Single cell sequencing reveals cellular landscape alterations in the airway mucosa of patients with pulmonary long COVID (preprint)

To elucidate the important cellular and molecular drivers of pulmonary long COVID, we generated a single-cell transcriptomic map of the airway mucosa using bronchial brushings from patients with long COVID who reported persistent pulmonary symptoms. Adults with and without long COVID were recruited from the general community in greater Vancouver, Canada. The cohort was divided into those with pulmonary long COVID (PLC), which was defined as persons with new or worsening respiratory symptoms following at least one year from their initial acute SARS-CoV-2 infection (N=9); and control subjects defined as SARS-CoV-2 infected persons whose acute respiratory symptoms had fully resolved or individuals who had not experienced acute COVID-19 (N=9). These participants underwent bronchoscopy from which a single cell suspension was created from bronchial brush samples and then sequenced. A total of 56,906 cells were recovered for the downstream analysis, with 34,840 cells belonging to the PLC group. A dimensionality reduction plot shows a unique cluster of neutrophils in the PLC group (p<.05). Ingenuity Pathway Analysis revealed that neutrophil degranulation pathway was enriched across epithelial cells. Differential gene expression analysis between the PLC and control groups demonstrated upregulation of mucin genes in secretory cell clusters. A single-cell transcriptomic landscape of the small airways shows that the PLC airways harbors a dominant neutrophil cluster and an upregulation in the neutrophil-associated activation signature with increased expression of MUC genes in the secretory cells. Together, they suggest that pulmonary symptoms of long COVID may be driven by chronic small airway inflammation.

The underlying molecular mechanisms and biomarkers between periodontitis and COVID- 19 (preprint)

Objective Emerging evidence shows the clinical consequences of patient with COVID-19 and periodontitis are not promising, and periodontitis is a risk factor. Periodontitis and COVID-19 probably have a relationship. Hence, this study aimed to identify the common molecular mechanism that may help to devise potential therapeutic strategies in the future.Material and methods We analyzed two RNA-seq datasets for differential expressed genes, enrichment of biological processes, transcription factors (TFs) and deconvolution-based immune cell types in periodontitis, COVID-19 and healthy controls. Relationships between TFs and mRNA were established by Pearson correlation analysis, and the common TFs-mRNA regulatory network and nine co-upregulated TFs of the two diseases was obtained. The RT-PCR detected the TFs.Results A total of 1616 and 10201 DEGs from periodontitis and COVID-19 are found. Moreover, nine shared TFs and common biological processes associated with lymphocyte activation involved in immune response were identified across periodontitis and COVID-19. The cell type enrichment revealed elevated plasma cells among two diseases. The RT-PCR further confirmed the nine TFs up-regulation in periodontitis.Conclusion The pathogenesis of periodontitis and COVID-19 is closely related to the expression of TFs and lymphocyte activation, which can provide potential targets for treatment.

A versatile integrated tube for rapid and visual SARS-CoV-2 detection

The coronavirus disease 2019 (COVID-19) caused by novel severe acute respiratory coronavirus 2 (SARS-CoV-2) has been rapidly spreading worldwide. Rapid and widespread testing is essential to promote early intervention and curb the ongoing COVID-19 pandemic. Current gold standard reverse transcription-polymerase chain reaction (RT-PCR) for detecting SARS-CoV-2 is restricted to professional laboratories and well-trained personnel, thus, limiting its widespread use in resource-limited conditions. To overcome these challenges, we developed a rapid and convenient assay using a versatile integrated tube for the rapid and visual detection of SARS-CoV-2. The reaction conditions of the method were optimized using SARS-CoV-2 RNA standards and the sensitivity and specificity were further determined. Finally, it was verified on clinical specimens. The assay was completed within 40 min, and the result was visible by the naked eye. The limits of detection (LODs) for the target ORF1ab and N genes were 50 copies/μl. No cross-reactivity was observed with the RNA standard samples of four respiratory viruses or clinical samples of common respiratory viral infections. Ninety SARS-CoV-2 positive and 30 SARS-CoV-2 negative patient specimens were analyzed. We compared these results to both prior and concurrent RT-PCR evaluations. As a result, the overall sensitivity and specificity for detection SARS-CoV-2 were 94.5 and 100.0%, respectively. Conclusion The integrated tube assay has the potential to provide a simple, specific, sensitive, one-pot, and single-step assay for SARS-CoV-2.

