Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients.

Background: The global outbreak of COVID-19, and the limited availability of clinical treatments, forced researchers around the world to search for the pathogenesis and potential treatments. Understanding the pathogenesis of SARS-CoV-2 is crucial to respond better to the current coronavirus disease 2019 (COVID-19) pandemic. Methods: We collected sputum samples from 20 COVID-19 patients and healthy controls. Transmission electron microscopy was used to observe the morphology of SARS-CoV-2. Extracellular vesicles (EVs) were isolated from sputum and the supernatant of VeroE6 cells, and were characterized by transmission electron microscopy, nanoparticle tracking analysis and Western-Blotting. Furthermore, a proximity barcoding assay was used to investigate immune-related proteins in single EV, and the relationship between EVs and SARS-CoV-2. Result: Transmission electron microscopy images of SARS-COV-2 virus reveal EV-like vesicles around the virion, and western blot analysis of EVs extracted from the supernatant of SARS-COV-2-infected VeroE6 cells showed that they expressed SARS-COV-2 protein. These EVs have the infectivity of SARS-COV-2, and the addition can cause the infection and damage of normal VeroE6 cells. In addition, EVs derived from the sputum of patients infected with SARS-COV-2 expressed high levels of IL6 and TGF-ß, which correlated strongly with expression of the SARS-CoV-2 N protein. Among 40 EV subpopulations identified, 18 differed significantly between patients and controls. The EV subpopulation regulated by CD81 was the most likely to correlate with changes in the pulmonary microenvironment after SARS-CoV-2 infection. Single extracellular vesicles in the sputum of COVID-19 patients harbor infection-mediated alterations in host and virus-derived proteins. Conclusions: These results demonstrate that EVs derived from the sputum of patients participate in virus infection and immune responses. This study provides evidence of an association between EVs and SARS-CoV-2, providing insight into the possible pathogenesis of SARS-CoV-2 infection and the possibility of developing nanoparticle-based antiviral drugs.

Three-dimensional heterogeneous electro-Fenton system with reduced graphene oxide based particle electrode for Acyclovir removal

New pollutant pharmaceutical and personal care products (PPCPs), especially antiviral drugs, have received increasing attention not only due to their increase in usage after the outbreak of COVID-19 epidemics but also due to their adverse impacts on water ecological environment. Electro-Fenton technology is an effective method to remove PPCPs from water. Novel particle electrodes (MMT/rGO/Fe3O4) were synthesized by depositing Fe3O4 nanoparticles on reduced graphene oxide modified montmorillonite and acted as catalysts to promote oxidation performance in a three-dimensional Electro-Fenton (3D-EF) system. The electrodes combined the catalytic property of Fe3O4, hydrophilicity of montmorillonite and electrical conductivity of graphene oxides, and applied for the degradation of Acyclovir (ACV) with high efficiency and ease of operation. At optimal condition, the degradation rate of ACV reached 100% within 120 min, and the applicable pH range could be 3 to 11 in the 3D-EF system. The stability and reusability of MMT/rGO/Fe3O4 particle electrodes were also studied, the removal rate of ACV remained at 92% after 10 cycles, which was just slightly lower than that of the first cycle. Potential degradation mechanisms were also proposed by methanol quenching tests and FT-ICR-MS. Graphical Image, graphical Novel particle electrodes (MMT/rGO/Fe3O4) were synthesized by depositing Fe3O4 nanoparticles on reduced graphene oxide modified montmorillonite and acted as catalysts to promote oxidation performance in a three-dimensional electro-Fenton (3D-EF) system. Potential degradation mechanism and intermediate products were proposed.

U.S. Hospitals' Administrative Expenses Increased Sharply During COVID-19.

