Identifying plant-derived antiviral alkaloids as dual inhibitors of SARS-CoV-2 main protease and spike glycoprotein through computational screening.

COVID-19 is currently considered the ninth-deadliest pandemic, spreading through direct or indirect contact with infected individuals. It has imposed a consistent strain on both the financial and healthcare resources of many countries. To address this challenge, there is a pressing need for the development of new potential therapeutic agents for the treatment of this disease. To identify potential antiviral agents as novel dual inhibitors of SARS-CoV-2, we retrieved 404 alkaloids from 12 selected medicinal antiviral plants and virtually screened them against the renowned catalytic sites and favorable interacting residues of two essential proteins of SARS-CoV-2, namely, the main protease and spike glycoprotein. Based on docking scores, 12 metabolites with dual inhibitory potential were subjected to drug-likeness, bioactivity scores, and drug-like ability analyses. These analyses included the ligand-receptor stability and interactions at the potential active sites of target proteins, which were analyzed and confirmed through molecular dynamic simulations of the three lead metabolites. We also conducted a detailed binding free energy analysis of pivotal SARS-CoV-2 protein inhibitors using molecular mechanics techniques to reveal their interaction dynamics and stability. Overall, our results demonstrated that 12 alkaloids, namely, adouetine Y, evodiamide C, ergosine, hayatinine, (+)-homoaromoline, isatithioetherin C, N,alpha-L-rhamnopyranosyl vincosamide, pelosine, reserpine, toddalidimerine, toddayanis, and zanthocadinanine, are shortlisted as metabolites based on their interactions with target proteins. All 12 lead metabolites exhibited a higher unbound fraction and therefore greater distribution compared with the standards. Particularly, adouetine Y demonstrated high docking scores but exhibited a nonspontaneous binding profile. In contrast, ergosine and evodiamide C showed favorable binding interactions and superior stability in molecular dynamics simulations. Ergosine demonstrated exceptional performance in several key pharmaceutical metrics. Pharmacokinetic evaluations revealed that ergosine exhibited pronounced bioactivity, good absorption, and optimal bioavailability. Additionally, it was predicted not to cause skin sensitivity and was found to be non-hepatotoxic. Importantly, ergosine and evodiamide C emerged as superior drug candidates for dual inhibition of SARS-CoV-2 due to their strong binding affinity and drug-like ability, comparable to known inhibitors like N3 and molnupiravir. This study is limited by its in silico nature and demands the need for future in vitro and in vivo studies to confirm these findings.

Dynamic Assessment of Hematological Parameters as Predictive Biomarkers for Disease Severity and Prognosis in COVID-19 Patients: A Longitudinal Study.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to substantial morbidity and mortality worldwide. Hematological abnormalities are common in COVID-19 patients and play a significant role in disease pathogenesis and prognosis. OBJECTIVE: This study aimed to longitudinally monitor hematological parameters in COVID-19 patients and investigate their predictive value for disease severity and prognosis. METHODS: A prospective longitudinal design was employed to enroll 121 adult patients diagnosed with COVID-19 based on positive SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) test results. Baseline demographic and clinical data were collected, and hematological parameters, including complete blood count (CBC) indices, inflammatory markers, and coagulation profiles, were measured at predefined time points during hospitalization or outpatient visits. Follow-up assessments were conducted longitudinally to monitor the disease progression and clinical outcomes. RESULTS: This study revealed dynamic changes in hematological parameters over the course of COVID-19. Hemoglobin levels showed a decrease from baseline (mean ± SD: 12.5 ± 1.8 g/dL) to the peak of illness (10.2 ± 2.0 g/dL), indicating the development of anemia during the acute phase of infection. White blood cell counts demonstrated an initial increase (8.9 ± 3.2 × 10^9/L) followed by a decline (5.4 ± 1.9 × 10^9/L) as the disease progressed, suggesting an early inflammatory response followed by immune suppression. The platelet counts fluctuated, with a decrease observed during the acute phase (190 ± 50 × 10^9/L) and subsequent recovery during convalescence (240 ± 60 × 10^9/L). Inflammatory markers, such as C-reactive protein and interleukin-6, were elevated, peaking at 120 and 150 pg/mL, respectively, indicating systemic inflammation. Coagulation profiles showed abnormalities suggestive of COVID-19-associated coagulopathy, including elevated D-dimer levels (mean ± SD: 3.5 ± 1.2 µg/mL) and prolonged prothrombin time (15.8 ± 2.5 seconds). Longitudinal analysis of hematological parameters revealed associations between disease severity and clinical outcomes, with certain abnormalities correlating with an increased risk of complications and a poor prognosis. CONCLUSION: This study highlights the importance of monitoring hematological parameters in COVID-19 patients for risk stratification, prognostication, and guiding therapeutic interventions.