Characterisation of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein. (preprint)

Pig producers have suffered serious economic losses from porcine epidemic diarrhea virus (PEDV) infection. Development of PEDV antibodies for detection or treatment has become an important research direction.The S1/S2 junction (S1S2J) cleavage site of S protein of PEDV is one of the major determinants of coronavirus infection success. In this study, we specifically selected the S1S2J protein of PEDV-AJ1102 (a representative strain of the G2 type) as target protein to immunise mice and generated monoclonal antibodies (mAbs) using hybridoma technology. Three mAbs showed high-binding activity to the S1S2J protein were further analysed. To reveal the characterisation of mAbs, variable region genes of antibodies were studied by using DNA sequencing, which revealed differences in their CDR3 amino acid sequences. We then developed a new method to identify the isotypes of these three mAbs. Results showed that these three antibodies were of the IgM type. As for the functions of these three mAbs, indirect immunofluorescence assay confirmed their good binding ability to Vero E6 cells infected with PEDV-SP-C strain (G1 type). Epitope analysis showed linear epitopes for all three mAbs. These antibodies were also used to detect infected cells via flow cytometry analysis. In summary, we have prepared and examined three mAbs against PEDV-S1S2J. The focal antibodies can be employed as detection antibodies for diagnostic reagents and further developed for other applications. We also designed a novel technique for easy and cost-saving identification of isotypes of mouse mAbs. Our results lay a good foundation for the development of research for PEDV.

Airport terminal passenger forecast under the impact of COVID-19 outbreaks: A case study from China

Passengers significantly affect airport terminal energy consumption and indoor environmental quality. Accurate passenger forecasting provides important insights for airport terminals to optimize their operation and management. However, the COVID-19 pandemic has greatly increased the uncertainty in airport passenger since 2020. There are insufficient studies to investigate which pandemic-related variables should be considered in forecasting airport passenger trends under the impact of COVID-19 outbreaks. In this study, the interrelationship between COVID-19 pandemic trends and passenger traffic at a major airport terminal in China was analyzed on a day-by-day basis. During COVID-19 outbreaks, three stages of passenger change were identified and characterized, i.e., the decline stage, the stabilization stage, and the recovery stage. A typical "sudden drop and slow recovery” pattern of passenger traffic was identified. A LightGBM model including pandemic variables was developed to forecast short-term daily passenger traffic at the airport terminal. The SHapley Additive exPlanations (SHAP) values was used to quantify the contribution of input pandemic variables. Results indicated the inclusion of pandemic variables reduced the model error by 27.7% compared to a baseline model. The cumulative numbers of COVID-19 cases in previous weeks were found to be stronger predictors of future passenger traffic than daily COVID-19 cases in the most recent week. In addition, the impact of pandemic control policies and passengers' travel behavior was discussed. Our empirical findings provide important implications for airport terminal operations in response to the on-going COVID-19 pandemic.

Acquisition and Dissemination of Anti-Epidemic Medical Prescriptions During the Outbreaks in the Qing Dynasty of China

During the epidemics in the Qing dynasty, many medical professionals, nonprofessionals, and social organizations collected and sorted medical prescriptions related to infectious diseases. These people also compiled, published, disseminated, and consulted related medical formularies. The above historical event can be viewed as the construction and dissemination of medical knowledge. They edited and published medical formularies on infectious diseases with lower cost and in flexible ways by taking the initiative and giving full scope to creativity. Diverse anti-epidemic medical prescriptions from these medical formularies can be used for infectious diseases in the event of the outbreaks. However, the therapeutic effects of classical prescriptions and folk recipes cannot be regarded as the same. The wide circulation of anti-epidemic medical prescriptions and medical formularies was essentially a process of epidemic prevention resource allocation. Not only did it enable many nonprofessionals to participate in epidemic prevention and control, but it also enhanced awareness, knowledge, and capacity for epidemic prevention at the individual level. At the same time, due to the uneven quality and individual differences in the physical fitness and condition of the prescriptions and formularies, they had the capacity of causing inconveniences to the readers or patients.

