Major role of S-glycoprotein in providing immunogenicity and protective immunity in mRNA lipid nanoparticle vaccines based on SARS-CoV-2 structural proteins (preprint)

Recently we have developed an mRNA lipid nanoparticle (mRNA-LNP) platform providing efficient long-term expression of an encoded gene in vivo after both intramuscular and intravenous application. Based on this platform, we have generated mRNA-LNP coding SARS-CoV-2 structural proteins M, N, S from different virus variants and studied their immunogenicity separately or in combinations in vivo. As a result, all candidate vaccine compositions coding S and N proteins induced excellent anti-RBD and N titers of binding antibodies. T cell responses mainly represented specific CD4+ T cell lymphocyte producing IL-2 and TNF-. mRNA-LNP coding M protein did not show high immunogenicity. High neutralizing activity was detected in sera of mice vaccinated with mRNA-LNP coding S protein (alone or in combinations) against closely related strains but was not detectable or significantly lower against an evolutionarily distant variant. Our data showed that the addition of mRNAs encoding S and M antigens to the mRNA-N in the vaccine composition enhanced immunogenicity of mRNA-N inducing more robust immune response to the N protein. Based on our results, we suggested that the S protein plays a key role in enhancement of immune response to the N protein in the mRNA-LNP vaccine.

Prolonged stay and endothelial dysfunction in hospitalized patients with covid-19

Introduction: COVID-19 is not only a respiratory disease, produces a severe systemic and multi-organ response. This illness generates vascular disorders, leading the patient to endothelial dysfunction. It acutely and chronically affects the patient's evolution, prolonging the patient's stay and worsening life prognosis. Objective(s): To evaluate differences in endothelial dysfunction present in patients hospitalized for COVID-19 who had a hospital stay longer than 18 days compared to those who did not. Method(s): A prospective cohort study was conducted. Hospitalized patients with confirmed SARS-COV 2 andolder than 18 years were included. Subjects in whom endothelial function markers could not be processed wereexcluded. Endothelial dysfunction was evaluated using E-selectin, endothelin-1, glutathione-s-transferase, arginase, and MDAM. A prolonged hospital stay was established >=18 days. Result(s): A total of 165 patients were evaluated, the average age of the population was 57.18 +/- 13.37 years, 73.33% were men. Subjects with prolonged hospital stay were older (59.38 +/- 12.08 vs 51.15 +/- 14.96, p=0.004), a higher number of patients required intubation (87.60 % vs 75, p=0.049) and e-selectin (1 [0.79 - 1.32] vs 0.88 [0.68 -1.14], p=0.0323) compared to subjects without prolonged hospital stay. Conclusion(s): Hospitalized patients over 18 days showed elevated levels of E-selectin reflecting endothelial damage, affecting vascular homeostasis, added to this, a significant number of them were intubated, increasing the risk of mortality, as well as future cardiovascular complications.

Patient and caregiver satisfaction regarding telepractice versus in-person services at a university speech, language, and hearing clinic

Purpose: The purpose of the pilot study is to explore patient and caregiver (i.e., spouse, parent, grandparent, child, and grandchild) satisfaction regarding telepractice versus in-person services provided at a university speech, language, and hearing clinic. Methods: Survey data were collected from 70 respondents (24 received in-person services only, 13 received telepractice only, and 33 received both in-person services and telepractice) during the onset of the COVID-19 pandemic. Results: Results indicated that patients and caregivers rated overall in-person services significantly higher than the overall telepractice services. Patients' and caregivers' perceived technology skills were not significantly related to their satisfaction of telepractice. However, graduate clinicians' technology skills, professionalism, and the quality of materials used in sessions were significantly associated with patients' and caregivers' satisfaction of telepractice. Also, patients' and caregivers' interaction with graduate students and supervisors was significantly correlated to their satisfaction of telepractice. Conclusions: Areas of improvement for telepractice-based services were discussed. The significance of adopting telepractice clinical and technical skills in preservice speechlan¬guage pathology pedagogy was also highlighted. © 2022 The Korean Association of Speech-Language Pathologists.