Management of patients with neurological diseases considering post-pandemic coronavirus disease 2019 (COVID-19) related risks and dangers - An updated European Academy of Neurology consensus statement.

BACKGROUND AND PURPOSE: In October 2020, the European Academy of Neurology (EAN) consensus statement for management of patients with neurological diseases during the coronavirus disease 2019 (COVID-19) pandemic was published. Due to important changes and developments that have happened since then, the need has arisen to critically reassess the original recommendations and address new challenges. METHODS: In step 1, the original items were critically reviewed by the EAN COVID-19 Task Force. In addition, new recommendations were defined. In step 2, an online survey with the recommendations forged in step 1 was sent to the Managing Groups of all Scientific and Coordinating Panels of EAN. In step 3, the final set of recommendations was made. RESULTS: In step 1, out of the original 36 recommendations, 18 were judged still relevant. They were edited to reflect the advances in knowledge and practice. In addition, 21 new recommendations were formulated to address the new knowledge and challenges. In step 2, out of the 39 recommendations sent for the survey, nine were approved as they were, whilst suggestions for improvement were given for the rest. In step 3, the recommendations were further edited, and some new items were formed to accommodate the participants' suggestions, resulting in a final set of 41 recommendations. CONCLUSION: This revision of the 2020 EAN Statement provides updated comprehensive and structured guidance on good clinical practice in people with neurological disease faced with SARS-CoV-2 infection. It now covers the issues from the more recent domains of COVID-19-related care, vaccine complications and post-COVID-19 conditions.

Kinetics of SARS-CoV-2 antibody titers after booster vaccinations during an Omicron surge in Japan.

BACKGROUND: Despite the emergence of SARS-CoV-2 variants and waning immunity after initial vaccination, data on antibody kinetics following booster doses, particularly those adapted to Omicron subvariants like XBB.1.5, remain limited. This study assesses the kinetics of anti-spike protein receptor-binding domain (S-RBD) IgG antibody titers post-booster vaccination in a Japanese population during the Omicron variant epidemic. METHODS: A prospective cohort study was conducted in Bizen City, Japan, from November 2023 to January 2024. Participants included residents and workers aged ≥18 years, with at least three COVID-19 vaccinations. Antibody levels were measured from venous blood samples. The study analyzed 424 participants and 821 antibody measurements, adjusting for variables such as age, sex, underlying conditions, and prior infection status. Mixed-effects models were employed to describe the kinetics of log-transformed S-RBD antibody titers. RESULTS: The study found that S-RBD antibody titers declined over time but increased with the number of booster vaccinations, particularly those adapted to Omicron and its subvariant XBB.1.5 (Pfizer-BioNTech Omicron-compatible: 0.156, 95%CI -0.032 to 0.344; Pfizer-BioNTech XBB-compatible: 0.226; 95%CI -0.051 to 0.504; Moderna Omicron-compatible: 0.279, 95%CI 0.012 to 0.546; and Moderna XBB-compatible: 0.338, 95%CI -0.052 to 0.728). Previously infected individuals maintained higher antibody titers, which declined more gradually compared to uninfected individuals (coefficient for interaction with time 0.006; 95%CI 0.001 to 0.011). Sensitivity analyses using Generalized Estimating Equations and interval-censored random intercept model confirmed the robustness of these findings. CONCLUSIONS: The study provides specific data on antibody kinetics post-booster vaccination, including the XBB.1.5-adapted vaccine, in a highly vaccinated Japanese population. The results highlight the importance of considering individual demographics and prior infection history in optimizing vaccination strategies.

