BackgroundPre-specified interim analyses allow for more timely evaluation of efficacy or futility, potentially accelerating decision-making on an investigational intervention. In such an analysis, the randomized, double-blind MOVe-OUT trial demonstrated superiority of molnupiravir over placebo for outpatient treatment of COVID-19 in high-risk patients. In the full analysis population, the point estimate of the treatment difference in the primary endpoint was notably lower than at the interim analysis. We conducted a comprehensive assessment to investigate this unexpected difference in treatment effect size, with the goal of informing future clinical research evaluating treatments for rapidly evolving infectious diseases.MethodsThe modified intention-to-treat population of the MOVe-OUT trial was divided into an interim analysis cohort (i.e. all participants included in the interim analysis; prospectively defined) and a post-interim analysis cohort (i.e. all remaining participants; retrospectively defined). Baseline characteristics (including many well-established prognostic factors for disease progression), clinical outcomes, and virologic outcomes were retrospectively evaluated. The impact of changes in baseline characteristics over time was explored using logistic regression modeling and simulations.ResultsBaseline characteristics were well-balanced between arms overall. However, between- and within-arm differences in known prognostic baseline factors (e.g. comorbidities, SARS-CoV-2 viral load, and anti-SARS-CoV-2 antibody status) were observed for the interim and post-interim analysis cohorts. For the individual factors, these differences were generally minor and otherwise not notable; as the trial progressed, however, these shifts in combination increasingly favored the placebo arm across most of the evaluated factors in the post-interim cohort. Model-based simulations confirmed that the reduction in effect size could be accounted for by these longitudinal trends toward a lower-risk study population among placebo participants. Infectivity and viral load data confirmed that molnupiravir's antiviral activity was consistent across both cohorts, which were heavily dominated by different viral clades (reflecting the rapid SARS-CoV-2 evolution).DiscussionThe cumulative effect of randomly occurring minor differences in prognostic baseline characteristics within and between arms over time, rather than virologic factors such as reduced activity of molnupiravir against evolving variants, likely impacted the observed outcomes. Our results have broader implications for group sequential trials seeking to evaluate treatments for rapidly emerging pathogens. During dynamic epidemic or pandemic conditions, adaptive trials should be designed and interpreted especially carefully, considering that they will likely rapidly enroll a large post-interim overrun population and that even small longitudinal shifts across multiple baseline variables can disproportionately impact prespecified efficacy outcomes at different timepoints. Shifts in prognostic factors may introduce additional variability that can be difficult to disentangle from temporal trends in epidemiology (e.g. evolutionary changes in the causative pathogen) or disease management.(ClinicalTrials.gov: NCT04575597.).

The prolonged viral shedding from the gastrointestinal tract is well documented for numerous pathogens, including SARS-CoV-2. However, the impact of prolonged viral shedding on epidemiological inferences using wastewater data is not yet fully understood.

To gain a better understanding of this phenomenon at the population level, we extended a wastewater-based modeling framework that integrates viral shedding dynamics, viral load data in wastewater, case report data, and an epidemic model.

Our results indicate that as an outbreak progresses, the viral load from recovered individuals gradually becomes predominant, surpassing that from the infectious population. This phenomenon leads to a dynamic relationship between model-inferred and reported daily incidence over the course of an outbreak. Sensitivity analyses on the duration and rate of viral shedding for recovered individuals reveal that accounting for this phenomenon can considerably advance prediction of transmission peak timing. Furthermore, extensive viral shedding from the recovered population toward the conclusion of an epidemic wave may overshadow viral signals from newly infected cases carrying emerging variants, which can delay the rapid recognition of emerging variants based on viral load.

These findings highlight the necessity of integrating post-recovery viral shedding to enhance the accuracy and utility of wastewater-based epidemiological analysis.

Coronavirus disease 2019 (COVID-19) is often associated with thrombosis. Elevated levels of soluble C-type lectin-like receptor 2 (sCLEC-2), a biomarker for platelet activation, have been reported in COVID-19. Therefore, we examined the behavior of sCLEC-2 levels and their relationship with thrombosis.The clinical course of inflammatory and thrombotic biomarkers was assessed in 271 patients with COVID-19.Inflammatory biomarkers such as C-reactive protein, procalcitonin, and presepsin levels were significantly increased in patients with COVID-19, and these behaviors differed among the clinical course or stages. The plasma D-dimer levels increased slightly and gradually. Platelet counts were within the normal range, and plasma sCLEC-2 levels were markedly increased in most patients with COVID-19. There were 17 patients with thrombosis in this study. Although there was no significant difference in various biomarkers between COVID-19 patients with and without thrombosis, the super formula of sCLEC-2xD-dimer/platelet count in patients with thrombosis was significantly higher than in those without thrombosis. Furthermore, this super formula was significantly higher in COVID-19 patients with severe or critical illness than in those with mild or moderate illness.Elevation of the super formula of sCLEC-2xD-dimer/platelet count was associated with the thrombosis in patients with COVID-19 suggesting the thrombosis in COVID-19 may be caused by the development of microthrombosis.

During the COVID-19 pandemic, several studies were published on the use of ozone therapy in treating COVID-19, leveraging pre-pandemic published data on the anti-inflammatory, antimicrobial, and immunomodulatory properties of ozone gas. In this pilot randomized controlled trial conducted during the pandemic, we aimed to assess the outcomes of blood ozone therapy (OT) as an investigational agent versus the COVID-19 standard of care as standalone on 60 patients diagnosed with severe COVID-19.

This study was conducted as a randomized controlled trial in which both arms of the study received the Iranian Health Ministry's COVID-19 treatment guideline as the standard of care; the intervention group additionally received intravenous ozonated blood based on the related international society guidelines.