BACKGROUND: In response to the declining utilization and patient revenue due to the COVID-19 pandemic, the U.S. hospital industry furloughed at least 1.4 million health care workers to contain their clinical-related expenses. However, it remains unclear how hospitals responded by adjusting their administrative expenses, which account for more than a quarter of U.S. hospitals' spending, a proportion substantially higher than that of other industrialized countries. Examining changes in hospitals' administrative expenses during the COVID-19 pandemic is important for understanding hospitals' cost-containment behaviors under operational shocks during a pandemic. OBJECTIVE: To assess changes in hospitals' administrative expenses and clinical expenses during the COVID-19 pandemic in 2020. DESIGN: Time-series observational study. PARTICIPANTS: 1420 Medicare-certified general acute-care hospitals with fiscal years starting in January and continuously operating during 2016-2020. MAIN MEASURES: Hospitals' annual administrative expenses and clinical expenses. KEY RESULTS: Hospitals' median administrative and clinical expenses both increased consistently around 4% each year from 2016 to 2019. From 2019 to 2020, the median administrative expenses grew by 6.2% while the median clinical expenses grew by 0.6%. The interrupted time-series regression estimated an additional 6.4% (95% CI, 4.5 to 8.2%) increase in administrative expenses in 2020, relative to the pre-COVID annual increase of 3.9% (95% CI, 3.3 to 4.4%), while an additional increase in clinical expenses in 2020 (0.5%; 95% CI, -0.3 to 1.4%) did not differ from the pre-COVID annual increase of 3.7% (95% CI, 3.5 to 4%). Stratified analysis showed hospitals with larger utilization volume, located in states with lower COVID-19 burden, or situated in counties with higher median household income experienced larger increase in administrative expenses in 2020. CONCLUSIONS: In 2020, administrative expenses grew much faster than clinical expenses, resulting in a larger share of hospital financial resources allocated to administrative activities. Higher administrative expenses might reflect hospitals' operational effort in response to the pandemic or inefficient cost management.

COVID-19 and Hospital Financial Viability in the US.

Importance: The COVID-19 pandemic has had a negative association with hospital operations. To help health care facilities and clinicians stay financially viable during the COVID-19 pandemic, Congress provided $175 billion in subsidies. It remains unclear how much financial losses hospitals incurred owing to operational disruptions during the COVID-19 pandemic and whether subsidies were sufficient to offset the financial losses. Objective: To assess changes in the operational financial performance and overall financial viability of hospitals during the COVID-19 pandemic. Design Setting and Participants: This cross-sectional study included 1378 US hospitals whose fiscal years began in January and 785 hospitals whose fiscal years began in July (all with continuous observations from 2016 through 2020). RAND Hospital Data, a compiled and processed version of Medicare Cost Reports, were used. The data were analyzed on March 12, 2022. Exposures: The operational disruptions experienced and relief funds received by US hospitals during the COVID-19 pandemic. Main Outcomes and Measures: A hospital's annual operating margin, overall profit margin, and other nonoperating income as a share of total revenue from January 2016 to December 2020. Results: Among the 1378 hospitals with fiscal years beginning in January, the mean operating margin declined from -1.0% (95% CI,-1.9% to -0.1%) in 2019 to -7.4% (95% CI, -8.5% to -6.3%) in 2020. The mean share of other nonoperating income grew from 4.4% (95% CI, 4% to 4.7%) in 2019 to 10.3% (95% CI, 9.9% to 10.8%) in 2020. The mean overall profit in 2020 (6.7%; 95% CI, 5.4% to 8.1%) remained as stable as prior years. Government, rural, and smaller hospitals showed higher mean overall profit margins in 2020 than in 2019 (7.2% vs 3.7%, 7.5% vs 1.9%, and 6.7% vs 3.5%, respectively). These results remained consistent when hospitals whose fiscal years began in July were examined. Conclusions and Relevance: The results of this cross-sectional study suggest that although hospitals experienced a sizeable reduction in operating margins in 2020, their overall profit margins remained similar to those in prior years, suggesting that the COVID-19 relief fund effectively offset the financial losses for hospitals during the COVID-19 pandemic. Government, rural, and smaller hospitals, which were supported by some targeted fund allocations, generated higher overall profit margins during 2020 than in prior years.

Comparison of resident COVID-19 mortality between unionized and nonunionized private nursing homes.

Using bargaining agreement data from the Federal Mediation Conciliation Services, we found that the median national resident COVID-19 mortality percentage (as of April 24, 2022) of unionized nursing homes and that of nonunionized ones were not statically different (10.2% vs. 10.7%; P = 0.32). The median nursing home resident COVID-19 mortality percentage varied from 0% in Hawaii to above 16% in Rhode Island (16.6%). Unionized nursing homes had a statistically significant lower median mortality percentage than nonunionized nursing homes (P < 0.1) in Missouri, and had a higher median mortality percentage than nonunionized nursing homes (P < 0.05) in Alabama and Tennessee. Higher average resident age, lower percentage of Medicare residents, small size, for-profit ownership, and chain organization affiliation were associated with higher resident COVID-19 mortality percentage. Overall, no evidence was found that nursing home resident COVID-19 mortality percentage differed between unionized nursing homes and nonunionized nursing homes in the U.S.