Long-term longitudinal monitoring of SARS CoV-2 in urban rivers and sewers of Nepal.

In regions without adequate centralized wastewater treatment plants, sample collection from rivers and sewers can be an alternative sampling strategy for wastewater surveillance. This study aimed to assess the feasibility of alternative sampling strategies by testing samples collected from rivers (n = 246) and sewers (n = 244) in the Kathmandu Valley between March 2021 and February 2022. All samples were concentrated using the skimmed-milk flocculation method and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA was quantified using the nucleocapsid (N) and envelope (E) genes qPCR assays. Of the total, 75 % (371/490) of the samples tested positive using at least one qPCR assay, with concentrations ranging from 3.0 to 8.3 log10 gene copies/L. No significant correlation between concentrations of SARS-CoV-2 from both sewers and river with the number of confirmed coronavirus disease 2019 (COVID-19) cases in the Kathmandu valley was observed (p > 0.05). Despite the high concentration of SARS-CoV-2 in rivers and sewers, we hypothesize this finding to be a result of inaccurate number of clinical cases possibly due to inadequate clinical testing. This longitudinal study further supports the statement to consider sampling strategies from sewers and rivers for WBS in Nepal and other low and middle-income countries.

Non-invasive brain stimulation for post-COVID-19 conditions: a systematic review.

Alongside the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic, the number of patients with persistent symptoms following acute infection with SARS-CoV-2 is of concern. It is estimated that at least 65 million people worldwide meet criteria for what the World Health Organization (WHO) defines as "post-COVID-19 condition" - a multisystem disease comprising a wide range of symptoms. Effective treatments are lacking. In the present review, we aim to summarize the current evidence for the effectiveness of non-invasive or minimally invasive brain stimulation techniques in reducing symptoms of post-COVID-19. A total of nineteen studies were identified, one using transcutaneous vagus nerve stimulation (tVNS), another using transorbital alternating current stimulation (toACS), six studies on transcranial magnetic stimulation (TMS) and eleven studies on transcranial direct current stimulation (tDCS) for the treatment of post-COVID-19 symptoms. Existing studies report first promising results, illustrating improvement in clinical outcome parameters. Yet, the mechanistic understanding of post-COVID-19 and how brain stimulation techniques may be benefitial are limited. Directions for future research in the field are discussed.

Clusters of Emerging Multidrug-Resistant Organisms in United States Healthcare Facilities During the Initial Months of the SARS-CoV-2 Pandemic.

BACKGROUND: Outbreaks of emerging multidrug-resistant organisms (eMDROs), including carbapenem-resistant Enterobacterales (CRE), carbapenem-resistant Acinetobacter baumannii (CRAB), and Candida auris, have been reported among SARS-CoV-2 patients. We describe eMDRO clusters in SARS-CoV-2 units and associated infection control (IC) practices early in the SARS-CoV-2 pandemic. METHODS: We conducted a retrospective survey of a convenience sample of health departments in 11 states to describe clusters of eMDROs that began before November 1, 2020 and involved SARS-CoV-2 units. Cluster characteristics and IC practices during the cluster period were assessed using a standardized outbreak report form and descriptive analyses were performed. RESULTS: Overall, 18 eMDRO clusters (10 CRE, 6 C. auris, 1 carbapenem-resistant Pseudomonas aeruginosa, and 1 CRAB) in 18 healthcare facilities involving 397 patients were reported from 10 states. During the cluster period, 60% of facilities reported a shortage of isolation gowns, 69% extended use of gowns, and 67% reported difficulty obtaining preferred disinfectants. Reduced frequency of hand hygiene audits was reported in 85% of acute care hospitals during the cluster period compared to before the pandemic. CONCLUSIONS: Changes in IC practices and supply shortages were identified in facilities with eMDRO outbreaks during the SARS-CoV-2 pandemic and might have contributed to eMDRO transmission.