Our findings revealed a statistically non-significant 33% higher hazard ratio for a prolonged hospital stay in the OT group. However, the OT arm exhibited a significantly higher odds ratio of 4.3 for ICU transfer of patients initially admitted to general wards. The univariate logistic regression analysis of mortality found a 3.5-fold increased probability associated with OT use, though this difference was not statistically significant.

We suggest that further trials with robust study designs utilizing larger populations are required to further assess the role of OT on severe COVID-19 keeping in mind a heightened awareness of potential unfavorable outcomes throughout the study.

https://irct.ir, identifier IRCT20200616047792N1.

SARS-CoV-2 infection frequently involves the respiratory system and may impact on pulmonary function tests (PFT) of recovered individuals. Studies which compare post-COVID-19 PFT to pre-illness measurements are scarce. The primary objective of this study was to assess the effect of COVID-19 on PFT soon after infection.

In this prospective observational study, PFT were measured early following recovery from COVID-19 among healthy military aircrew. Spirometry values were compared to pre-COVID-19 measurements, and abnormality rates of lung volumes and diffusion capacity for carbon monoxide (DLCO) were assessed.

The study included 252 aviators, 97.6% males, mean age 34.9-years, following recovery from SARS-CoV-2 infection. Participants manifested mild symptoms (79.4%) or were asymptomatic (20.6%). Post-COVID-19 spirometry results 10.79 ± 5.67 days following infection were compared to measurements performed 41.3 ± 28.59 months earlier. Pre- and post-COVID-19 results were comparable, with similar minimal abnormalities rates (2% and 4.4%, respectively). In addition, there were no restrictive abnormalities following infection, and just 7.7% of individuals had a marginally low DLCO of 70-80% of predicted.

Among vaccinated, healthy adults, mild COVID-19 had no significant impact on PFT early post-infection. The data suggest that systematic PFT testing might not be necessary for asymptomatic healthy individuals who recovered from mild COVID-19.

The objective was to investigate the implications of enteral nutrition for elderly patients with common-type coronavirus disease 2019 (COVID-19).

Data were retrospectively extracted from medical records. Enteral nutritional supplementation was recommended for patients with a nutritional risk score >3. The preferred method was oral administration, and preparations included Ensure and TPF-T. Continuous variables were compared using analysis of two-tailed Student's t-tests or one-way analysis of variance for normally distributed data and the rank sum test for non-normally distributed data. Categorical variables were compared using the χ2 test or Fisher's exact test. Values of p <0.05 were considered to be statistically significant.

The mortality rate in the whole cohort was 9.54%. A total of 474 patients tested negative and were discharged; among them, 173 patients received enteral nutrition while 301 patients did not. There were significant correlations between mortality and age, serum albumin concentration, prognostic nutritional index, underlying severe disease status and diet condition. In patients with a poor diet, early use of enteral nutrition is associated with faster conversion to a negative polymerase chain reaction test.

The prognosis of elderly patients with common-type COVID-19 was related to their nutritional status. Enteral nutritional supplementation is the preferred method of nutrition because it is the simplest and most widely accepted method for patients. For patients with poor diet conditions, enteral nutritional intervention should be performed early.

The COVID-19 pandemic presents significant future health challenges. Its impact on pregnant women and their newborn is a particular area of concern. This study aims to examine the potential role of maternal immune activation (MIA), due to SARS-CoV-2 infection, on early neurodevelopment.

We analysed 107 mother-infant dyads from the COGESTCOV-19 study in Cantabria, Spain, which included 59 SARS-CoV-2 exposed (cases) and 48 unexposed (controls) mothers, recruited between December 2020 and February 2022. Cytokine levels (IL-6 and IL-10) were obtained from maternal blood and cord blood. Neurodevelopment was assessed using the Neonatal Behavioral Assessment Scale (NBAS) at six weeks of age. Trimester of infection was considered in the main analyses.

Results showed no significant overall delays in early neurodevelopment due to maternal SARS-CoV-2 infection. Control infants performed better in some NBAS items. However, cases infants showed trimester-specific differences. First-trimester exposure was related to motor and reflex delays, second-trimester to poorer performances in motor tasks and autonomic stability, and third-trimester to weaker state organization, regulation, and reflexes. Some correlations between cytokine levels and NBAS performance showed moderate associations.

These findings highlight the need for ongoing neurodevelopmental monitoring of infants born during the COVID-19 pandemic. The study enhances our understanding of MIA's impact on early development, emphasizing the importance of addressing homeostatic mechanisms in mothers and newborns.

COVID-19 exhibits complex pathophysiological manifestations, characterized by significant clinical and biological heterogeneity. Identifying phenotypes may enhance our understanding of the disease's diverse trajectories, benefiting clinical practice and trials.

This study included adult patients with COVID-19 from Xinhua Hospital, affiliated with Shanghai Jiao Tong University School of Medicine, between December 15, 2022, and February 15, 2023. The k-prototypes clustering method was employed using 50 clinical variables to identify phenotypes. Machine learning algorithms were then applied to select key classifier variables for phenotype recognition.

A total of 1376 patients met the inclusion criteria. K-prototypes clustering revealed two distinct subphenotypes: Hypo-inflammatory subphenotype (824 [59.9%]) and Hyper-inflammatory subphenotype (552 [40.1%]). Patients in Hypo-inflammatory subphenotype were younger, predominantly female, with low mortality and shorter hospital stays. In contrast, Hyper-inflammatory subphenotype patients were older, predominantly male, exhibiting a hyperinflammatory state with higher mortality and rates of organ dysfunction. The AdaBoost model performed best for subphenotype prediction (Accuracy: 0.975, Precision: 0.968, Recall: 0.976, F1: 0.972, AUROC: 0.975). "CRP", "IL-2R", "D-dimer", "ST2", "BUN", "NT-proBNP", "neutrophil percentage", and "lymphocyte count" were identified as the top-ranked variables in the AdaBoost model.