Serum markers of endothelial dysfunction and invasive mechanical ventilation in COVID-19

Introduction: Sars-Cov-2 infection induces vascular endothelium damage at pulmonary and systemic level. Alterations on immunity response generate inappropriate endothelial activation through pro-inflammatory cytokines release, up-regulated expression of adhesion molecules, leukocyte migration, increased nitric oxide requirements and oxidative stress. Endothelial function is a key feature in the pathogenesis of COVID-19. Objective(s): To evaluate circulating markers associated with endothelial activation in hospitalized patients with COVID-19 and determine the difference between patients who required invasive mechanical ventilation (IMV) and those who did not. Method(s): Cross-sectional study. Subjects with a confirmed diagnosis of COVID-19 and >18 years old were included. Patients who did not require hospitalization were excluded. Serum markers of endothelial function were tested during hospitalization and protein adjustment was performed. Result(s): A total of 390 patients were studied, with an average age of 57+/-13 years old. Patients who required invasive mechanical ventilation had higher prevalence of diabetes (34.53% vs 11.54%;p=0.020), higher serum nitrite levels (0.028 mmol/L [0.094-0.647] vs 0.07 [0.03-0.24];p=0.003), nitrates (0.363mmol/L [0.100- 0.591] vs 0.130[0.003-0.374];p=0.004) and E-selectin (1.00 ng/mg [0.79-1.32] vs 0.84 [0.55-1.09];p=0.019) when compared to non-IMV patients. Higher levels of nitrites adjusted by proteins were associated with an increased risk for IMV (OR 5.59, CI 95 1.15-27.00, p=0.032). Conclusion(s): Patients with increased nitrites and E-selectin levels had worse endothelial dysfunction and a higher risk for IMV during hospitalization.

Similar pulmonary functional outcomes at 3 months in critical COVID-19 survivors of the first, second and third pandemic waves

Introduction: We don't known whether pharmacological treatments and adaptation of ventilatory support has changed the outcomes of intensive care unit (ICU) COVID-19 survivors during the consecutive waves (W) of the pandemics. Aims and objectives: To assess pulmonary functional outcomes, radiologic pattern and quality of life (QoL) in ICU COVID-19 survivors at 3 months, according to W of pandemics. Method(s): Patients admitted to ICU for COVID-19 acute respiratory distress syndrome (ARDS) in 2 university hospitals were prospectively included, and assessed 3 months post discharge by a chest CT Scan, pulmonary function test (PFT), 6-minute walking distance test (6MWDT), respiratory muscle strength (RMS), and Short Form 36 (SF-36) questionnaire. Result(s): 84 ARDS COVID -19 survivors were included. Hospital length of stay was shorter during W3 vs W1 (23.4 +/- 14.2 vs 34.7 +/- 20.8, p= 0.03). Less patients required mechanical ventilation (MV) during W2 vs W1 (33.3 % vs 63.9%, p=0.0038). Three months after discharge, PFT, 6MWDT and RMS were similar, regardless the W (p>0.05). QoL (SF-36) was worse for patients of W1 vs W3 (64.7+/- 16.3 vs 49.2 +/- 23.2, p= 0.0169). On multivariate analysis, MV was associated with decreased total lung capacity (TLC), forced expiratory volume in 1 sec (FEV1), diffusing capacity of CO (DLCO) and RMS (w1,2,3, p<0.05). Low SF-36 score was correlated with low 6MWDT (w3, p= 0.01). Glucocorticoids use was associated with better PFT and chest CT recovery (p=0.00001), and tocilizumab with higher TLC (w1, p=0.03). Remdesevir improved MV duration in w2 (p=0.008). Conclusions At 3 months, PFT remains similar in ICU survivors, regardless pandemic w.

Impact of Improper Storage of ChAdOx1-S (AstraZeneca) Vaccine on Its Efficacy and Safety.

Background: In May 2021, there was an incident regarding giving patients AstraZeneca vaccines stored improperly. They were stored at room temperature (21 degrees centigrade) for 18 h, 12 h longer than the producer recommends. Aim of the study: The paper aims to contribute to the body of knowledge concerning the efficacy and safety of the ChAdOx1-S (AstraZeneca) vaccine concerning the requirements for cold supply chain specification. Patients and methods: Improperly stored vaccines were given to 44 patients, and 39 of them decided to take part in the study. The Control group consisted of 56 people vaccinated on the same days by the same medical teams, using properly stored medicines. Results: The concentration of anti-S1 SARS-CoV-2 Spike protein IgG antibodies did not differ significantly between the groups. Examined group median 70 kU/L (20;100). Control group median 66 kU/L (32.75;100), p = 0.751. We did not observe any COVID-19 infections in either the control or examined group for half a year after the incident. People from each group reported that local and systemic adverse events occurred directly after the first and second doses. In the control group, one case of spontaneously subsiding face edema and joint pain was observed. There were no severe or fatal adverse events. There were no significant differences between the groups, besides the fatigue, after the second dose. Conclusion: AstraZeneca vaccine ChAdOx1-S stored at 21 degrees centigrade for 18 h before vaccination has the same safety profile (p < 0.05) and the same efficacy (p < 0.05) as the vaccines stored in conditions recommended by the producer.