COVID-19 diarrhea is inflammatory, caused by direct viral effects plus major role of virus-induced cytokines.

BACKGROUND AND AIMS: Diarrhea occurs in up to 50% of cases of COVID-19. Nonetheless, the pathophysiologic mechanism(s) have not been determined. METHODS: This was examined using normal human enteroid monolayers exposed apically to live SARS-CoV-2 or non-replicating virus like particles (VLPs) bearing the four SARS-CoV-2 structural proteins or irradiated virus, all of which bound and entered enterocytes. RESULTS: Live virus and VLPs increased secretion of multiple cytokines and reduced mRNAs of ACE2, NHE3 and DRA. IL-6 plus IL-8 alone reduced NHE3 mRNA and protein and DRA mRNA. Neither VLPs nor IL-6 plus IL-8 alone altered Cl- secretion, but together they caused Cl- secretion, which was Ca2+ dependent, CFTR independent, blocked partially by a specific TMEM16 A inhibitor, and entirely by a general TMEM16 family inhibitor. VLPs and irradiated virus, but not IL-6 plus IL-8, produced Ca2+ waves that began within minutes of VLP exposure, lasted for at least 60 min, and were prevented by pretreatment with apyrase; a P2Y1 receptor antagonist; and general TMEM16 family inhibitor but NOT by the specific TMEM16A inhibitor. CONCLUSIONS: The pathophysiology of COVID-19 diarrhea appears to be a unique example of a calcium dependent inflammatory diarrhea, that is caused by direct viral effects plus the virus-induced intestinal epithelial cytokine secretion.

Extraction-Free Testing for SARS-CoV-2 in Nasal Swab and Saliva Samples on a Single High-Throughput Platform.

The COVID-19 pandemic introduced an urgent need for rapid and high-throughput testing for SARS-CoV-2. RNA extraction is a major bottleneck for RT-qPCR. We describe a semi-automated, extraction-free RT-qPCR assay for detection of SARS-CoV-2 in nasal swab and saliva samples on a single platform. With a limit of detection of 4 copies/mL, this laboratory developed test performed equivalently to established methods requiring nucleic acid extraction. Five technologists staffing two shifts per day (80 person-hours) processed more than 400,000 samples over 10 months. Patients opted to provide nasal swab samples (83.6%) more frequently than saliva (16.4%), creating the added challenge of producing swab collection kits. Real-world testing data indicated a higher frequency of SARS-CoV-2 detection in saliva (10.1%) compared to nasal swab (7.7%). This cost-effective and quickly scalable approach is suitable for pandemic preparedness planning related to surveillance and diagnostic testing.

Tetanus-diphtheria vaccine can prime SARS-CoV-2 cross-reactive T cells.

Vaccines containing tetanus-diphtheria antigens have been postulated to induce cross-reactive immunity to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which could protect against coronavirus disease (COVID-19). In this work, we investigated the capacity of Tetanus-diphtheria (Td) vaccine to prime existing T cell immunity to SARS-CoV-2. To that end, we first collected known SARS-CoV-2 specific CD8+ T cell epitopes targeted during the course of SARS-CoV-2 infection in humans and identified as potentially cross-reactive with Td vaccine those sharing similarity with tetanus-diphtheria vaccine antigens, as judged by Levenshtein edit distances (≤ 20% edits per epitope sequence). As a result, we selected 25 potentially cross-reactive SARS-CoV-2 specific CD8+ T cell epitopes with high population coverage that were assembled into a synthetic peptide pool (TDX pool). Using peripheral blood mononuclear cells, we first determined by intracellular IFNγ staining assays existing CD8+ T cell recall responses to the TDX pool and to other peptide pools, including overlapping peptide pools covering SARS-CoV-2 Spike protein and Nucleocapsid phosphoprotein (NP). In the studied subjects, CD8+ T cell recall responses to Spike and TDX peptide pools were dominant and comparable, while recall responses to NP peptide pool were less frequent and weaker. Subsequently, we studied responses to the same peptides using antigen-inexperienced naive T cells primed/stimulated in vitro with Td vaccine. Priming stimulations were carried out by co-culturing naive T cells with autologous irradiated peripheral mononuclear cells in the presence of Td vaccine, IL-2, IL-7 and IL-15. Interestingly, naive CD8+ T cells stimulated/primed with Td vaccine responded strongly and specifically to the TDX pool, not to other SARS-CoV-2 peptide pools. Finally, we show that Td-immunization of C57BL/6J mice elicited T cells cross-reactive with the TDX pool. Collectively, our findings support that tetanus-diphtheria vaccines can prime SARS-CoV-2 cross-reactive T cells and likely contribute to shape the T cell responses to the virus.