This analysis identified two phenotypes based on COVID-19 symptoms and comorbidities. These phenotypes can be accurately recognized using machine learning models, with the AdaBoost model being optimal for predicting in-hospital mortality. The variables "CRP", "IL-2R", "D-dimer", "ST2", "BUN", "NT-proBNP", "neutrophil percentage", and "lymphocyte count" play a significant role in the prediction of subphenotypes. Use the identified subphenotypes for risk stratification in clinical practice. Hyper-inflammatory subphenotypes can be closely monitored, and preventive measures such as early admission to the intensive care unit or prophylactic anticoagulation can be taken.

Background and Objectives: The pulmonary sequelae of COVID-19 and their evolution are of interest to the scientific community. We aimed to determine the radiological changes at 6 and 12 months after COVID-19 pneumonia, its evolution and its risk factors. Materials and Methods: This retrospective longitudinal study included adults admitted for COVID-19 pneumonia from 1 March 2020 to 30 April 2021 who had a high-resolution computed tomography (HRCT) scan at 6 months and 12 months after hospital discharge. The primary outcome was the appearance of radiological abnormalities on HRCT and the number of lung segments affected by them at 6 and 12 months, while the main explanatory variables were about the disease course, analytical parameters and treatment. Results: This study included n = 108 patients, with a mean age of 64 years. There was a decrease in the percentage of patients presenting parenchymal (93.5% to 88.9%, p < 0.001) and reticular (63% to 62%, p < 0.001) patterns on HRCT at 12 months compared to 6, and an increase in those presenting a fibrotic pattern (62% to 63.9%, p < 0.001). Ground-glass opacities were the most frequent radiological change at 6 and 12 months (91.7% and 87%, respectively). There was a significant reduction in the total number of lung segments with ground-glass opacities (445 to 382, p < 0.001) and consolidation (158 to 136, p = 0.019) and an increase in those with bronchiectasis (66 to 80, p = 0.033) between the two moments. After multivariate analysis, high-flow oxygen therapy (HFOT), highest ferritin levels, hypertension and ≥71 years showed an association with the development of subpleural parenchymal bands, consolidation, bronchiectasis and septal thickening at 6 and 12 months. Conclusions: Parenchymal patterns seem to be more frequent than reticular and fibrotic patterns after COVID-19 pneumonia. The fibrotic pattern was the only one that worsened significantly over time, with bronchiectasis being the only change that increased at 12 months. Older age, hypertension, the need for HFOT, and high levels of ferritin may be directly associated with worse radiological outcomes after COVID-19 pneumonia.

Background: The US Food and Drug Administration (FDA) and International Organization for Standardization (ISO) clearance standards for the clinical use of smart device pulse oximetry require in-laboratory human hypoxemia testing in healthy human individuals using arterial blood gas analysis. Methods: We evaluated the SpO2 measurements of the Samsung smartphone (Galaxy S9/10) and smartwatch (Galaxy 4) at stable arterial oxygen saturations (SaO2) between 70 and 100% in 24 healthy participants. Testing followed FDA/ISO-stipulated procedures for pulse oximetry performance validation, which include questionnaire estimation of skin tone based on Fitzpatrick estimation of skin types I-VI. During testing, inspired oxygen, nitrogen, and carbon dioxide partial pressures were monitored and adjusted via partial rebreathing circuits to achieve stable target arterial blood oxygen (SaO2) plateaus between 70% and 100%. Arterial blood samples were taken at each plateau, with device SpO2 readings taken at each sample extraction. An ABL-90FLEX blood gas analyzer determined arterial blood sample SaO2. Bias, calculated from device readings minus corresponding arterial blood measurements, was reported as root mean square deviation (RMSD). Results: Combined Participants demographics were: 62.5% female; median age 26 years (range 21-46); and race/ethnicity 16.7% African American, 33.3% Asian, 12.5% multi-ethnic, and 37.5% Caucasian. Fitzpatrick Skin Scale-identified skin tones were: white-fair (I&II), 20.8%; average-light brown (III-IV), 54% and brown-black (V-VI), 25%. There were no adverse events. The RMSD values of SpO2 measurements were: smartphone 2.6% (257 data pairs) and smartwatch 1.8% (247 data pairs). Conclusions: Device SpO2 demonstrated RMSD < 3.0% to SaO2, meeting FDA/ISO clearance standards at the time of study. However, additional testing in persons with darker skin tones is necessary. Smartphones and paired wearables, when cleared for clinical use following revision of FDA clearance standards, may expand access to remote pulse oximetry.

Profound metabolomic alterations occur during COVID-19. Early identification of the subset of hospitalised COVID-19 patients at risk of developing severe disease is critical for optimal resource utilization and prompt treatment. This work explores the metabolomic profile of hospitalised adult COVID-19 patients with severe disease, and establishes a predictive signature for disease progression. Within 48 hours of admission, serum samples were collected from 148 hospitalised patients for nuclear magnetic resonance (NMR) spectroscopy. Lipoprotein profiling was performed using the 1H-NMR-based Liposcale test, while low molecular weight metabolites were analysed using one-dimensional Carr-Purcell-Meiboom-Gill pulse spectroscopy and an adaptation of the Dolphin method for lipophilic extracts. Severe COVID-19, per WHO's Clinical Progression Scale, was characterized by altered lipoprotein distribution, elevated signals of glyc-A and glyc-B, a shift towards a catabolic state with elevated levels of branched-chain amino acids, and accumulation of ketone bodies. Furthermore, COVID-19 patients initially presenting with moderate disease but progressing to severe stages exhibited a distinct metabolic signature. Our multivariate model demonstrated a cross-validated AUC of 0.82 and 72% predictive accuracy for severity progression. NMR spectroscopy-based metabolomic profiling enables the identification of moderate COVID-19 patients at risk of disease progression, aiding in resource allocation and early intervention.

