A neonatal mouse model characterizes transmissibility of SARS-CoV-2 variants and reveals a role for ORF8.

Small animal models have been a challenge for the study of SARS-CoV-2 transmission, with most investigators using golden hamsters or ferrets. Mice have the advantages of low cost, wide availability, less regulatory and husbandry challenges, and the existence of a versatile reagent and genetic toolbox. However, adult mice do not robustly transmit SARS-CoV-2. Here we establish a model based on neonatal mice that allows for transmission of clinical SARS-CoV-2 isolates. We characterize tropism, respiratory tract replication and transmission of ancestral WA-1 compared to variants Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Delta (B.1.617.2), Omicron BA.1 and Omicron BQ.1.1. We identify inter-variant differences in timing and magnitude of infectious particle shedding from index mice, both of which shape transmission to contact mice. Furthermore, we characterize two recombinant SARS-CoV-2 lacking either the ORF6 or ORF8 host antagonists. The removal of ORF8 shifts viral replication towards the lower respiratory tract, resulting in significantly delayed and reduced transmission in our model. Our results demonstrate the potential of our neonatal mouse model to characterize viral and host determinants of SARS-CoV-2 transmission, while revealing a role for an accessory protein in this context.

Optimization of the molecular diagnosis of the acute hepatitis E virus infection.

To evaluate the diagnostic value of the combination of two broad-range PCR assays targeting two different and conserved regions of the viral genome for the diagnosis of acute Hepatitis E virus (HEV) infection. Patients with acute hepatitis were prospectively recruited. In all, HEV-IgM antibodies were tested together with evaluation of HEV viraemia by two PCR assays (ORF3 and ORF1). The number of individuals exhibiting negative IgM antibody results but carrying viral RNA was calculated by each PCR assay. Four-hundred and seventy individuals were included, of whom 145 (30.8%) were diagnosed as having acute HEV. Of them, 122 (84.1%) exhibited HEV-IgM antibodies, and 81 (55.8%) had detectable viral RNA for at least one PCR. Using the ORF3 molecular assay, 70 (48.3%) individuals were identified with HEV infection. When the ORF1 molecular assay was applied, 49 (33.8%) individuals were identified. The ORF3 assay detected viral RNA in 32 patients not detected by the ORF1 assay. In contrast, the ORF1 assay could amplify viral RNA in 11 patients who were not detected by the ORF3 assay. The parallel use of two broad-range PCR assays significantly increased the performance of the molecular diagnosis of HEV.

Can dogs serve as stress mediators to decrease salivary cortisol levels in a population of liberal arts college undergraduate students?

The steroid hormone cortisol can be used to measure physiological stress in humans. The hypothalamic-pituitary-adrenal (HPA) axis synthesizes cortisol, and a negative feedback cycle regulates cortisol depending on an individual's stress level and/or circadian rhythm. Chronic stress of college undergraduate students is associated with various adverse health effects, including anxiety and depression. Reports suggest that stress levels have risen dramatically in recent years, particularly among university students dealing with intense academic loads in addition to COVID-19 pandemic-related uncertainty. The increasing rate of mental illness on college campuses necessitates the study of mediators potentially capable of lowering stress, and thus cortisol levels. Research on mediation techniques and coping mechanisms have gained traction to address the concerning levels of stress, including the employment of human-animal interaction sessions on college campuses. In this study, human-canine interaction as a stress mediation strategy for undergraduate students was investigated. We measured salivary cortisol levels in 73 college undergraduate students during a 60-min interaction period with a dog to determine whether human-canine interactions are effective in lowering cortisol levels and potentially reducing chronic stress typical of undergraduate students. Our results indicate that a human-canine interaction for 60 min is an effective method for significantly reducing salivary cortisol and stress levels among undergraduate college students. These findings support the expansion of animal visitation programs on college campuses to help students manage stress.