Primary Teachers Difficulties Related to Compulsory Distance Education during COVID-19

Due to a lack of preparation, teachers were not emotionally or cognitively prepared to use new approaches. Teachers discovered that while teaching online, elementary school teachers face various challenges. Summarizing and categorizing the difficulties encountered by primary school teachers in compulsory distance education will add to the literature, ensuring the dissemination of online instruction technological tools at both the distance education and primary education levels. The study sought to categorize the difficulties encountered by primary teachers. In order to figure out the purpose of the study, the study data including primary teachers in the COVID-19 period was synthesized. The study was accepted as a meta-synthesis. The education database ERIC was searched using the keywords "Primary Teacher", "Distance Education", and "Online Education", and 79 studies were located as a result of the search. After eliminating ineligible studies, 23 of the 79 collected studies remained. As a result, students, parents, infrastructure, and teachers are all affected by the challenges. Cutting-edge technologies and a strong internet infrastructure are required for online education. Teachers are unable to maintain the minimum level of compulsory distance education due to limited internet connection and a lack of technological resources in many places. As a result of the strain, teachers felt inadequate. In many classrooms in poor countries, there is a lack of access to professional expertise and help for the use and integration of ICTs. COVID-19 has made this an issue for almost all schools, not just those in underdeveloped countries. The importance of online learning for elementary children and teachers has gotten considerably less attention. Teachers can take pedagogical diversity professional development training online and experience it as if they were students. The practical advantage of identifying the difficulties that teachers have faced is to aid teachers in overcoming such obstacles and to give the appropriate assistance. Successful distance education can only be achieved when teachers are supported in all areas of difficulty.

Spanish cardiac catheterization and coronary intervention registry. 31st official report of the Interventional Cardiology Association of the Spanish Society of Cardiology (1990-2021)

Introduction and objectives: The Interventional Cardiology Association of the Spanish Society of Cardiology (ACI-SEC) presents its annual activity report for 2021. Methods: All Spanish centers with catheterization laboratories were invited to participate. Data were collected online and were analyzed by an external company, together with the members of the ACI-SEC. Results: A total of 121 centers participated (83 public and 38 private). Compared to 2020, both diagnostic coronary angiograms and percutaneous coronary interventions (PCI) increased by 11,4% and 10,3%, respectively. The radial approach was the most used access (92,8%). Primary PCI also increased by 6.2% whereas rescue PCI (1,8%) and facilitated PCI (2,4%) were less frequently conducted. Transcatheter aortic valve implantation was one of the interventions with the most relevant increase. A total of 5720 transcatheter aortic valve implantation procedures were conducted with an increase of 34,9% compared to 2020 (120 per million in 2021 and 89,4 per million in 2020). Other structural interventions like transcatheter mitral or tricuspid repair, left atrial appendage occlusion and patent foramen oval closure also experienced a significant increase. Conclusions: The 2021 registry demonstrates a clear recovery of the activity both in coronary and structural interventions showing a relevant increase compared to 2020, the year of the COVID-19 pandemic.

The emerging association between boredom, COVID-19 anxiety, and aggressiveness in imposed prolonged social isolation

The WHO's declaration of COVID-19 as a global health threat has contributed to a range of precautionary measures, such as quarantine and social distancing, and in certain situations a 2-month lockdown, such as in the case of Romania. Humans are social organisms and our evolutionary psychosocial processes have developed to flourish in people's collective contexts. The social distancing measure needed to counter the pandemic is considered to elevate the feelings of social alienation for those who already suffer from loneliness. The objective of our research is to analyze the boredom mediation effect, which is a prevailing state of mind in home quarantine and social isolation, and the association between coronavirus anxiety and aggressiveness. We consider that boredom, in the light of this research, is a modern globalized phenomenon which has a cultural dimension, a specific mental state that is considered unpleasant, and a lack of stimulation that triggers behavioral, psychological, and social consequences. An online survey investigated 390 adult respondents from Romania, who were selected on the basis of convenience sampling. We used SPSS's version 20 Process Macro for multiple regression analysis of variance. The findings revealed that under the prolonged context of COVID-19 social isolation, boredom completely mediated the relationship between coronavirus anxiety and aggressiveness. Our current research brings evidence to broadening the scope of the social dimension of sustainability to aspects related to mediatization and culturalization. Results, discussions, and recommendations to improve the well-being of socially isolated individuals in order to better cope with the present global circumstances are presented. © 2022 Elsevier Inc. All rights reserved.

