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Background: Resident memory T cells (TRM) present at the respiratory tract may be essential to enhance early SARS-CoV-2 viral clearance, thus limiting viral infection and disease. While long-term antigen-specific TRM are detectable beyond 11 months in the lung of convalescent COVID-19 patients after mild and severe infection, it is unknown if mRNA vaccination encoding for the SARS-CoV-2 S-protein can induce this frontline protection. Method(s): We obtained cross-sectional paired blood and lung biopsy samples from patients (n=30) undergoing lung resection for various reasons and assigned them to one of four groups: I.) uninfected unvaccinated individuals (n=5), II.) unvaccinated long-term SARS-CoV-2 convalescent individuals (between 6.0-10.5 months post-infection;n=9), III.) uninfected and long-term vaccinated individuals (between 6.0-7.7 months after the second or third dose;n=10), and IV.) uninfected and short-term vaccinated individuals (between 1.3-1.8 months after the third or fourth dose;n=6). We determined the presence of SARS-CoV-2-specific CD4+ and CD8+ T cells in blood and lung samples after exposure of cells to M, N, and S peptide pools, followed by flow cytometry to detect TRM cells expressing interferon (IFN)gamma and/or CD107a, as a degranulation marker. Result(s): We found that the frequency of CD4+ T cells secreting IFNgamma in response to S-peptides was variable but detectable in blood and lung up to 8 months after mRNA vaccination. Moreover, the IFNgamma response of CD4+ T cells in the lung of mRNA-vaccinated patients was similar to the response found in convalescent patients. However, in mRNA-vaccinated patients, lung responses presented less frequently with a TRM phenotype compared to convalescent infected individuals and, strikingly, polyfunctional CD107a+ IFNgamma+ TRM were virtually absent in vaccinated patients. Conclusion(s): mRNA vaccines might induce memory responses within the lung parenchyma in some patients, potentially contributing to the overall disease control. However, the robust and broad TRM response established in convalescent-infected individuals may offer advantages at limiting disease if the virus is not blocked by initial mechanisms of protection, such as neutralization. Our results warrant investigation of mucosal vaccine-induced resident T cell responses in establishing superior site-specific protective immunity.
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Due to the SARS-COV-2 pandemic, educational institutions are immediately faced with a new challenge to adapt, forcing the transition from face-to-face teaching to distance learning in a short period. Distance education supported by technology is a challenge for educational institutions based on binomial technology/teaching. This paper presents a proposal for an e-learning technology structure, supported by a cluster of servers capable of responding to the requirements of distance learning based on the premises of High Availability, High Performance, Load Balancing. The beginning of this study consisted of a literature review to find the various existing technologies, a way to combine them and create a system capable of providing the necessary functionalities, and whose performance could host all the users of an institution simultaneously. The implemented system results from this combination of technologies and allows its capacity to be scaled at any moment according to momentary needs. In technological terms, the solution was based on a free Linux distribution, the Ubuntu Server installed inside a cluster of servers with VMware ESXi, and a cluster of database nodes based on Gallera technology. The eLearning platform used in this study was Moodle because it is one of the resources most used by institutions. The aspects of teaching, provision of content and execution of evaluation tests, were explored. With the implementation of the presented scenario, it was possible to guarantee the High Availability and load balancing of the platform and guarantee a high performance of the whole solution. © 2022 IEEE.
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This article demonstrates the validity and reliability of an instrument to evaluate the level of digital competence of Early Childhood Education and Primary Education teachers to carry out online tutorials using ICT resources as part of the educational process during the COVID-19 pandemic. The instrument consists of 35 items classified into five dimensions (A - functions of the tutor with the students;B - functions of the tutor with the teaching staff;C - functions of the tutor with the family;D - training in ICT and transfer;E - use of ICT resources). The instrument was applied to a sample of 1,098 active teachers from 14 autonomous communities located in Spain. Reliability was measured using Cronbach's Alpha, the SpearmanBrown Coefficient, Guttman Two Halves, Omega McDonald, and for composite reliability. To check the validity of the instrument, we analysed the validity of understanding, and exploration of dimensionality using Factorial-Exploratory Analysis (EFA), and the instrument was adjusted for the different models through Confirmatory Factor Analysis (CFA). In addition, factorial invariance was evaluated based on the variables sex (male/female), type of centre (public/charter school), and type of teaching staff (Early Childhood Education/Primary Education), as well as external validity. The results of the reliability analyses were highly satisfactory and, in relation to the construct validity, the results found a good fit of the model, both in the internal validity and in the factorial invariance. The final version of the instrument consists of 25 items.