Associations between meteorological factors and COVID-19: a global scoping review.

Background: Many respiratory viruses and their associated diseases are sensitive to meteorological factors. For SARS-CoV-2 and COVID-19, evidence on this sensitivity is inconsistent. Understanding the influence of meteorological factors on SARS-CoV-2 transmission and COVID-19 epidemiology can help to improve pandemic preparedness. Objectives: This review aimed to examine the recent evidence about the relation between meteorological factors and SARS-CoV-2/COVID-19. Methods: We conducted a global scoping review of peer-reviewed studies published from January 2020 up to January 2023 about the associations between temperature, solar radiation, precipitation, humidity, wind speed, and atmospheric pressure and SARS-CoV-2/COVID-19. Results: From 9,156 initial records, we included 474 relevant studies. Experimental studies on SARS-CoV-2 provided consistent evidence that higher temperatures and solar radiation negatively affect virus viability. Studies on COVID-19 (epidemiology) were mostly observational and provided less consistent evidence. Several studies considered interactions between meteorological factors or other variables such as demographics or air pollution. None of the publications included all determinants holistically. Discussion: The association between short-term meteorological factors and SARS-CoV-2/COVID-19 dynamics is complex. Interactions between environmental and social components need further consideration. A more integrated research approach can provide valuable insights to predict the dynamics of respiratory viruses with pandemic potential.

Influence parental- and child-related factors on the acceptance of SARS-CoV-2 test methods in schools and daycare facilities.

Introduction: Rapid testing for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infections was an essential step in reducing the spread of the virus and monitoring pandemic development. Most mandatory standard pandemic testing in Germany has been performed in schools and daycare facilities. We investigated the influence of behavioral and attitudinal characteristics of children and caregivers on their acceptance of (i) antigen-based nasal swab rapid and (ii) oral saliva-based pooled Polymerase Chain Reaction (PCR) tests. Methods: Conducted through a cross-sectional survey between November and December 2021, with 1962 caregivers and 581 children/adolescents participating, the study evaluated the acceptability of each testing method on a six-point scale. Participants scored one test method conducted on their child at one of six levels with 1 and 6 denoting "excellent" (1) and "inadequate" (6), respectively. We considered demographic variables, vaccination status, child mental health (measured by the SDQ-questionnaire), and facility type (kindergarten, primary school, secondary school) as covariates. Results: Results reveal a preference for saliva-based PCR tests over nasal swabs by about one grade, particularly among parents of unvaccinated children, especially if their child expressed future vaccination reluctance. Testing acceptance was lower among children with mental health issues, primary school-aged, and those with less-educated parents. Perception of test accuracy and convenience influenced attitudes, favoring saliva-based PCR tests. Moreover, children with mental health issues felt less secure during testing. Discussion: To our knowledge, this is the first study to investigate the influence of different testing methods on testing acceptance for SARS-CoV-2 in children and caregivers. Our study identifies predictors of lower acceptance of public health surveillance measures and enables the development of educational programs on testing and vaccination tailored to the needs of specific target groups. Moreover, we demonstrate that test acceptance in vulnerable groups can be enhanced by careful choice of an appropriate testing method.