The Coronavirus Disease 2019 (COVID-19), caused by virus SARS-CoV-2, is characterized by massive inflammation and immune system imbalance. Despite the implementation of vaccination protocols, the accessibility of treatment remains uneven. Furthermore, the persistent threat of new variants underscores the urgent need for expanded research into therapeutic options for SARS-CoV-2. Mesenchymal stem cells (MSCs) are known for their immunomodulatory potential through the release of molecules into the extracellular space, either as soluble elements or carried by extracellular vesicles (EVs). The aim of this study was to evaluate the anti-inflammatory potential of EVs obtained from human adipose tissue (ASC-EVs) against SARS-CoV-2 infection. ASC-EVs were purified by size-exclusion chromatography, and co-culture assays confirmed that ASC-EVs were internalized by human lung cells and could colocalize with SARS-CoV-2 into early and late endosomes. To determine the functionality of ASC-EVs, lung cells were infected with SARS-CoV-2 in the presence of increasing concentrations of ASC-EVs, and the release of cytokines, chemokines and viruses were measured. While SARS-CoV-2 replication was significantly reduced only at the highest concentrations tested, multiplex analysis highlighted that lower concentrations of ASC-EV sufficed to prevent the production of immune modulators. Importantly, ASC-EVs did not contain detectable inflammatory cytokines, nor did they trigger inflammatory mediators, nor affect cellular viability. In conclusion, this work suggests that ASC-EVs have the potential to attenuate inflammation by decreasing the production of pro-inflammatory cytokines in lung cells following SARS-CoV-2 infection.

Immune dysregulation and SARS-CoV-2 plasma viremia have been implicated in fatal COVID-19 disease. However, how these two factors interact to shape disease outcomes is unclear.

We carried out viral and immunological phenotyping on a prospective cohort of 280 patients with COVID-19 presenting to acute care hospitals in Boston, Massachusetts and Genoa, Italy between June 1, 2020 and February 8, 2022. Disease severity, mortality, plasma viremia, and immune dysregulation were assessed. A mouse model of lethal H1N1 influenza infection was used to analyze the therapeutic potential of Notch4 and pyroptosis inhibition in disease outcome.

Stratifying patients based on %Notch4+ Treg cells and/or the presence of plasma viremia identified four subgroups with different clinical trajectories and immune phenotypes. Patients with both high %Notch4+ Treg cells and viremia suffered the most disease severity and 90-day mortality compared to the other groups even after adjusting for baseline comorbidities. Increased Notch4 and plasma viremia impacted different arms of the immune response in SARS-CoV-2 infection. Increased Notch4 was associated with decreased Treg cell amphiregulin expression and suppressive function whereas plasma viremia was associated with increased monocyte cell pyroptosis. Combinatorial therapies using Notch4 blockade and pyroptosis inhibition induced stepwise protection against mortality in a mouse model of lethal H1N1 influenza infection.

The clinical trajectory and survival outcome in hospitalized patients with COVID-19 is predicated on two cardinal factors in disease pathogenesis: viremia and Notch4+ Treg cells. Intervention strategies aimed at resetting the immune dysregulation in COVID-19 by antagonizing Notch4 and pyroptosis may be effective in severe cases of viral lung infection.

Cepharanthine (CEP) is a natural remedy that potently inhibits SARS-CoV-2 activity both in vitro and in vivo. To evaluate the efficacy and safety of CEP compared with placebo in adults with asymptomatic or mild coronavirus disease 2019 (COVID-19), we conducted a proof-of-concept, double-blind, randomized, placebo-controlled trial. Patients were randomized to receive 120 mg/day of CEP, 60 mg/day CEP or placebo for 5 days. Main outcome was the time from randomization to negative nasopharyngeal swab and safety. Among 262 randomized participants, 188 completed the trial among group of 120 mg/day CEP (n = 65), 60 mg/day CEP (n = 68) and placebo (n = 55). Neither 120 mg/day or 60 mg/day CEP shortened the time to negative significantly compared with placebo. However, 60 mg/day CEP showed a slight trend (difference=-0.77 days, hazard ratio (HR) = 1.40, 95% CI 0.97-2.01, p = 0.072). In analysis of participants with good medication compliance, 60 mg/day CEP significantly shortened the time to negative (difference=-0.87 days, HR = 1.56, 95% CI 1.03-2.37, p = 0.035). Adverse events were not different among the three groups, and no serious adverse events occurred. In conclusion, treatment of asymptomatic or mild Covid-19 with 120 mg/day or 60 mg/day did not shorten the time to negative significantly. However, 60 mg/day CEP showed a slight trend which needs future confirmatory trials to validate. (NCT05398705).

Drivers of COVID-19 severity are multifactorial and include multidimensional and potentially interacting factors encompassing viral determinants and host-related factors (i.e., demographics, pre-existing conditions and/or genetics), thus complicating the prediction of clinical outcomes for different severe acute respiratory syndrome coronavirus (SARS-CoV-2) variants. Although millions of SARS-CoV-2 genomes have been publicly shared in global databases, linkages with detailed clinical data are scarce. Therefore, we aimed to establish a COVID-19 patient dataset with linked clinical and viral genomic data to then examine associations between SARS-CoV-2 genomic signatures and clinical disease phenotypes.

A cohort of adult patients with laboratory confirmed SARS-CoV-2 from 11 participating healthcare institutions in the Greater Toronto Area (GTA) were recruited from March 2020 to April 2022. Supervised machine learning (ML) models were developed to predict hospitalization using SARS-CoV-2 lineage-specific genomic signatures, patient demographics, symptoms, and pre-existing comorbidities. The relative importance of these features was then evaluated.