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Background and objectives: The COVID-19 pandemic situation has led to severe restrictions, including mandatory home confinement. Sport as a means to reduce stress and psychological discomfort results in the following research that aims to compare the state of psychological discomfort and stress during this period of confinement, of a group of people, based on their level of sporting activity and sex. Material and Method: we use a descriptive, cross-sectional design;A relational study is developed to analyze the degree of dependence between the variables sex and level of sports practice with stress and psychological discomfort, for this we have used the scale of perceived stress and that of psychological discomfort of Kessler, with a sample composed of a total of 660 participants (301 women [45.6%] and 359 men [54.4%]). Results: Significant differences were found in the study groups according to the level of sports practice, as well as their sex, referring to psychological discomfort and stress levels. Conclusions: It is concluded that confinement during the pandemic is associated with significantly high levels of psychological distress and stress that can reach the threshold of clinical relevance. Lower scores at higher athletic level.
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Background: In order to inform vaccine development on the correlates of protection against SARS-CoV-2, we performed detailed phenotypic and functional analyses in clinically-defined groups of patients recruited during the first wave of SARS-CoV-2 infection, including the assessment of Resident Memory T cells (TRM) in lung of convalescent patients. Methods: Blood samples from 46 participants diagnosed with acute COVID-19 (14 symptomatic non-hospitalized;20 mild-hospitalized and 12 severehospitalized) were obtained 7-16 days after symptoms onset. Lung biopsies were obtained from three convalescent patients 1 to 7.5 months after initial infection. The phenotype and functional capabilities of SARS-CoV-2-specific CD4+ and CD8+T cells were measured by FACS after stimulation with a pool of overlapping SARS-CoV-2 viral peptides (M, N and S). Results: Pattern variations associated with viral-specific T cell responses where based on two factors, the targeted viral protein and the cohort of patients assessed. Overall, stimulation with M and N viral peptides induced a Th1 profile exemplified by IFNg production in CD4+T cells and degranulation in CD8+T cells respectively, whereas S peptides induced a Th2 profile exemplified by IL-4. Hospitalized patients showed increased IFNg secretion in CD4+T cells in response to any viral protein compared to non-hospitalized patients (p=0.020 for M and S peptides in the mild group;p=0.004 for M, p=0.011 for N and p=0.007 for S peptides in the severe group;Figure 1) and IL-4 secretion in CD8+T cells in response to S peptides (p=0.004 and p=0.003 for mild and severe patients, respectively). In contrast, the expression of IL-10, which was mostly expressed in CCR7+ cells, was significantly increased in CD4+T cells from non-hospitalized patients after stimulation with M peptides when compared to the mild COVID-19 group (p=0.035). Importantly, SARS-CoV-2 specific T cell responses with a biased TRM profile were detected up to 7.5 months after infection in the lung of convalescent patients. However, tissue responses strongly differed from blood. Conclusion: Our results suggest that a balanced anti-inflammatory antiviral response promoted by non-spike proteins may be key to favor infection resolution without major complications. Further, while immune responses migrate and establish in the lung as resident memory T cells, the magnitude and profile of the lung SARS-Cov-2 specific T cells strongly differ from the response detected in blood.