Effects of Recent Prior Dengue Infection on Risk and Severity of Subsequent SARS-CoV-2 Infection: A Retrospective Cohort Study.

Background and Aims: Elucidating whether prior dengue potentially confers cross-protection against COVID-19 is of public health importance in tropical countries at risk of overlapping dengue and COVID-19 epidemics. However, studies to date have yielded conflicting results. We aimed to assess effects of recent prior dengue infection on risk and severity of subsequent SARS-CoV-2 infection among adult Singaporeans. Methods: A retrospective cohort study including all adult Singaporeans aged ≥18 years was conducted from 1 July 2021 through 31 October 2022, when a dengue outbreak driven by the DENV3 serotype preceded subsequent waves of SARS-CoV-2 Delta/Omicron transmission in Singapore. SARS-CoV-2 and dengue infection status were classified using national registries. Cox regression models adjusted for demographics, COVID-19 vaccination status, comorbidity, and socioeconomic-status were used to assess risks and severity (hospitalization, severe illness) of SARS-CoV-2 infection occurring after previous recorded dengue infection. Results: A total of 3 366 399 individuals were included, contributing 1 399 696 530 person-days of observation. A total of 13 434 dengue infections and 1 253 520 subsequent SARS-CoV-2 infections were recorded; with an average of 94.7 days (standard deviation = 83.8) between dengue infection and SARS-CoV-2 infection. Preceding dengue infection was associated with a modest increase in risk of subsequent SARS-CoV-2 infection (adjusted hazards ratio [aHR] = 1.13; 95% confidence interval [CI], 1.08-1.17), and significantly elevated risk of subsequent COVID-19 hospitalization (aHR = 3.25; 95% CI, 2.78-3.82) and severe COVID-19 (aHR = 3.39; 95% CI, 2.29-5.03). Conclusions: Increased risk of SARS-CoV-2 infection and adverse COVID-19 outcomes were observed following preceding dengue infection in a national population-based cohort of adult Singaporeans. This observation is of significance in tropical countries with overlapping dengue and COVID-19 outbreaks.

Simultaneous endobronchial and mediastinal mucormycosis in a patient with Diabetes Mellitus and SARS-CoV-2: A case report and literature review.

Background: Mucormycosis can be lethal in people with immunocompromising conditions, especially Diabetes Mellitus. Correction of the underlying disorders, instant initiation of antifungal therapy, and surgical resection are the main components of treatment. Herin, we present the successful medical management of Mucormycosis in a patient with Diabetic Ketoacidosis and positive SARS-CoV-2 test who presented with a less seen condition: the simultaneity of mediastinal mass on one side and an endobronchial mass on the other. Case presentation: An 18-year-old male with a history of insulin-dependent DM from 4 years ago presented to our hospital with sudden onset dyspnea, chest pain, sore throat, hoarseness, cough, and sputum. Also, we detected unilateral swelling in the neck and multiple lymph nodes in the neck. Lung auscultation revealed bilateral generalized wheezing. Primary laboratory tests detected high blood sugar, metabolic acidosis, positive urine ketone, high ESR, positive CRP, and leukocytosis; his polymerase chain reaction (PCR) for SARS-CoV-2 was positive. Chest X-ray showed left upper lobe consolidation. Computed tomography scan (CT-scan) of the chest revealed a large collapse consolidation in the left lung, mild left side pleural effusion, mediastinal lymphadenopathy, and distention in the esophagus. With suspicion of malignancy, we performed flexible bronchoscopy and endobronchial Ultrasound (EBUS) which revealed a creamy tumoral lesion in the right main bronchus. The biopsy was consistent with Mucormycosis. We successfully treated Mucormycosis with Amphotericin-B liposomal. Conclusion: Mucormycosis can mimic the clinical characteristics of malignancy, and emphasize the importance of considering appropriate differential diagnoses because timely diagnosis and treatment is potentially life-saving in Mucormycosis.

COVID-19 vaccination and menstrual disturbances: A prospective study from Pakistan.