Complete clinical data and viral whole genome level information were obtained from 617 patients, 50.4% of whom were hospitalized. Notably, inpatients were older with a mean age of 66.67 years (SD ± 17.64 years), whereas outpatients had a mean age of 44.89 years (SD ± 16.00 years). SHapley Additive exPlanations (SHAP) analyses revealed that underlying vascular disease, underlying pulmonary disease, and fever were the most significant clinical features associated with hospitalization. In models built on the amino acid sequences of functional regions including spike, nucleocapsid, ORF3a, and ORF8 proteins, variants preceding the emergence of variants of concern (VOCs) or pre-VOC variants, were associated with hospitalization.

Viral genomic features have limited utility in predicting hospitalization across SARS-CoV-2 diversity. Combining clinical and viral genomic datasets provides perspective on patient specific and virus-related factors that impact COVID-19 disease severity. Overall, clinical features had greater discriminatory power than viral genomic features in predicting hospitalization.

The effects of aging on the organism manifest in various ways, including profound and complex changes in functioning patterns, responses to stimuli, and regenerative capacity. Nevertheless, it is remarkable that some elderly individuals maintain their health and functionality despite advanced age, showing resilience to environmental adversities, such as SARS-CoV-2 infection. In this study, we examined a unique cohort of 100 individuals older than 90 years, including centenarians, who recovered from COVID-19 before the availability of vaccines in Brazil. We performed whole-exome analyses and identified incidental findings in four participants. These findings included pathogenic variants associated with serious conditions, such as cancer predisposition and cardiovascular diseases. Specifically, variants were found in the RYR1, DSP, BRCA2, BRCA1, and TTN genes. Also, other two individuals were homozygous for rare variants in the TYK2 gene, related to primary immunodeficiencies. The significance of these findings is underscored by the fact that, despite carrying these rare variants, these individuals surpassed 90 years of age and survived the COVID-19 pandemic. This suggests the presence of genetic protective factors that contribute to longevity and resilience. Therefore, this study provides new insights into interpreting incidental findings in long-lived populations and raises important questions for clinical practice and the genetics of longevity.

Measuring virus in biofluids is complicated by confounding biomolecules coisolated with viral nucleic acids. To address this, we developed an affinity-based microfluidic device for specific capture of intact severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Our approach used an engineered angiotensin-converting enzyme 2 to capture intact virus from plasma and other complex biofluids. Our device leverages a staggered herringbone pattern, nanoparticle surface coating, and processing conditions to achieve detection of as few as 3 viral copies per milliliter. We further validated our microfluidic assay on 103 plasma, 36 saliva, and 29 stool samples collected from unique patients with COVID-19, showing SARS-CoV-2 detection in 72% of plasma samples. Longitudinal monitoring in the plasma revealed our device's capacity for ultrasensitive detection of active viral infections over time. Our technology can be adapted to target other viruses using relevant cell entry molecules for affinity capture. This versatility underscores the potential for widespread application in viral load monitoring and disease management.

Current evidence is inconsistent on whether vitamin D supplementation can prevent COVID-19 infection or improve its clinical outcomes. To better understand and look into the issue, we went through the background knowledge of COVID-19 and vitamin D, searched in Pubmed [by using key words in the title containing "randomized clinical trial", "COVID-19", and "vitamin D (25-hydroxyvitamin D, or cholecalciferol, or calcidiol, or calcifediol) supplementation"] for publications of studies on vitamin D/supplementation in COVID-19 patients, especially those about the randomized clinical trials (RCTs). After reviewing these papers, we did a short background review of vitamin D and the pathophysiology of COVID-19, summarized the key features of the 25 RCTs in text and tabulated in a table of some of the features, commented, compared and discussed the differences between RCTs (for example, change the serum 25-hydroxyvitamin D concentration from nmol/L to ng/mL, making the comparison easier). The take-home question of the review is that serum 25-hydroxyvitamin D concentration is an important indicator of the supplementation effect of vitamin D correction but may not be reliable in predicting the supplementation effect on the clinical outcomes of COVID-19.

Lymphopenia is recognized as a biomarker for predicting outcomes in coronavirus disease (COVID-19). However, the optimal timing for its observation remains uncertain. We investigated the association between early lymphopenia and COVID-19 prognosis, as well as the relationship between lymphocyte count trends and disease outcomes.

We analyzed data from the J-RECOVER study, a multicenter retrospective cohort study in Japan, encompassing patients with COVID-19 between January and September 2020. The patients were categorized into lymphopenia (LP) (<800 cells/μL) and non-lymphopenia (NL) (≥800 cells/μL) groups based on the lymphocyte counts between days 1 and 4 post-onset. They were further divided into "persistent," "recovered," "exacerbated," and "stable" groups based on lymphocyte counts between days 7 and 10. The primary outcome was the in-hospital mortality. The Cox proportional hazard regression was used for the analysis.

Of 995 enrolled patients, 212 patients (21.3%) were classified into the LP group. LP was significantly associated with in-hospital mortality (hazard ratio [HR] 2.32, [95% CI 1.39 to 3.87], p-value 0.001). In both the LP and NL groups, lower lymphocyte counts between 7 and 10 days-categorized as the "persistent" and "exacerbated" groups-was associated with in-hospital mortality (HR 4.65, [95% CI 2.07 to 10.47], p-value <0.001, and HR 5.59, [95% CI 2.24 to 13.97], p-value <0.001, respectively).

Early lymphopenia is predictive of poor prognosis in patients with COVID-19. A declining lymphocyte count trend post-onset further indicates disease deterioration.

Bruton's Tyrosine Kinase (BTK), an important element for the production of several inflammatory cytokines, may play a role in the pathogenesis of COVID-19. This study aimed to assess BTK gene expression levels in COVID-19 cases based on disease severity and outcome.