Perception of resident doctors on postgraduate education during the COVID-19 pandemic: a preliminary assessment

Introduction. The aim of this study was to assess the results of the impact of the first year of the COVID-19 (coronavirus disease 19) pandemic on the postgraduate hybrid educational process (virtual / e-learning and traditional) from a resident-centred perspective, in a group of general dentistry resident doctors. Materials and methods. A questionnaire with 32 items (Q1-Q32) was configured using Google Forms and filled online, anonymously. In the first section, demographic information was collected. The other sections collected data about: 1) technical support), 2) the impact of traffic restriction and self-isolation, 3) the impact of preclinical/clinical online training compared to the on-site one, 4) attendance of online theoretical courses, 5) the effect of preclinical training in laboratories, 6) the degree of online interactivity and 7) general satisfaction. Excepting demographics and Q1-Q3 items, all other parts used multi-choice responses based on a five-degree Likert scale. Results. Most resident doctors did not report any problem in connection to online courses, increased anxiety, troubled concentration, or decreased learning efficiency. Conclusions. Resident doctors considered part-time e-learning process as a viable approach capable of providing a qualitative education and that the reorganization of the preclinical activity allowed them to carry out more precise and efficient procedures in the clinic. © 2022, Amaltea Medical Publishing House. All rights reserved.

The novel liquid learning system and the online gap in academic performance

This research considers the innovative educational strategy known as the liquid learning system, which allows students attending classes either online or face-to-face. This system was implemented for the first time at a private European university in 2020 as a reaction to the Covid-19 pandemic. Emphasis is placed on the effect of the online choice on student academic performance. Using Instrumental Variables to control for self-selection bias, our findings show a significant gap in the form of lower grades for online students. Quantile regressions reveal that those in the lower tail of the grade distribution are the most adversely affected.

Transforming a public university hospital and its area of influence into a comprehensive resource in response to the COVID-19 pandemic.

INTRODUCTION AND OBJECTIVES: The outbreak of COVID-19 has overwhelmed healthcare systems all over the world. The aim of this article is to describe the process of transforming the Vall d'Hebron University Hospital, the second largest hospital in Spain, into a COVID-19 centre coordinating response to the pandemic in its reference area. MATERIALS AND METHODS: The study draws on the experience of the authors in transforming the hospital into a comprehensive resource in response to the COVID-19 pandemic. The strategy is based on four central strategies: early planning, coordination of all healthcare agents in its reference area, definition of clear leadership roles, and the organisation of care based on multidisciplinary teams with minimal recruitment of new staff. RESULTS: The transformation strategy enabled the hospital to cope with the surge in patients without exceeding its capacity. During the response phases, which amounted to a period of 57 days, 3106 patients consulted the ER and 2054 were admitted, 346 of whom were treated at the ICU. To accommodate the number of adult COVID-19 patients, adult ICU availability was progressive increased by 371%, and ordinary beds increased by 240. A total of 671 staff members went on sick leave after testing positive for COVID-19. CONCLUSION: The transformation experience of the hospital provides insight into how effectively adapt the structures and functioning of large hospitals. The relevance of territorial coordination during the pandemic is stressed as an effective strategy that contributed coping the pandemic.

Dual Inhibition of Vacuolar-ATPase and TMPRSS2 Is Required for Complete Blockade of SARS-CoV-2 Entry into Cells.

An essential step in the infection life cycle of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the proteolytic activation of the viral spike (S) protein, which enables membrane fusion and entry into the host cell. Two distinct classes of host proteases have been implicated in the S protein activation step: cell-surface serine proteases, such as the cell-surface transmembrane protease, serine 2 (TMPRSS2), and endosomal cathepsins, leading to entry through either the cell-surface route or the endosomal route, respectively. In cells expressing TMPRSS2, inhibiting endosomal proteases using nonspecific cathepsin inhibitors such as E64d or lysosomotropic compounds such as hydroxychloroquine fails to prevent viral entry, suggesting that the endosomal route of entry is unimportant; however, mechanism-based toxicities and poor efficacy of these compounds confound our understanding of the importance of the endosomal route of entry. Here, to identify better pharmacological agents to elucidate the role of the endosomal route of entry, we profiled a panel of molecules identified through a high-throughput screen that inhibit endosomal pH and/or maturation through different mechanisms. Among the three distinct classes of inhibitors, we found that inhibiting vacuolar-ATPase using the macrolide bafilomycin A1 was the only agent able to potently block viral entry without associated cellular toxicity. Using both pseudotyped and authentic virus, we showed that bafilomycin A1 inhibits SARS-CoV-2 infection both in the absence and presence of TMPRSS2. Moreover, synergy was observed upon combining bafilomycin A1 with Camostat, a TMPRSS2 inhibitor, in neutralizing SARS-CoV-2 entry into TMPRSS2-expressing cells. Overall, this study highlights the importance of the endosomal route of entry for SARS-CoV-2 and provides a rationale for the generation of successful intervention strategies against this virus that combine inhibitors of both entry pathways.

