fungal infections, identified by the presence of fungi in histologic sections. Quantitative scoring of histologic sections identified rare (11.9%), occasional (17.5%), moderate (17.5%), or abundant (53.1%) fungal organisms. DNA was isolated from FFPE tissues and PCR was performed targeting the internal transcribed spacer 2 (ITS-2) region, a segment of noncoding DNA found in all eukaryotes. Polymerase chain reaction products were sequenced and identified at ≥97% identity match using the Basic Local Alignment Search Tool and the NCBI database of ITS sequences. Of the 128 blocks, 117 (91.4%) yielded PCR products and high-quality sequences were derived from 89 (69.5%). Sequence and histologic identifications matched in 79 blocks (61.7%). This assay was capable of providing genus- and species-level identification when histopathology could not and, thus, is a beneficial complementary tool for diagnosis of fungal diseases.
The severe acute respiratory syndrome coronavirus 2 pandemic is characterized by the emergence of novel variants of concern (VOCs) that replace ancestral strains. Here, we dissect the complex ...selective pressures by evaluating variant fitness and adaptation in human respiratory tissues. We evaluate viral properties and host responses to reconstruct forces behind D614G through Omicron (BA.1) emergence. We observe differential replication in airway epithelia, differences in cellular tropism, and virus-induced cytotoxicity. D614G accumulates the most mutations after infection, supporting zoonosis and adaptation to the human airway. We perform head-to-head competitions and observe the highest fitness for Gamma and Delta. Under these conditions, RNA recombination favors variants encoding the B.1.617.1 lineage 3′ end. Based on viral growth kinetics, Alpha, Gamma, and Delta exhibit increased fitness compared to D614G. In contrast, the global success of Omicron likely derives from increased transmission and antigenic variation. Our data provide molecular evidence to support epidemiological observations of VOC emergence.
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•The ancestral D614G is least adapted to the human airway compared to later variants•Gamma and Delta variants have the highest replicative fitness in human airway epithelia•Omicron displays poor replication and increased immune activation in the lower airway•Omicron success globally is likely due to increased transmission and antigenic variation
Meganck et al. recapitulate the drivers of evolution by evaluating SARS-CoV-2 variant fitness in primary human airway cells. Ancestral D614G displays pre-adaptation phenotypes, while Gamma and Delta variants have intrinsic high replicative fitness in the human airway. Omicron has poor replicative fitness and greater immune induction in the lower airway.
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