Gut microbiome dysbiosis in antibiotic-treated COVID-19 patients is associated with microbial translocation and bacteremia.

Although microbial populations in the gut microbiome are associated with COVID-19 severity, a causal impact on patient health has not been established. Here we provide evidence that gut microbiome dysbiosis is associated with translocation of bacteria into the blood during COVID-19, causing life-threatening secondary infections. We first demonstrate SARS-CoV-2 infection induces gut microbiome dysbiosis in mice, which correlated with alterations to Paneth cells and goblet cells, and markers of barrier permeability. Samples collected from 96 COVID-19 patients at two different clinical sites also revealed substantial gut microbiome dysbiosis, including blooms of opportunistic pathogenic bacterial genera known to include antimicrobial-resistant species. Analysis of blood culture results testing for secondary microbial bloodstream infections with paired microbiome data indicates that bacteria may translocate from the gut into the systemic circulation of COVID-19 patients. These results are consistent with a direct role for gut microbiome dysbiosis in enabling dangerous secondary infections during COVID-19.

Evolution of the COVID-19 Pandemic after the Introduction of School Referral Nurses in the Province of Granada-A Descriptive Study.

The aim of this research is to describe the evolution of the pandemic in a school context, following the introduction of school nurses into the educational setting. BACKGROUND: The first wave of COVID-19 in Spain prevented social interaction by imposing lockdowns on the population. All non-essential activities, including face-to-face education, were interrupted, affecting the school-aged population during the second and third trimester of the 2019/2020 school year. Schools are places where prevention, identification and case management measures should be rapidly implemented. METHODS: This is a prospective and descriptive study using a quantitative method to study the school population of Granada and its province during the school year 2020/2021, from September 2020 to May 2021. The study participants were all schools under the jurisdiction of the Territorial Delegation of Education of Granada, whether public, private, state-subsidised or charter schools, including all educational stages. RESULTS: The confirmation rate in Granada city (11.2%), in contrast to the Andalusian average (6.9%), places Granada as the province with the highest incidence rate. The infection rates among teachers show the same confirmation rate as the general population of Granada (9%); however, among students this rate is lower (7%). There is a higher incidence of outbreaks taking place outside school and a lower incidence of outbreaks occurring within the school environment. Both partial and total outbreaks are more frequent in early childhood education. CONCLUSION: The early identification and management of reported suspected cases of COVID-19 in schools is proving effective in preventing infection in the school population, achieving good pandemic control in schools, and reducing the number of outbreaks and people affected. Schools have been confirmed to be safe. Establishing screening for asymptomatic schoolchildren could be a tool to improve control in schools.

The moderating role of sociodemographic and work-related variables in burnout and mental health levels of Mexican medical residents.

OBJECTIVE: To explore the moderating effects of sociodemographic and work-related variables on levels of burnout and mental health among medical residents. METHOD: A cross-sectional online survey was administered at the beginning of the second wave of COVID-19 at different public teaching hospitals where medical residents practiced in Mexico City. A total of 201 medical residents of different years completed the survey. RESULTS: Different univariate inferential analyses on the level of burnout and mental health indices showed significant differences between sex, marital status, previous reports of physical illness or psychological conditions, and residency ranking. However, the effect sizes of those differences were of low to medium size. A predictive path analysis revealed that the three stages of burnout (emotional exhaustion, depersonalization, and achievement dissatisfaction) negatively affect psychological wellbeing and positively affect psychological distress. Finally, even though sociodemographic variables showed some significant variation, the effect sizes were small and did not moderate the direct effect of burnout on mental health indices. CONCLUSIONS: Medical residents deling with every day medical situations, will be exposed to stressors that might increase the probability to experience emotional exhaustion. This would negatively affect levels of wellbeing and positively affect distress, despite their sociodemographic characteristics.

Persistent T-cell exhaustion in relation to prolonged pulmonary pathology and death after severe COVID-19: Results from two Norwegian cohort studies.