Comparison of US Hospital Charity Care Policies Before vs After Onset of the COVID-19 Pandemic.

Importance: Tax-exempt hospitals are required to provide charity care to maintain their tax-exempt status; charity care policies must be published online with clear eligibility criteria. Prior research has shown wide variability in charity care policy content; it is unknown how hospitals change their charity care policies over time. Objective: To examine changes to tax-exempt hospital charity care policies before vs after the COVID-19 pandemic. Design, Setting, and Participants: This cohort study used downloaded charity care policies from a geographically representative sample of 170 tax-exempt hospitals from December 1 to 31, 2019, and December 1 to 31, 2021, and categorized the policy changes made as more restrictive, more generous, indeterminate, minimal, or not updated. Exposures: Onset of the COVID-19 pandemic. Main Outcomes and Measures: The primary outcome was charity care policy content changes from 2019 to 2021. Also examined were the effects of hospital ownership type, state Medicaid expansion status, and hospital consolidation on policy changes. Results: In this sample of 170 hospitals, 151 published documents available for comparison. Among these hospitals, 127 (84.1%) updated their charity care policies and 77 (51.0%) made substantial changes, with 242 distinct policy changes to categories such as income eligibility cutoffs, asset limitations, and service exclusions. Although the majority of hospitals expanded charity care (47 [31.1%]), a sizable minority restricted charity care (12 [7.9%]). Medicaid expansion during the study period and hospital consolidations were not associated with expansion of charity care; the largest merger in this sample led to reduced charity care at all 4 hospitals involved. Conclusions and Relevance: Tax-exempt hospitals appear to have updated their policies with mostly positive changes during and after the onset of the COVID-19 pandemic; however, some hospitals restricted charity care in 2021 documents. Unpublicized or vague eligibility criteria may limit patients' understanding of charity care policies and conceal the full extent of charity care policy changes over time. Policy makers should consider requiring greater transparency and simplification for hospital charity care policies to ensure adequate access to care for uninsured and underinsured patients.

NIH funding of COVID-19 research in 2020: a cross-sectional study.

OBJECTIVE: This study aims to characterise and evaluate the National Institutes of Health's (NIH's) grant allocation speed and pattern of COVID-19 research. DESIGN: Cross-sectional study. SETTING: COVID-19 NIH RePORTER Dataset was used to identify COVID-19 relevant grants. PARTICIPANTS: 1108 grants allocated to COVID-19 research. MAIN OUTCOMES AND MEASURES: The primary outcome was to determine the number of grants and funding amount the NIH allocated for COVID-19 by research type and clinical/scientific area. The secondary outcome was to calculate the time from the funding opportunity announcement to the award notice date. RESULTS: The NIH awarded a total of 56 169 grants in 2020, of which 2.0% (n=1108) wwas allocated for COVID-19 research. The NIH had a US$45.3 billion budget that year, of which 4.9% (US$2.2 billion) was allocated to COVID-19 research. The most common clinical/scientific areas were social determinants of health (n=278, 8.5% of COVID-19 funding), immunology (n=211, 25.8%) and pharmaceutical interventions research (n=208, 47.6%). There were 104 grants studying COVID-19 non-pharmaceutical interventions, of which 2 grants studied the efficacy of face masks and 6 studied the efficacy of social distancing. Of the 83 COVID-19 funded grants on transmission, 5 were awarded to study airborne transmission of COVID-19 and 2 grants on transmission of COVID-19 in schools. The average time from the funding opportunity announcement to the award notice date was 151 days (SD: ±57.9). CONCLUSION: In the first year of the pandemic, the NIH diverted a small fraction of its budget to COVID-19 research. Future health emergencies will require research funding to pivot in a timely fashion and funding levels to be proportional to the anticipated burden of disease in the population.

Initial Public-Health Emergency Response to SARS and COVID-19 Pandemics in Mainland China: A Retrospective Comparative Study.