Objective: To evaluate whether or not immunization against COVID-19 is associated with changes in the duration and frequency of the menstrual cycle. Methods: This prospective analysis included the menstrual cycle data of 154 females after COVID-19 vaccination from August 2021 to March 2022. This study included Pakistani females aged 18 to 45 years and who had taken at-least one dose of COVID-19 vaccination. After two months of COVID vaccine the participants were interviewed again about the timing and duration of their menstrual cycle. The increase in menstrual length for >eight days was labelled as increased menstrual cycle duration. Results: Mean age of participants was 33.53±8.52 years. Among 154, 113 (73.4%) were married. Among 154 females, menstrual abnormality was reported by 59 (38.3%) females, increase in cycle duration was reported by 25 (16.2%) patients and decrease by 22 (14.3%), increase in number of bleeding days by 20 (13%) females and decrease by 15 (9.7%), increase in pain intensity was reported by 19 (12.3%) females and decrease by 17 (11.0%), increased intensity of blood flow was reported by 20 (13.0%) patients and decreased intensity by 19 (12.3%) females. Conclusion: COVID-19 vaccination is not associated with menstrual abnormalities in a significant number of females.

Functionalized N95 Face Mask with a Chemical-Free Paper-Based Collector for Exhaled Breath Analysis: SARS-CoV-2 Detection with a Printed Immunosensor as a Case Study.

Exhaled breath electrochemical sensing is a promising biomedical technology owing to its portability, painlessness, cost-effectiveness, and user-friendliness. Here, we present a novel approach for target analysis in exhaled breath by integrating a comfortable paper-based collector into an N95 face mask, providing a universal solution for analyzing several biomarkers. As a model analyte, we detected SARS-CoV-2 spike protein from the exhaled breath by sampling the target analyte into the collector, followed by its detection out of the N95 face mask using a magnetic bead-based electrochemical immunosensor. This approach was designed to avoid any contact between humans and the chemicals. To simulate human exhaled breath, untreated saliva samples were nebulized on the paper collector, revealing a detection limit of 1 ng/mL and a wide linear range of 3.7-10,000 ng/mL. Additionally, the developed immunosensor exhibited high selectivity toward the SARS-CoV-2 spike protein, compared to other airborne microorganisms, and the SARS-CoV-2 nucleocapsid protein. Accuracy assessments were conducted by analyzing the simulated breath samples spiked with varying concentrations of SARS-CoV-2 spike protein, resulting in satisfactory recovery values (ranging from 97 ± 4 to 118 ± 1%). Finally, the paper-based hybrid immunosensor was successfully applied for the detection of SARS-CoV-2 in real human exhaled breath samples. The position of the collector in the N95 mask was evaluated as well as the ability of this paper-based analytical tool to identify the positive patient.

Development of a Video Consultation Patient-Satisfaction Questionnaire (vCare-PSQ): A Cross-Sectional Explorative Study.

BACKGROUND: Since the COVID-19 pandemic, the use of video consultation (VC) in primary care has expanded considerably in many countries. VC and other telehealth formats are often touted as a solution to improved health care access, with numerous studies showing high satisfaction with this care format among health professionals and patients. However, operationalization and measurement of patient satisfaction with VC varies across studies and often lacks consideration of dynamic contextual factors (eg, convenience, ease-of-use, or privacy) and doctor-patient relational variables that may influence patient satisfaction. OBJECTIVE: We aim to develop a comprehensive and evidence-based questionnaire for assessing patient satisfaction with VC in general practice. METHODS: The vCare Patient-Satisfaction Questionnaire (the vCare-PSQ) was developed according to the COSMIN (Consensus-Based Standards for the Selection of Health Measurement Instruments) guidelines. To achieve our overall objective, we pursued three aims: (1) a validation analysis of an existing patient-satisfaction scale (the PS-14), (2) an assessment of extrinsic contextual factors that may impact patient satisfaction, and (3) an assessment of pertinent intrinsic and relational satisfaction correlates (eg, health anxiety, information technology literacy, trust in the general practitioner, or convenience). For validation purposes, the questionnaire was filled out by a convenience sample of 188 Danish adults who had attended at least 1 VC. RESULTS: Our validation analysis of the PS-14 in a Danish population produced reliable results, indicating that the PS-14 is an appropriate measure of patient satisfaction with VC in Danish patient populations. Regressing situational and doctor-patient relational factors onto patient satisfaction further suggested that patient satisfaction is contingent on several factors not measured by the PS-14. These include information technology literacy and patient trust in the general practitioner, as well as several contextual pros and cons. CONCLUSIONS: Supplementing the PS-14 with dynamic measures of situational and doctor-patient relational factors may provide a more comprehensive understanding of patient satisfaction with VC. The vCare-PSQ may thus contribute to an enhanced methodological approach to assessing patient satisfaction with VC. We hope that the vCare-PSQ format may be useful for future research and implementation efforts regarding VC in a general practice setting.