BACKGROUND: T-cell activation is associated with an adverse outcome in COVID-19, but whether T-cell activation and exhaustion relate to persistent respiratory dysfunction and death is unknown. OBJECTIVES: To investigate whether T-cell activation and exhaustion persist and are associated with prolonged respiratory dysfunction and death after hospitalization for COVID-19. METHODS: Plasma and serum from two Norwegian cohorts of hospitalized patients with COVID-19 (n = 414) were analyzed for soluble (s) markers of T-cell activation (sCD25) and exhaustion (sTim-3) during hospitalization and follow-up. RESULTS: Both markers were strongly associated with acute respiratory failure, but only sTim-3 was independently associated with 60-day mortality. Levels of sTim-3 remained elevated 3 and 12 months after hospitalization and were associated with pulmonary radiological pathology after 3 months. CONCLUSION: Our findings suggest prolonged T-cell exhaustion is an important immunological sequela, potentially related to long-term outcomes after severe COVID-19.

Viruses and Mycoplasma pneumoniae are the main etiological agents of community-acquired pneumonia in hospitalized pediatric patients in Spain.

OBJECTIVES: To describe the etiology of community-acquired pneumonia (CAP) in hospitalized children in Spain and analyze the predictors of the etiology. HYPOTHESIS: The different etiological groups of pediatric CAP are associated with different clinical, radiographic, and analytical data. DESIGN: Observational, multicenter, and prospective study. PATIENT SELECTION: This study included children aged 1 month to 17 years with CAP, who were hospitalized between April 2012 and May 2019. METHODS: An extensive microbiological workup was performed. The clinical, radiographic, and analytical parameters were analyzed for three etiological groups. RESULTS: Among the 495 children included, at least one causative pathogen was identified in 262 (52.9%): pathogenic viruses in 155/262 (59.2%); atypical bacteria (AB), mainly Mycoplasma pneumonia, in 84/262 (32.1%); and typical bacteria (TyB) in 40/262 (15.3%). Consolidation was observed in 89/138 (64.5%) patients with viral CAP, 74/84 (88.1%) with CAP caused by AB, and 40/40 (100%) with CAP caused by TyB. Para-pneumonic pleural effusion (PPE) was observed in 112/495 (22.6%) patients, of which 61/112 (54.5%) presented a likely causative pathogen: viruses in 12/61 (19.7%); AB in 23/61 (37.7%); and TyB in 26/61 (42.6%). Viral etiology was significantly frequent in young patients and in those with low oxygen saturation, wheezing, no consolidation, and high lymphocyte counts. CAP patients with AB as the etiological agent had a significantly longer and less serious course as compared to those with other causative pathogens. CONCLUSIONS: Viruses and M. pneumoniae are the main causes of pediatric CAP in Spain. Wheezing, young age, and no consolidation on radiographs are indicative of viral etiology. Viruses and AB can also cause PPE. Since only a few cases can be directly attributed to TyB, the indications for antibiotics must be carefully considered in each patient.

Oral saliva swab reverse transcription PCR for Covid-19 in the paediatric population.

