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.

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.

The harms of promoting the lab leak hypothesis for SARS-CoV-2 origins without evidence.

Science is humanity's best insurance against threats from nature, but it is a fragile enterprise that must be nourished and protected. The preponderance of scientific evidence indicates a natural origin for SARS-CoV-2. Yet, the theory that SARS-CoV-2 was engineered in and escaped from a lab dominates media attention, even in the absence of strong evidence. We discuss how the resulting anti-science movement puts the research community, scientific research, and pandemic preparedness at risk.

Unlocking Secrets: Lipid Metabolism and Lipid Droplet Crucial Roles in SARS-CoV-2 Infection and the Immune Response.

Lipid droplets (LD) are crucial for maintaining lipid and energy homeostasis within cells. LDs are highly dynamic organelles that present a phospholipid monolayer rich in neutral lipids. Additionally, LDs are associated with structural and non-structural proteins, rapidly mobilizing lipids for various biological processes. Lipids play a pivotal role during viral infection, participating during viral membrane fusion, viral replication, and assembly, endocytosis, and exocytosis. Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) infection often induces LD accumulation, which is used as a source of energy for the replicative process. These findings suggest that LDs are a hallmark of viral infection, including SARS-CoV-2 infection. Moreover, LD participates in the inflammatory process and cell signaling, activating pathways related to innate immunity and cell death. Accumulating evidence demonstrates that LD induction by SARS-CoV-2 is a highly coordinated process, aiding replication and evading the immune system, and may contribute to the different cell death process observed in various studies. Nevertheless, recent research in the field of LDs suggests these organelles according to the pathogen and infection conditions may also play roles in immune and inflammatory responses, protecting the host against viral infection. Understanding how SARS-CoV-2 influences LD biogenesis is crucial for developing novel drugs or repurposing existing ones. By targeting host lipid metabolic pathways exploited by the virus, it is possible to impact viral replication and inflammatory responses. This review seeks to discuss and analyze the role of LDs during SARS-CoV-2 infection, specifically emphasizing their involvement in viral replication and the inflammatory response.

Mild coronavirus disease 2019: Results of a multicentric prospective study on severe acute respiratory syndrome coronavirus 2 presence in semen fractions and cells and its effects on semen parameters.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2, the virus responsible for coronavirus disease 2019, affects multiple organs. The virus enters cells through angiotensin-converting enzyme-2 and host factors present in genital organs, leading to concern over virus shedding in semen and reproductive function. OBJECTIVES: To investigate severe acute respiratory syndrome coronavirus 2 in semen from patients with a mild infection, identify the seminal infected cells, and explore the effect of the infection on sex hormones and semen parameters. MATERIALS AND METHODS: Prospective study of 54 men with mild severe acute respiratory syndrome coronavirus 2 infection. Semen was collected at 7, 15, 30, 60, 90, 180, and 365 days after symptom onset, and severe acute respiratory syndrome coronavirus 2 RNA was measured in serum, saliva, urine, and semen. The presence of infectious severe acute respiratory syndrome coronavirus 2 in semen was assessed using Vero cell culture. Infected semen cells were identified using immunofluorescence against severe acute respiratory syndrome coronavirus 2 nucleoprotein antigen and cell markers. Semen characteristics as well as testosterone, inhibin B, luteinizing hormone, and follicle-stimulating hormone levels were determined. RESULTS: 11% of patients had at least one severe acute respiratory syndrome coronavirus 2 RNA-positive semen. One patient had viral semen shedding up to day 90 after infection onset, with replication-competent virus isolated from semen and 40% cell fraction at day 7. After sperm preparation, 90% fraction was severe acute respiratory syndrome coronavirus 2 RNA-positive at days 7 and 15. The swim-up fraction was positive only on day 7. In semen, nucleoprotein antigen was detected mainly in exfoliated epithelial cells and less frequently in Sertoli cells. Sperm count and motile sperm count were lower at day 30 than at day 7. Round cells in semen were increased during the acute phase. At days 7 and 15, sperm count and motile sperm count were lower in severe acute respiratory syndrome coronavirus 2 RNA-positive semen compared with negative semen, while semen volume and follicle-stimulating hormone levels were increased. Long-term follow-up shows no evidence of a detrimental effect on hormonal or semen characteristics. DISCUSSION AND CONCLUSION: 11% of patients with mild coronavirus disease 2019 who were not hospitalized had severe acute respiratory syndrome coronavirus 2 excretions in semen, which persisted for up to 90 days in one patient. No germ cells appeared infected by the virus, but the detection of nucleoprotein antigen-positive epithelial semen cells and Sertoli cells suggests genital tract infection. Albeit infrequent, semen may contain the replication-competent virus during the acute phase with potential risk of severe acute respiratory syndrome coronavirus 2 transmissions during sexual contact and assisted reproduction procedures. The effect of mild coronavirus disease 2019 on spermatogenesis and reproductive hormones was moderate and reversible.