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Background: No effective drugs against SARS-CoV-2 infection are available. Screening of therapeutic candidates is primarily performed using immortalized cell lines. However, primary cell targets might show intrinsic differences in the expression profile of relevant host proteins, required for viral replication that could significantly affect the activity and potency of antivirals. Thus, the development of more physiological models for antiviral drug screening are urgently needed. Methods: Lung tissue was obtained from routinely thoracic surgical resections and was immediately digested before experiment set up. Cell populations and expression of ACE2 were characterized by FACS, and cell targets for SARS-CoV-2 were identified using a VSV∗ΔG(GFP)-S pseudotyped virus. 39 repurposing drugs previously identified by in silico models as potential viral entry inhibitors were tested using a VSV∗ΔG(Luc)-S virus. Cytotoxic concentration (CC50) and inhibitory concentration (IC5O) values were calculated using a non-linear regression dose-response curve and were compared to drug activity in VeroE6 cells. Results: Alveolar type II (AT-II) cells, the main cell target for SARS-CoV-2 infection in lungs, were identified within a fraction of cells characterized by CD45-, CD31-, EpCAM+ and HLA-DR+, (∼0.01-0.5% of viable cells). Using a VSV∗ΔG(GFP)-S virus we showed that viral entry was occurring in cells compatible with an AT-II phenotype, and infection was efficiently blocked with an anti-ACE2 antibody (Figure 1). Despite low and variable numbers of AT-II targets, antiviral assays using VSV∗ΔG(Luc)-S were highly sensitive and reproducible (CV of 17%). Compared with VeroE6 cells, IC50 values trended to be higher in tissues. Moreover, we found that 12.8% of the tested compounds had discordant results, where 10.25% of the drugs showed some antiviral effect in lung cell suspensions but no activity in VeroE6 and 3.9% showed only antiviral effect in VeroE6. Modulation of ACE2 expression by some of these compounds was also highly discordant between the cell line and lung tissue. Cepharantine (IC50=6μM, CC50=14μM) and Ergoloid (IC50=4.3μM, CC50=24μM) were identified as the most active entry inhibitors in lung cell suspension. Conclusion: The use of lung tissue for the screening of antiviral compounds represents a valid physiological and relevant model, which evidences intrinsic discrepancies with cell lines. Importantly, we identified repurposing drugs against SARS-CoV-2 with potential for clinical testing.
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Background: Out-of-Hospital Cardiac Arrest (OHCA) is the third leading cause of death in developed countries, with a survival rate less than 10%. Actually, although 70% of OHCA episodes are witnessed by laypeople only a third receive Basic Life Support (BLS). In Portugal, bystander-performed Cardiopulmonary Resuscitation (CPR) rate is still below 17%, with a survival rate of 4%. One of the most effective ways to increase bystander CPR is by training school-age children in BLS and Automated External Defibrillator (AED). Aims: Evaluate the capacity to learn and the quality of BLS maneuvers performed by school-age children. Methods: During a four-month period (interruption due to SARS-CoV-2 pandemic), BLS-AED teaching sessions were provided across schools in Oeiras, Portugal. It is considered that children older than 14 years old are able to perform quality chest compressions. Thus, classes from 14 to 18 years old were selected. Each session lasted 100 mins and was provided by nurses, with a ratio of one nurse to six students. Laerdal Medical's QCPR® software was used to measure and collect data of the quality of chest compressions. According to QCPR®, CPR is considered high quality if the final score is higher than 84%. A p-value of less than 0.05 was considered statistically significant. Results: A total of 734 young people were trained, with a median score of 97% (89–99) on the final QCPR® score. All students were able to provide chest compressions with the appropriate depth (>5 cm), 82% (44–96) provided chest compress at the recommended rate (100–120 bpm), and median compression rate of 112 (106–119) cpm. Male students performed deeper and more successful compressions but with less recoil than female students. Conclusions: School-age children aged 14–18 years old were able to learn and perform high-quality BLS maneuvers. We consider these results can have relevant public-health implications.
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Introduction: Due to the high morbidity and mortality associated with COVID-19, clinical risk stratification tools have been developed for these patients, in order to predict progression to severe disease and/or mortality. In this study, the Quick COVID19 Severity Index Scale was compared with News2 as a predictor of mortality in adults with SARS-CoV-2 infection, specifying its diagnostic validity. Materials and methods: Observational, retrospective, analytical study of diagnostic tests. 255 patients were included, over 18 years of age, hospitalized in Hospital II - I Moyobamba with a diagnosis of COVID-19, with an oxygen requirement of ≤6 Lt/min, from January to December 2020. Results: 13% (32/255) of the patients died, with a predominance of males 62.5% (20/32), with an average age of 50 years, the bioclinical parameters evaluated did not show a statistically significant association, except for the blood glucose p: 0.01. Regarding the scales, considering a high score ≥7 points;Quick Covid-19 Severity Index has no significant association as a predictor of mortality, with a sensitivity and specificity of 43.75% and 63.23%, respectively;while NEWS-2 showed sensitivity of 87.50%;specificity of 50.67%;AUC 0.768 (CI: 0.684-0.853, p 0.00);that is, there is more than 75% probability of prognosis. Conclusion: the NEWS-2 scale statistically showed to be a good tool for the prognosis of mortality in patients infected with SARS - CoV - 2, it is suggested to carry out studies that confirm the findings of this research and assess its applicability and reproducibility in other populations. © 2022 Instituto de Altos Estudios de Salud Publica. All rights reserved.