OBJECTIVES: To evaluate the performance of oral saliva swab (OSS) reverse transcription PCR (RT-PCR) compared with RT-PCR and antigen rapid diagnostic test (Ag-RDT) on nasopharyngeal swabs (NPS) for SARS-CoV-2 in children. DESIGN: Cross-sectional multicentre diagnostic study. SETTING: Study nested in a prospective, observational cohort (EPICO-AEP) performed between February and March 2021 including 10 hospitals in Spain. PATIENTS: Children from 0 to 18 years with symptoms compatible with Covid-19 of ≤5 days of duration were included. Two NPS samples (Ag-RDT and RT-PCR) and one OSS sample for RT-PCR were collected. MAIN OUTCOME: Performance of Ag-RDT and RT-PCR on NPS and RT-PCR on OSS sample for SARS-CoV-2. RESULTS: 1174 children were included, aged 3.8 years (IQR 1.7-9.0); 73/1174 (6.2%) patients tested positive by at least one of the techniques. Sensitivity and specificity of OSS RT-PCR were 72.1% (95% CI 59.7 to 81.9) and 99.6% (95% CI 99 to 99.9), respectively, versus 61.8% (95% CI 49.1 to 73) and 99.9% (95% CI 99.4 to 100) for the Ag-RDT. Kappa index was 0.79 (95% CI 0.72 to 0.88) for OSS RT-PCR and 0.74 (95% CI 0.65 to 0.84) for Ag-RDT versus NPS RT-PCR. CONCLUSIONS: RT-PCR on the OSS sample is an accurate option for SARS-CoV-2 testing in children. A less intrusive technique for younger patients, who usually are tested frequently, might increase the number of patients tested.

Efficacy of short-course colchicine treatment in hospitalized patients with moderate to severe COVID-19 pneumonia and hyperinflammation: a randomized clinical trial.

Some patients with COVID-19 pneumonia develop an associated cytokine storm syndrome that aggravates the pulmonary disease. These patients may benefit of anti-inflammatory treatment. The role of colchicine in hospitalized patients with COVID-19 pneumonia and established hyperinflammation remains unexplored. In a prospective, randomized controlled, observer-blinded endpoint, investigator-initiated trial, 240 hospitalized patients with COVID-19 pneumonia and established hyperinflammation were randomly allocated to receive oral colchicine or not. The primary efficacy outcome measure was a composite of non-invasive mechanical ventilation (CPAP or BiPAP), admission to the intensive care unit, invasive mechanical ventilation requirement or death. The composite primary outcome occurred in 19.3% of the total study population. The composite primary outcome was similar in the two arms (17% in colchicine group vs. 20.8% in the control group; p = 0.533) and the same applied to each of its individual components. Most patients received steroids (98%) and heparin (99%), with similar doses in both groups. In this trial, including adult patients with COVID-19 pneumonia and associated hyperinflammation, no clinical benefit was observed with short-course colchicine treatment beyond standard care regarding the combined outcome measurement of CPAP/BiPAP use, ICU admission, invasive mechanical ventilation or death (Funded by the Community of Madrid, EudraCT Number: 2020-001841-38; 26/04/2020).

Persistence of SARS-CoV-2 total immunoglobulins in a series of convalescent plasma and blood donors.

BACKGROUND: The vast majority of COVID-19 cases both symptomatic and asymptomatic develop immunity after COVID-19 contagion. Whether lasting differences exist between infection and vaccination boosted immunity is yet to be known. The aim of this study was to determine how long total anti-SARS-CoV2 antibodies due to past infection persist in peripheral blood and whether sex, age or haematological features can influence their lasting. MATERIAL AND METHODS: A series of 2421 donations either of SARS-CoV-2 convalescent plasma or whole blood from 1107 repeat donors from January 2020 to March 2021 was analysed. An automated chemiluminescence immunoassay for total antibodies recognizing the nucleocapsid protein of SARS-CoV-2 in human serum and plasma was performed. Sex, age, blood group, blood cell counts and percentages and immunoglobulin concentrations were extracted from electronic recordings. Blood donation is allowed after a minimum of one-month post symptom's relapse. Donors were 69.7% males and their average age was 46. The 250 donors who had later donations after a positive one underwent further analysis. Both qualitative (positivity) and quantitative (rise or decline of optical density regarding consecutive donations) outcomes were evaluated. RESULTS AND DISCUSSION: In 97.6% of donors with follow-up, anti-SARS-CoV-2 protein N total antibodies remained positive at the end of a follow-up period of 12.4 weeks median time (1-46, SD = 9.65) after the first positive determination. The blood group was not related to antibody waning. Lower lymphocyte counts and higher neutrophils would help predict future waning or decay of antibodies. Most recovered donors maintain their total anti-SARS-CoV-2 N protein antibodies for at least 16 weeks (at least one month must have been awaited from infection resolution to blood donation). The 10 individuals that could be followed up longer than 40 weeks (approximately 44 weeks after symptom's relapse) were all still positive.