Immune signature of patients with cardiovascular disease predicts increased risk for a severe course of COVID-19.

Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection can lead to life-threatening clinical manifestations. Patients with cardiovascular disease (CVD) are at higher risk for severe courses of COVID-19. So far, however, there are hardly any strategies for predicting the course of SARS-CoV-2 infection in CVD patients at hospital admission. Thus, we investigated whether this prediction is achievable by prospectively analysing the blood immunophenotype of 94 nonvaccinated participants, including uninfected and acutely SARS-CoV-2-infected CVD patients and healthy donors, using a 36-colour spectral flow cytometry panel. Unsupervised data analysis revealed little differences between healthy donors and CVD patients, whereas the distribution of the cell populations changed dramatically in SARS-CoV-2-infected CVD patients. The latter had more mature NK cells, activated monocyte subsets, central memory CD4+ T cells, and plasmablasts but fewer dendritic cells, CD16+ monocytes, innate lymphoid cells, and CD8+ T-cell subsets. Moreover, we identified an immune signature characterised by CD161+ T cells, intermediate effector CD8+ T cells, and natural killer T (NKT) cells that is predictive for CVD patients with a severe course of COVID-19. Thus, intensified immunophenotype analyses can help identify patients at risk of severe COVID-19 at hospital admission, improving clinical outcomes through specific treatment.

Preconcentration and detection of SARS-CoV-2 in wastewater: A comprehensive review.

Severe acute respiratory syndrome coronaviruses 2 (SARS-CoV-2) causing coronavirus disease 2019 (COVID-19) affected the health of human beings and the global economy. The patients with SARS-CoV-2 infection had viral RNA or live infectious viruses in feces. Thus, the possible transmission of SARS-CoV-2 through wastewater received great attentions. Moreover, SARS-CoV-2 in wastewater can serve as an early indicator of the infection within communities. We summarized the preconcentration and detection technology of SARS-CoV-2 in wastewater aiming at the complex matrices of wastewater and low virus concentration and compared their performance characteristics. We described the emerging tests that would be possible to realize the rapid detection of SARS-CoV-2 in fields and encourage academics to advance their technologies beyond conception. We concluded with a brief discussion on the outlook for integrating preconcentration and the detection of SARS-CoV-2 with emerging technologies.

An overview of the development of pharmacotherapeutics targeting SARS-CoV-2.

Coronavirus disease 2019 (COVID-19) was declared a global pandemic in March 2020, which precipitated urgent public health responses. The causative agent, SARS-CoV-2, spreads primarily via respiratory droplets, necessitating precautions to mitigate transmission risks. Biopharmaceutical industries and academic institutions worldwide swiftly redirected their research endeavors towards developing therapeutic interventions, focusing on monoclonal antibodies, antiviral agents, and immunomodulatory therapies. The evolving body of evidence surrounding these treatments has prompted successive updates and revisions from the FDA, delineating the evolving landscape of COVID-19 therapeutics. This review comprehensively examines each treatment modality within the context of their developmental trajectories and regulatory approvals throughout the pandemic. Furthermore, it elucidates their mechanisms of action and presents clinical data underpinning their utility in combating the COVID-19 crisis.