In this study, 33 hospitalized patients with COVID-19 were recruited and divided into two groups based on the severity of the disease: "mild to moderate" and "severe to critical". A blood sample was taken from each patient, peripheral blood mononuclear cells (PBMCs) were extracted, and BTK gene expression was measured. The level of BTK gene expression was compared based on the demographic data, laboratory results, and the severity and outcome of the disease.

Among the 33 patients, 22 (66.7%) were male, with nearly half having at least one underlying condition. The severity groups comprised 12 patients in the "mild to moderate" category and 21 in the "severe to critical" category, with eight (24.2%) experiencing fatal outcomes. Age, weight, and BMI showed no significant associations with BTK expression. BTK expression was notably lower in "severe to critical" and ICU-admitted cases, as well as in individuals with low O2 saturation. However, no significant difference in BTK expression was observed between cured and deceased patients (p = 0.117).

BTK gene expression in PBMCs exhibited an inverse correlation with COVID- 19 severity. However, no difference was found between BTK expression and disease outcome.

The severity of SARS-CoV-2 illness is influenced by factors including age, sex, pre-existing health conditions, and individual immune responses. However, the mechanisms conferring immunity following antigenic challenge have not been fully elucidated. There are currently no studies evaluating longitudinal proteomic changes in individuals following vaccination and breakthrough, limiting our understanding of the underlying mechanisms driving conferred immunity. In this work, we evaluated the differential protein expression in individuals with (CoV-P) or without (CoV-N) prior SARS-CoV-2 infection following primary vaccination and after breakthrough infection (CoV-BT). Overall, we found that individuals receiving primary vaccination relied on innate immune mechanisms, including complement and coagulation cascades, and natural killer cell-mediated cytotoxicity, while conversely, breakthrough infection immune mechanisms relied on T cell-mediated immunity. These mechanistic differences may help explain heterogeneity associated with vaccine-induced and breakthrough infection-related outcomes.

COVID-19, the disease caused by SARS-CoV-2, particularly causes severe inflammatory disease in elderly, obese, and male patients. Since both aging and obesity are associated with decreased testosterone and estradiol expression, we hypothesized that decreased hormone levels contribute to excessive inflammation in the context of COVID-19. Previously, we and others have shown that hyperinflammation in severe COVID-19 patients is induced by the production of pathogenic anti-spike IgG antibodies that activate alveolar macrophages. Therefore, we developed an in vitro assay in which we stimulated human macrophages with viral stimuli, anti-spike IgG immune complexes, and different sex hormones. Treatment with levels of testosterone reflecting young adults led to a significant reduction in TNF and IFN-γ production by human macrophages. In addition, estradiol significantly attenuated the production of a very broad panel of cytokines, including TNF, IL-1β, IL-6, IL-10, and IFN-γ. Both testosterone and estradiol reduced the expression of Fc gamma receptors IIa and III, the two main receptors responsible for anti-spike IgG-induced inflammation. Combined, these findings indicate that sex hormones reduce the inflammatory response of human alveolar macrophages to specific COVID-19-associated stimuli, thereby providing a potential immunological mechanism for the development of severe COVID-19 in both older male and female patients.

Transgelin is a central actin-binding protein of the calponin family and involved in the process of multiple pulmonary diseases. Nevertheless, the role of transgelin in Coronavirus disease 2019 (COVID-19) patients is confusing.

All 317 COVID-19 patients were recruited from two hospital. Peripheral blood was collected from the fasting patients at the onset and convalescent phases. Demographic data and clinical information were obtained. The expression of serum transgelin was estimated using ELISA.

The expression of serum transgelin on admission was gradually elevated in parallel with the increased severity scores of COVID-19. After treatment, serum transgelin expression was reduced during the convalescent phase. Spearman correlative analyses found that serum transgelin expression was closely correlated to lots of clinical parameters. Besides, serum transgelin was positively associated with severity scores. Follow-up research found that serum higher transgelin on admission elevated the risks of mechanical ventilation, vasoactive agent utilization, ICU admission, death, and longer hospital stays during hospitalization through a prospective cohort study. Additionally, there were similarly predictive capacities for critical patients and death between serum transgelin on admission and severity scores among COVID-19 patients.

The expression of serum transgelin is positively with the severity and poorly prognostic outcomes among COVID-19 patients, indicating that transgelin is implicated in the pathological process of COVID-19. Transgelin can assist in the risk stratification and revealing the pathological mechanisms of COVID-19.

The most appropriate anti-inflammatory treatment for moderate COVID-19 pneumonia remains uncertain. We aimed to compare the effectiveness of a high-dose methylprednisolone versus a high-dose dexamethasone in hospitalized moderate COVID-19 pneumonia, regarding the WHO clinical progression scales, mortality, and the length of hospitalization.

In this open-labeled randomized controlled trial, we enrolled patients with age > 18 years old who were diagnosed moderate COVID-19 pneumonia confirmed by real-time PCR, evidence of pneumonia by chest imaging and resting oxygen saturation between 90 and 94%. Patients were randomized at a 1:1 ratio to receive methylprednisolone 250 mg/day or dexamethasone 20 mg/day over the first three days. Then the patients in both groups received dexamethasone 20 mg/day on days 4-5, and 10 mg/day on days 6-10. Primary outcome was assessed by a 10-point WHO clinical progression scales ranging from uninfected (point 0) to death (point 10) on the fifth day of treatment. Secondary outcomes including 90-day mortality, length of hospitalization, rate of intensive care unit (ICU) transfer and complications were determined.

Of 98 eligible patients, the mean age was 76.0 ± 13.3 years. The median date of illness at the time of randomization was 3 days (interquartile range 2, 5). Baseline clinical characteristics and severity did not differ between groups. The WHO clinical progression scales were similar between methylprednisolone and dexamethasone group at 5 and 10 days of treatment [4.84, (95% confidence interval(CI), 4.35-5.33) vs. 4.76 (95% CI, 4.27-5.25), p = 0.821 and 4.32 (95% CI, 3.83-4.81) vs. 3.80 (95% CI, 3.31-4.29), p = 0.140, respectively)]. Both groups did not differ in-hospital mortality, length of hospitalization, and rate of ICU transfer. There were also no differences in steroid-related complications between groups until 90 days of follow-up.