MED = MEDICAL SCIENCES

Lund University, Sweden and Nova Southeastern University ,Asthma, a chronic disorder of the lungs, is best thought of as an umbrella term for heterogenous pathologic processes including inflammation, remodeling and resulting reversible diminishment of air flow. Sarah Fontana, sgfontana@seu.edu MED-04 A conceptual study on the possible effects of Stromal Vascular Fraction stem cell treatment in patients with chronic migraines and tension headaches. In this study, magnetic resonance imaging (MRI) scans will be conducted on the brain of each patient suffering of migraines classified into four groups;Group one: no medication, Group two: prescribed chemical migraine medications, Group three: over the counter painkillers and anti-inflammatories, and Group four: revived SVF stem cells.

Safety of COVID-19 vaccination in patients with previous cerebral venous sinus thrombosis.

INTRODUCTION: Cerebral venous sinus thrombosis (CVST) after coronavirus (COVID-19) vaccination has been reported. There are no data about thrombosis risk in prior CVST patients. The objective of the study was to describe short-term serious adverse events to COVID-19 vaccines in patients with history of CVST. MATERIAL AND METHODS: We present an observational prospective study of patients with known CVST who received COVID-19 vaccination. Serious event rates within 30 days after second dose vaccination (except one dose for Janssen) were evaluated, including recurrences, hospital admission and death. RESULTS: The 62 vaccinated patients received: BNT162b2 (Pfizer-BioNTech) in 43 patients (69.4%), mRNA-1273 (Moderna) in 7 patients (11.3%), AZD1222 (ChAdOx1) in 7 patients (11.3%) and Ad26.COV2.S (Janssen) in 5 patients (8.1%). There were no thrombotic recurrences within 30 days of vaccination (95% confidence interval, 0.0-5.8). There was one death (1.6%), not attributable to the vaccine. CONCLUSIONS: COVID-19 vaccines are safe for previous CVST patients.

A Machine Learning Approach for the Differential Diagnosis between Sars-COV19 Infection and Influenza Viruses with Hematological Morphologic DATA (CELL MORPHOLOGIC DATA)

IntroductionHyperinflammatory response induced by the SARS-CoV19 (CV) coronavirus is the main cause of morbidity and mortality. Numerous studies have pointed-out the main role of monocyte activation. In addition neutrophils alterations appear to differ pathophysiologically from the changes that occur in Influenza Virus (IV) infection. Due to the overlap of symptoms between these two entities, the search of analytical markers that help in early diagnostic orientation is considered of crucial importance. Changes in cell function, phenotype, and morphology in circulating leukocytes can be translated into numerical data obtained from an automated analyzer. The objective of our study is to generate an Artificial Intelligence Model from conventional hematological blood count parameters which be able to discriminate between CV and IV infection, in a fast and efficient maner.MethodsThis is a retrospective single-center study, performed between January-April 2020. The patients (n = 816) were divided into two groups: Patients who came for suspected COVID and had a positive RT-PCR (n = 408) and patients with a diagnosis of influenza confirmed by RT-PCR (n = 408). The database was divided into two random subgroups (n = 654) to train the model and another (n = 162) to validate it.The first hemogram on admission to the Emergency Department of these patients was performed on a Beckman-Coulter® DXH-900 equipment. Total white blood cells, absolute neutrophils, absolute lymphocytes, absolute monocytes, monocyte distribution wide (MDW) and Cell Morphological Data (CMDs) based on the impedance, conductivity and light scattering of these leukocyte subpopulations have been used to construct the model.Five algorithms have been evaluated using the R studio Software and the Caret (Classification and Regression Training) package: Linear Discriminant Analysis (LDA), K-Nearest Neighbor (kNN), Neural Networks (NN), Support Vector Machines (SVM) and Recursive partitioning (Rpart).ResultsThe evaluation of the different models was based on the comparison of the efficacy obtained through a cross validation (10x). It was decided to choose the SVM model by presenting a median of the area under the ROC curve of 0,841. No data preprocessing was performed, and the parameters chosen for the model were: sigma = 0,014, C = 1 and Number of Support Vectors = 458.Parameters with greater importance (>80%) in the model, were CMDs based on Neutrophil Light Scattering (SDLNE, SDLAN, SDMNE and MNLNE).The analysis of results was performed using a confusion matrix, where the model predicts the diagnosis of each patient in the validation subgroup (Table 2). A ROC curve with an area of 0,892 was obtained, with a sensitivity and specificity of 80% and 85%, respectively (Fig 1).ConclusionsThe creation of prediction algorithms from hemogram parameters allow to discriminate between COVID 19 infection and influenza A and B with a high specificity and sensitivity in a fast way. This could be a great advance for the early diagnostic orientation and guide clinical decisions as soon as possible with the consequent clinical benefit.