BACKGROUND: Severe acute respiratory syndrome (SARS) was reported first in China in 2003. The world is currently coping with coronavirus disease-2019 (COVID-19). We conducted a retrospective study to compare the initial public-health emergency response (PHER) to SARS and COVID-19 in mainland China. METHODS: A qualitative comparative study was conducted to compare the PHER timelines to SARS and COVID-19 by selecting six crucial time points. Besides, we explored the speed of spread, peak time and plateau period of SARS and COVID-19, respectively, by comparing the confirmed cases in the same time interval. RESULTS: The government of the People's Republic of China (PRC) accomplished the entire initial PHER to SARS in 127 days and for COVID-19 in 44 days. The speed of PHER for COVID-19 was 83 days faster. The peak time of SARS arose ~80 days later than that of COVID-19. Though the peak number of confirmed daily cases for COVID-19 was fivefold more than that of SARS, the onset of the stabilization period for COVID-19 was >2 months earlier than that of SARS. CONCLUSION: Overall, the speed of the initial PHER to COVID-19 pandemic was faster than that for SARS. Compared with the speed of hospital reporting and government policymaking, the speed of pathogen identification improved the most. The COVID-19 pandemic curve entered a plateau period earlier than the SARS pandemic curve, which suggests that the pandemic was controlled more effectively because of a timely PHER. The PRC government should emphasize improving the ability of hospitals to restrain infectious diseases by enhancing the direct reporting system and cultivating crisis management to empower relevant individuals to make timely scientific decisions.

Early Surveillance and Public Health Emergency Responses Between Novel Coronavirus Disease 2019 and Avian Influenza in China: A Case-Comparison Study.

Background: Since the novel coronavirus disease (COVID-19) has been a worldwide pandemic, the early surveillance and public health emergency disposal are considered crucial to curb this emerging infectious disease. However, studies of COVID-19 on this topic in China are relatively few. Methods: A case-comparison study was conducted using a set of six key time nodes to form a reference framework for evaluating early surveillance and public health emergency disposal between H7N9 avian influenza (2013) in Shanghai and COVID-19 in Wuhan, China. Findings: A report to the local Center for Disease Control and Prevention, China, for the first hospitalized patient was sent after 6 and 20 days for H7N9 avian influenza and COVID-19, respectively. In contrast, the pathogen was identified faster in the case of COVID-19 than in the case of H7N9 avian influenza (12 vs. 31 days). The government response to COVID-19 was 10 days later than that to avian influenza. The entire process of early surveillance and public health emergency disposal lasted 5 days longer in COVID-19 than in H7N9 avian influenza (46 vs. 41 days). Conclusions: The identification of the unknown pathogen improved in China between the outbreaks of avian influenza and COVID-19. The longer emergency disposal period in the case of COVID-19 could be attributed to the government's slower response to the epidemic. Improving public health emergency management could lessen the adverse social effects of emerging infectious diseases and public health crisis in the future.

A Non-Profit Approach to Address Foreign Dependence of Generic Drugs.

The COVID-19 pandemic has revealed the vulnerability of the US generic drug supply chain to foreign production. Many policies have been proposed to mitigate this vulnerability. In this article, we argue that nonprofit drug manufacturers have the potential to make important contributions.

Characteristics of and Public Health Emergency Responses to COVID-19 and H1N1 Outbreaks: A Case-Comparison Study.

BACKGROUND: Recently, the novel coronavirus disease (COVID-19) has already spread rapidly as a global pandemic, just like the H1N1 swine influenza in 2009. Evidences have indicated that the efficiency of emergency response was considered crucial to curb the spread of the emerging infectious disease. However, studies of COVID-19 on this topic are relatively few. METHODS: A qualitative comparative study was conducted to compare the timeline of emergency responses to H1N1 (2009) and COVID-19, by using a set of six key time nodes selected from international literature. Besides, we also explored the spread speed and peak time of COVID-19 and H1N1 swine influenza by comparing the confirmed cases in the same time interval. RESULTS: The government's entire emergency responses to the epidemic, H1N1 swine influenza (2009) completed in 28 days, and COVID-19 (2019) completed in 46 days. Emergency responses speed for H1N1 was 18 days faster. As for the epidemic spread speed, the peak time of H1N1 came about 4 weeks later than that of COVID-19, and the H1N1 curve in America was flatter than COVID-19 in China within the first four months after the disease emerged. CONCLUSIONS: The speed of the emergency responses to H1N1 was faster than COVID-19, which might be an important influential factor for slowing down the arrival of the peak time at the beginning of the epidemic. Although COVID-19 in China is coming to an end, the government should improve the public health emergency system, in order to control the spread of the epidemic and lessen the adverse social effects in possible future outbreaks.