Postoperative Outcomes in Surgical Coronavirus Disease 2019 Patients: A Retrospective Cohort Study.

PURPOSE: Patients infected with severe acute respiratory syndrome coronavirus-2 (SARS-COV-2) who require surgical procedures are likely to experience higher postoperative mortality and morbidity. Our objective was to evaluate the perioperative course of patients infected with SARS-COV-2 undergoing surgical procedures. The purpose of this study was to describe the characteristics, outcomes, and the effect of the presence of symptoms. DESIGN: Retrospective cohort. METHODS: We analyzed the records of patients with SARS-CoV-2 infection who underwent surgical procedures from March 2020 to March 2021. Patients with ongoing infection at the time of surgery and those who had recently recovered were included. The primary outcome measure was 30-day in-hospital mortality after surgery. Secondary outcomes were intensive care unit (ICU) admission, length of stay in ICU, postoperative length of stay, and complications. FINDINGS: Data from 102 patients were analyzed. Twenty-four patients (23.5%) died postoperatively in the hospital within 30 days. Forty-four patients required ICU admission (average stay 13 days). The median postoperative length of stay was 8 days (interquartile range, 3.75 to 19.25 days). Pulmonary, thromboembolic, and surgical complications were noted in 29 (28.4%), 14 (13.7%), and 18 (17.6%), respectively. Patients aged 41 to 60 years experienced higher rates of pulmonary and thromboembolic complications. Comparison of asymptomatic versus symptomatic patients revealed significantly higher 30-day in-hospital mortality (9 [15%] vs 15 [35.7%], P = .019), ICU admission (17 [28.3%] vs 27 [64.3%], P < .001), length of stay in ICU (3 [2 to 11.5] vs 18 [7 to 27], P = .001), postoperative length of stay (6 [3 to 10.75] vs 12 [5 to 25.25], P = .016) and pulmonary complication rates (11 [18.3%] vs 18 [42.9%], P = .008) in the symptomatic patients. CONCLUSIONS: Symptomatic SARS-COV-2 patients undergoing surgical procedures experience significantly higher 30-day in-hospital mortality, ICU admission, longer ICU and hospital stay, and pulmonary complications.

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.

Paxlovid reduces the 28-day mortality of patients with COVID-19: a retrospective cohort study.

PURPOSE: In this study, we aim to explore the efficacy of paxlovid on reducing mortality of COVID-19 patients in clinical setting, especially whether paxlovid modifies the risk of death in these severe and critical patients. METHODS: Our retrospective cohort study was conducted on the medical records of patients, consecutively admitted for COVID-19 to five hospitals in Chongqing, China from Dec 8, 2022 to Jan 20, 2023. Based on whether patients received paxlovid during their hospitalization, patients were grouped as paxlovid group and non-paxlovid group. We used 1:1 ratio propensity score matching (PSM) in our study to adjust for confounding factors and differences between groups. Statistical analysis were performed by SPSS 23.0. The differences in 28-day mortality between these two groups and its influencing factors were the main results we focused on. RESULTS: There were 1018 patients included in our study cohort. With 1:1 ratio PSM, each of the paxlovid group and non-paxlovid group included 237 patients. The results showed that patients using paxlovid have a lower 28-day mortality in overall population either before PSM (OR 0.594, 95% CI 0.385-0.917, p = 0.019) or after PSM (OR 0.458, 95% CI 0.272-0.774, p = 0.003) with multivariable adjusted logistic regression models. Meanwhile, in severe subgroup, it showed similar findings.With paxlovid treatment, it showed a significantly lower 28-day mortality in severe subgroup both before PSM (28% vs.41%, p = 0.008) and after PSM (19% vs.32%, p = 0.007). CONCLUSION: Paxlovid can significantly reduce the risk of 28-day mortality in overall population and severe subgroup patients.This study distinguished the severe subgroup patients with COVID-19 who benefit more from paxlovid treatment.

4-(3-Phenylsulfonylindol-2-yl)-1-(pyridin-2-yl)piperazinyl-methanones as Potent Inhibitors of both SARS-CoV-2 and HCoV-OC43 Viruses.

SARS-CoV-2 and HCoV-OC43 belong to the same ß genus of the Coronaviridae family. SARS-CoV-2 was responsible for the recent COVID-19 pandemic, and HCoV-OC43 is the etiological agent of mild upper respiratory tract infections. SARS-COV-2 and HCoV-OC43 co-infections were found in children with respiratory symptoms during the COVID-19 pandemic. The two ß-coronaviruses share a high degree of homology between the 3CLpro active sites, so much so that the safer HCoV-OC43 has been suggested as a tool for the identification of new anti-SARS-COV-2 agents. Compounds 5 and 24 inhibited effectively both Wuhan and British SARS-CoV-2 patient isolates in Vero E6 cells and the HCoV-OC43 in MRC-5 cells at low micromolar concentrations. The inhibition was apparently exerted via targeting the 3CLpro active sites of both viruses. Compounds 5 and 24 at 100 µM inhibited the SARS-CoV-2 3CLpro activity of 61.78 and 67.30%, respectively. These findings highlight 5 and 24 as lead compounds of a novel class of antiviral agents with the potential to treat SARS-COV-2 and HCoV-OC43 infections.