SARS-CoV-2 Portrayed against HIV: Contrary Viral Strategies in Similar Disguise.

SARS-CoV-2 and HIV are zoonotic viruses that rapidly reached pandemic scale, causing global losses and fear. The COVID-19 and AIDS pandemics ignited massive efforts worldwide to develop antiviral strategies and characterize viral architectures, biological and immunological properties, and clinical outcomes. Although both viruses have a comparable appearance as enveloped viruses with positive-stranded RNA and envelope spikes mediating cellular entry, the entry process, downstream biological and immunological pathways, clinical outcomes, and disease courses are strikingly different. This review provides a systemic comparison of both viruses' structural and functional characteristics, delineating their distinct strategies for efficient spread.

Detection of SARS-CoV-2 RNA in serum is associated with increased mortality risk in hospitalized COVID-19 patients.

COVID-19 has overloaded national health services worldwide. Thus, early identification of patients at risk of poor outcomes is critical. Our objective was to analyse SARS-CoV-2 RNA detection in serum as a severity biomarker in COVID-19. Retrospective observational study including 193 patients admitted for COVID-19. Detection of SARS-CoV-2 RNA in serum (viremia) was performed with samples collected at 48-72 h of admission by two techniques from Roche and Thermo Fischer Scientific (TFS). Main outcome variables were mortality and need for ICU admission during hospitalization for COVID-19. Viremia was detected in 50-60% of patients depending on technique. The correlation of Ct in serum between both techniques was good (intraclass correlation coefficient: 0.612; p < 0.001). Patients with viremia were older (p = 0.006), had poorer baseline oxygenation (PaO2/FiO2; p < 0.001), more severe lymphopenia (p < 0.001) and higher LDH (p < 0.001), IL-6 (p = 0.021), C-reactive protein (CRP; p = 0.022) and procalcitonin (p = 0.002) serum levels. We defined "relevant viremia" when detection Ct was < 34 with Roche and < 31 for TFS. These thresholds had 95% sensitivity and 35% specificity. Relevant viremia predicted death during hospitalization (OR 9.2 [3.8-22.6] for Roche, OR 10.3 [3.6-29.3] for TFS; p < 0.001). Cox regression models, adjusted by age, sex and Charlson index, identified increased LDH serum levels and relevant viremia (HR = 9.87 [4.13-23.57] for TFS viremia and HR = 7.09 [3.3-14.82] for Roche viremia) as the best markers to predict mortality. Viremia assessment at admission is the most useful biomarker for predicting mortality in COVID-19 patients. Viremia is highly reproducible with two different techniques (TFS and Roche), has a good consistency with other severity biomarkers for COVID-19 and better predictive accuracy.