In patients with moderate COVID-19 pneumonia, initial anti-inflammatory treatment with 250 mg/day of methylprednisolone for three days does not yield better outcomes over high-dose dexamethasone.

This study was registered at Thai Clinical Trials Registry on October 17, 2021, with the identifier TCTR20211017001.

In 2020, 20% of patients with COVID-19 developed severe complications, including life-threatening pneumonia with systemic inflammatory response syndrome (SIRS). We developed a preliminary SIRS monitor that does not require blood sampling, is noninvasive, and can collect data 24 h per day. The proposed monitor comprises a piezoelectric respiratory sensor located beneath the patient's mattress and a fingertip pulse sensor that determines ultra-high accuracy respiratory rate (mode of a 40-min frequency distribution of respiratory rates (M40FD-RR)). We assessed the clinical performance of the M40FD-RR preliminary SIRS monitor in 29 patients (12 female, 17 male, aged 15-90 years) hospitalized at Suwa Central Hospital with COVID-19, which was confirmed by a positive polymerase chain reaction test. SIRS was evaluated by logistic regression analysis using M40FD-RR, heart rate, age, and sex as explanatory variables. We compared the results of 109 examinations of 29 COVID-19 inpatients with SIRS against those determined by the proposed monitor. The proposed monitor achieved 75% sensitivity and 83% negative predictive value, making it a promising candidate for future 24 h noninvasive preliminary SIRS tests.

The long-term effects of SARS-CoV-2 infection, and their determinants, are still unknown. This study aimed to assess symptoms one year after admission for COVID-19, according to the organ/system involved, and to identify factors. Cross-sectional study with retrospective data collection from March 2020 to February 2021. Inclusion criteria: aged ≥ 18 years and admitted for COVID-19. Exclusion criteria: death, not localized, refusal to participate, cognitive impairment or language barrier. A telephone survey was conducted on long COVID-related symptoms one year after hospital discharge. n = 486. The most frequent symptom groups were neurological (n = 225; 46.3%) and respiratory (n = 201; 41.4%). Multivariable analysis showed that a history of anxiety was significantly associated with psychiatric symptoms (ORa = 2.04, 95%CI = 1.02-4.06), fibromyalgia/chronic fatigue with general symptoms (ORa = 11.59, 95%CI = 1.47-9.34) and obesity with respiratory (ORa 1.90, 95%CI = 1.27-2.83) and musculoskeletal symptoms (ORa 1.96, 95%CI = 1.30-2.96). Male sex was associated with a significantly lower risk of neurological (ORa 0.64, 95%CI = 0.44-0.93), respiratory (ORa 0.45, 95%CI = 0.31-0.67), general (ORa 0.43, 95%CI = 0.29-0.63), psychiatric (ORa 0.34, 95%CI = 0.22-0.51), musculoskeletal (ORa 0.47, 95%CI = 0.32-0.70), dermatological (ORa 0.24, 95%CI = 0.14-0.42) and digestive (ORa 0.38, 95%CI = 0.20-0.73) symptoms. Advanced age (≥ 71 years) also had a protective effect against general (ORa 0.60, 95%CI = 0.39-0.95), psychiatric (ORa 0.39, 95%CI = 0.23-0.64), and dermatological (ORa 0.47, 95%CI = 0.24-0.92) symptoms. Patients admitted for SARS-CoV-2 infection frequently experience symptoms at one year, especially neurological and respiratory symptoms. Female sex, obesity, a history of anxiety and fibromyalgia/chronic fatigue were independent risk factors for presenting symptoms. Advanced age acted as a protective factor.

It remains unclear whether pulmonary fibrosis-like changes differ in patients with different SARS-CoV-2 variants. This study aimed to compare pulmonary fibrotic changes between two SARS-CoV-2 variant periods (delta vs. pre-delta) in critically ill patients with SARS-CoV-2 pneumonia. Clinical data and chest CT images of patients with SARS-CoV-2 pneumonia receiving mechanical ventilation were collected from 10 hospitals in South Korea over two periods: delta (July-December, 2021; n = 64) and pre-delta (February, 2020-June, 2021; n = 120). Fibrotic changes on chest CT were evaluated through visual assessment. Of 184 patients, the mean age was 64.6 years, and 60.5% were ale. Fibrosis-like changes on chest CT (median 51 days from enrollment to follow up CT scan, interquartile range 27-76 days) were identified in 75.3%. Delta group showed more fibrosis-like changes (≥ 2) (69.8% vs. 43.1%, P = 0.001) and more frequent reticulation and architectural distortion+/-parenchymal band than pre-delta group. Even after propensity score matching with clinical variables, delta group had more severe (≥ 2) fibrosis-like changes (71.4% vs. 38.8%, P = 0.001), and more frequent reticulation and architectural distortion+/-parenchymal band than pre-delta group. Our data suggest that critically ill patients with SARS-CoV-2 in delta period had more severe pulmonary fibrosis-like changes than those in pre-delta period.