Should SARS-CoV-2 serological testing be used in the decision to deliver a COVID-19 vaccine booster? A pro-con assessment.

Anti-SARS-CoV-2 vaccination has saved millions of lives in the past few years. To maintain a high level of protection, particularly in at-risk populations, booster doses are recommended to counter the waning of circulating antibody levels over time and the continuous emergence of immune escape variants of concern (VOCs). As anti-spike serology is now widely available, it may be considered a useful tool to identify individuals needing an additional vaccine dose, i.e., to screen certain populations to identify those whose plasma antibody levels are too low to provide protection. However, no recommendations are currently available on this topic. We reviewed the relevant supporting and opposing arguments, including areas of uncertainty, and concluded that in most populations, spike serology should not be used to decide about the administration of a booster dose. The main counterarguments are as follows: correlates of protection are imperfectly characterised, essentially owing to the emergence of VOCs; spike serology has an intrinsic inability to comprehensively reflect the whole immune memory; and booster vaccines are now VOC-adapted, while the commonly available commercial serological assays explore antibodies against the original virus.

Clinical feature of omicron infection in children with inborn errors of immunity in China.

Introduction: SARS-CoV-2 infection is hypothesized to be more severe in immunocompromised patients; however, clinical outcomes in children with inborn errors of immunity (IEI) during the Omicron pandemic in China have not been reported. Methods: This cohort study retrospectively reviewed 71 SARS-CoV-2-infected children with IEI using nationwide data from the National Center for Children's Health of China. COVID-19 was diagnosed by a positive rapid antigen or nucleic acid test result. Results: Among 71 SARS-CoV-2-infected children with IEI, male preponderance (male: female ratio of ~1.8:1), a median age of 8 years (IQR 3-11), and a predominance of antibody deficiency (19/71, 26.8%) were detected. Most of the patients got infected through household transmission, while a small proportion of them did so during hospital visits. The mean time periods were 3.3 days (n=44) for incubation, 8.4 days for symptoms (n=69), and 8.8 days for viral shedding (n=37). The time to viral shedding was proportional to the symptomatic period (R2 = 0.1243, p=0.0323) and prolonged in children with X- linked agammaglobulinemia. The most common symptoms of COVID-19 were fever, and some children showed only aggravation of the underlying disease. 15% of IEI children progress to pneumonia, 85% require medication, 17% are admitted to hospital, and 4.1% are classified as critical. Previously application of anti- infective medications was associated with an increased risk of hospitalization after COVID-19 infection. Of the 71 children with IEI, all recovered from COVID- 19. Conclusion: Overall, Omicron variant did not cause significant life-threatening infections among children with IEI in China, and most of them had a good clinical outcome. Nevertheless, these children exhibit an increased vulnerability to higher hospitalization rates, pneumonia, and severe illness compared to the general pediatric population.

Clinical characteristics of COVID-19 in children: a large multicenter study from Iran.

Introduction: During the COVID-19 pandemic, pediatric cases typically exhibit milder symptoms and lower mortality rates. However, the delay in pediatric vaccination poses major risks for children. This multicenter study aimed to comprehensively analyze demographic characteristics, clinical features, disease severity, and risk factors for ICU admission in Iran. Materials and methods: This observational study enrolled children aged 0-21 years with confirmed or probable COVID-19 diagnoses, referred from selected hospitals across 17 counties in Mazandaran province, Iran, between February 19 and August 14, 2021. Patients were categorized into mild, moderate, severe, or critical cases based on clinical and radiographic criteria. Data were extracted from medical records and analyzed using statistical methods. Logistic regression analysis was performed to identify factors associated with ICU admission and disease severity. Results: Among the 1,031 children included in the study, 61 were diagnosed with MIS-C. The distribution of patients by severity was 156 mild, 671 moderate, and 204 severe/critical cases. Age distribution significantly differed across severity groups (P < 0.001), with 55.19% aged over 5 years and 54% being male. 11.44% had underlying diseases. Fever (71.97%) was the most common symptom, followed by cough (34.43%) and dyspnea (24.83%). Within the inpatient group, 19.77% required ICU admission, with 0.91% mortality, including 3 MIS-C cases. Children with underlying diseases, gastrointestinal symptoms, and obesity had 4.16, 3.10-, and 2.17-times higher likelihood of ICU admission, respectively. Conclusion: Our study emphasized the importance of recognizing pediatric COVID-19 severity and symptoms. While fever, cough, and dyspnea prevailed, mortality rates were relatively low. However, comorbidities, obesity, and gastrointestinal symptoms linked to ICU admission, stressing early intervention. BMI also impacted disease severity and admission rate. Vaccination and targeted interventions are essential for protecting vulnerable children and easing healthcare strain.