COVID-19 infection impact on cardiovascular and thrombotic events during acute phase and medium-term of the disease

Background: Coronavirus disease (COVID-19) was labelled a global pandemic in April 2020 by the World Health Organisation. By December of the same year the disease caused by SARS-COV-2, known as COVID-19 (Coronavirus disease 2019), had spread over 200 countries, infecting more than 70 million people, causing more that 1.5 million of deaths. Recent studies suggest SARS-CoV-2 infection may be related to cardiovascular and thrombotic events although the strength of association remains unclear. Aims: Evaluate the emergence of cardiovascular and thrombotic events (such as major acute cardiovascular events, ictus and other thrombosis) in the acute moment and in medium-term follow-up in COVID-19 patients. Methods: Single-Center, retrospective, observational study of cohorts based on all the inhabitants of the health area. Survival analysis of main outcomes (mortality, heart failure [HF], and major acute cardiovascular events-MACE -[a composite of cardiovascular mortality, myocardial infarction and stroke]) were adjusted by multivariate logistic regression. Results: Of the total population studied, 447,979 inhabitants, 1,030 (0.23%) were diagnosed with COVID-19 infection, of which 14,8% were smokers, 31,2% had high blood pressure (HTA), 12,8% had diabetes, 29,2% had dyslipidaemia, 2,7% had peripheral artery disease, 4,7% had ischemic heart disease, 3,3% had had a previous transient ischemic attack, 10% were in anti-aggregation treatment and 5,8% were in anticoagulation treatment at the time of diagnosis. Concerning the analytics middle values, the group treated with ACEI/ARAB had higher troponins and ferritin than the group without ACEI/ARAB treatment, whereas higher reactive C protein and D-dimer were found in this last group. The main results showed that COVID-19 infection had no effect regarding to cardiovascular and thrombotic disease on mortality (OR: 1.64, 95% CI 0.98 2.76, p=0.062), heart failure (OR: 0.98, 95% CI 0.53 1.79, p=0.942), thrombotic events (OR: 1.02, 95% CI 0.22 4.83, p=0.98) and major acute cardiovascular events (OR: 0.88, 95% CI 0.48 1.60, p=0.665). Conclusions: In conclusion, COVID-19 infection had no effect on the emergence of cardiovascular or thrombotic events taking into account the 6-month prognosis, defined as mortality, heart failure, or major acute cardiovascular events.

Fast track triage for COVID-19 based on a population study: The SODA score

Background: Healthcare systems are under prominent stress due to the COVID-19 pandemic. A fast and simple triage is mandatory to screen patients who will benefit from early hospitalization, from those that can be managed as outpatients. There is a lack of all-comers scores, and no score has been proposed for western-world population. Aims: To develop a fast-track risk score valid for every COVID-19 patient at diagnosis. Methods: Single-center, retrospective study based on all the inhabitants of a healthcare area. Logistic regression was used to identify simple and wide-available risk factors for adverse events (death, intensive care admission, invasive mechanical ventilation, bleeding >BARC3, acute renal injury, respiratory insufficiency, myocardial infarction, acute heart failure, pulmonary emboli, or stroke). Results: Of the total healthcare area population, 447.979 inhabitants, 965 patients (0.22%), were diagnosed with COVID-19. A total of 124 patients (12.85%) experienced adverse events. The novel SODA score (based on sex, peripheral O2 saturation, presence of diabetes, and age) demonstrated good accuracy for adverse events prediction (area under ROC curve 0.858, CI: 0.82-0.98). A cut-off value of <2 points identifies patients with low risk (positive predictive value [PPV] for absence of events: 98.9%) and a cut-off of >5 points, high-risk patients (PPV 58.8% for adverse events). Conclusions: This quick and easy score allows fast-track triage at the moment of diagnosis for COVID-19 using four simple variables: age, sex, SpO2, and diabetes. SODA score could improve preventive measures taken at diagnosis in high-risk patients and also relieve resources by identifying very low-risk patients.