We investigated behavioral responses to COVID-19 in Malawi, where a first wave of the pandemic occurred between June and August 2020. Contrary to many countries on the African continent, the Government of Malawi did not impose a lockdown or a stay-at-home order in response to the initial spread of SARS-CoV-2. We hypothesized that, in the absence of such requirements to restrict social interactions, individuals would primarily seek to reduce the risk of SARS-CoV-2 transmission during contacts, rather than reduce the extent of their social contacts. We analyzed 4 rounds of a panel survey spanning time periods before, during and after the first wave of the COVID-19 pandemic in Malawi. Five hundred and forty-three participants completed 4 survey interviews between April and November 2020. We found that the likelihood of attending various places and events where individuals work and/or socialize remained largely unchanged during that time. Over the same time frame, however, participants reported adopting on a large scale several behaviors that reduce the transmissibility of SARS-CoV-2 during contacts. The percentage of panel participants who reported practicing physical distancing thus increased from 9.8% to 47.0% in rural areas between April-May 2020 and June-July 2020, and from 11.4% to 59.4% in urban areas. The percentage of respondents who reported wearing a facial mask to prevent the spread of SARS-CoV-2 also increased, reaching 67.7% among rural residents in August-September 2020, and 89.6% among urban residents. The pace at which these behaviors were adopted varied between population groups, with early adopters of mask use more commonly found among more educated office workers, residing in urban areas. The adoption of mask use was also initially slower among women, but later caught up with mask use among men. These findings stress the importance of behavioral changes in containing future SARS-CoV-2 outbreaks in settings where access to vaccination remains low. They also highlight the need for targeted outreach to members of socioeconomic groups in which the adoption of protective behaviors, such as mask use, might be delayed.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was responsible for causing the COVID-19 pandemic. Intensive research has illuminated the complex biology of SARS-CoV-2 and its continuous evolution during and after the COVID-19 pandemic. While much attention has been paid to the structure and functions of the viral spike protein and the entry step of viral infection, partly because these are targets for neutralizing antibodies and COVID-19 vaccines, the later stages of SARS-CoV-2 replication, including the assembly and egress of viral progenies, remain poorly characterized. This includes insight into how the activities of the viral structural proteins are orchestrated spatially and temporally, which cellular proteins are assimilated by the virus to assist viral assembly, and how SARS-CoV-2 counters and evades the cellular mechanisms antagonizing virus assembly. In addition to becoming infectious, SARS-CoV-2 progenies also need to survive the hostile innate and adaptive immune mechanisms, such as recognition by neutralizing antibodies. This review offers an updated summary of the roles of SARS-CoV-2 structural proteins in viral assembly, the regulation of assembly by viral and cellular factors, and the cellular mechanisms that restrict this process. Knowledge of these key events often reveals the vulnerabilities of SARS-CoV-2 and aids in the development of effective antiviral therapeutics.

This study aimed to describe the clinical course of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in patients with glomerular diseases (GDs) and its impact on the probability of relapse.

Patients with biopsy-proven GD and positive PCR test for SARS-CoV-2 from glomerular clinics across Greece were studied retrospectively. Those who received the GD diagnosis after the SARS-CoV-2 vaccination or coronavirus disease 2019 (COVID-19) or ended in ESKD prior to infection were excluded. Demographics, histopathological diagnoses, past medical history, immunosuppression, and GD activity status were recorded.

A total of 219 patients with GDs and documented SARS-CoV-2 infection were included. The mean time from the diagnostic kidney biopsy to SARS-CoV-2 infection was 67.6 ( ± 59.3) months. Among the participants, 82.5% had been vaccinated against SARS-CoV-2 with three doses (range: 2.5-3) without subsequent GD reactivation in 96.2% of them. Twenty-two patients (10%) were hospitalized for COVID-19 and one (0.5%) required mechanical ventilation. Four (1.8%) died due to COVID-19 and one (0.5%) had long COVID-19 symptoms. Among patients in remission prior to SARS-CoV-2 infection, 22 (11.2%) experienced a GD relapse within 2.2 (range: 1.5-3.7) months from the diagnostic test. The relapse-free survival after COVID-19 was significantly shorter for patients with minimal change disease, pauci-immune glomerulonephritis, and focal segmental glomerulosclerosis. No difference was observed in the relapse-free survival post-COVID-19 based on the history of SARS-CoV-2 vaccination.

SARS-CoV-2 infection appears to have a symptomatic but uncomplicated sequence in vaccinated patients with GDs, with a significant impact on the clinical course of GD, associated with an increased probability of relapse in certain histopathological types.

The persistent coronavirus disease 2019 (COVID-19) pandemic has had direct and indirect effects on mortality, making it essential to analyze excess mortality to fully understand the impact of the pandemic. In this study, we constructed a mathematical model using number of deaths from Statistics Korea and analyzed excess mortality between 2020 and 2022 according to age, sex, and dominant severe acute respiratory syndrome coronavirus 2 variant period.

Number of all-cause deaths between 2010 and 2022 were obtained from the annual cause-of-death statistics provided by Statistics Korea. COVID-19 mortality data were acquired from the Korea Disease Control and Prevention Agency. A multivariate linear regression model with seasonal effect, stratified by sex and age, was used to estimate the number of deaths in the absence of COVID-19. The estimated excess mortality rate was calculated.

Excess mortality was not significant between January 2020 and October 2021. However, it started to increase monthly from November 2021 and reached its highest point during the omicron-dominant period. Specifically, in March and April 2022, during the omicron BA.1/BA.2-dominant period, the estimated median values for excess mortality were the highest at 17,634 and 11,379, respectively. Both COVID-19-related deaths and excess mortality increased with age. A notable increase in excess mortality was observed in individuals aged ≥ 65 years. In the context of excess mortality per 100,000 population based on the estimated median values in March 2022, the highest numbers were found among males and females aged ≥ 85 years at 1,048 and 910, respectively.

This study revealed that the prolonged COVID-19 pandemic coupled with its high transmissibility not only increased COVID-19-related deaths but also had a significant impact on overall mortality rates, especially in the elderly. Therefore, it is crucial to concentrate healthcare resources and services on the elderly and ensure continued access to healthcare services during pandemics. Establishing an excess mortality monitoring system in the early stages of a pandemic is necessary to understand the impact of infectious diseases on mortality and effectively evaluate pandemic response policies.