Associations between social support and anxiety during the COVID-19 lockdown in young and middle-aged Israelis: A cross-sectional study.

BACKGROUND: This study examined the associations between social support and anxiety during the coronavirus disease 2019 (COVID-19) in an Israeli sample. AIM: To examine the associations between social support and anxiety during the COVID-19 in an Israeli sample. METHODS: Data for this cross-sectional study were retrieved from an online survey. Linear regression, logistic regression and restricted cubic spline models were conducted to test for associations between social support and anxiety. RESULTS: A total of 655 individuals took part in the present study. In the univariate linear regression model, there is a negative correlation between the Generalized Anxiety Disorder-7 score (GAD-7) and the Multidimensional Perceived Social Support Scale (MSPSS) score. For MSPSS score, the multivariable adjusted regression coefficient and 95% confidence interval (CI) of GAD-7 score were -0.779 (-1.063 to -0.496). In the univariate logistic regression model, there was a negative correlation between anxiety (GAD-7 ≥ 9) and MSPSS score, and there was still a negative correlation in multivariate logical regression analysis. The odds ratios and 95%CI were 0.709 (0.563-0.894). CONCLUSION: Social support was inversely correlated with anxiety during COVID-19 in an Israeli sample.

Management model of mass vaccination of COVID-19 vaccine

OBJECTIVE: To explore the role of systematic vaccination management model in mass vaccination of COVID-19 vaccine. METHODS: According to the mass vaccination plan formulated by the Day Clinic of the Fifth Medical Center of Chinese PLA General Hospital, the vaccination work was fully deployed before vaccination;and the temporary specification of mass vaccination sites were set up according to the requirements of the temporary vaccination site for the new coronavirus vaccine in Beijing;the vaccine management systems was strictly implemented, the doctor group, nursing group and guarantee group performed the responsibilities of their own to ensure smooth and orderly vaccination work. RESULTS: The medical center has participated in more than 30 mass vaccinations of COVID-19, no vaccination error-related reactions and psychogenic reactions occurred at the vaccination site, and no adverse events that were caused by problems in the process link occurred. CONCLUSION: The detailed work plan for mass vaccination may facilitate the safe and orderly mass vaccination, and the vaccination work is unanimously recognized by the recipients.

Diagnostic Value of a Novel Eosinophil Cationic Protein-Myeloperoxidase Test Paper Before and After Treatment for Allergic Rhinitis.

Purpose: To develop and evaluate the effectiveness/clinical application of an eosinophil cationic protein-myeloperoxidase (ECP-MPO) test paper before and after treatment in patients with allergic rhinitis (AR). Patients and Methods: We included 40 controls and 106 AR patients who were enrolled in the Allergy Clinic of Renmin Hospital of Wuhan University. Total IgE, specific IgE and skin prick test (SPT) were detected in all participants. AR patients were treated with oral cetirizine hydrochloride for 14 days. The ECP-MPO test paper results, nasal secretion smear and eosinophil counts, rhinoconjunctivitis total nasal symptom score (TNSS), quality of life questionnaire (RQLQ), visual analogue scale (VAS), serum Th1/Th2/Th17 cytokine, and chemokine data were collected pre- and post-treatment. ECP concentrations in nasal secretions were assessed by ELISA. Pearson correlation test and Kappa consistency test were used for statistical analysis. Results: The post-treatment colour grade of the ECP-MPO test paper was lower in AR patients than the pre-treatment grade. The chromogenic grade correlated positively with the ECP concentration and the eosinophil count in nasal secretions both before and after treatment. Positive ECP-MPO test paper results were consistent with positive SPT, Der p-IgE and Der f-IgE result (Kappa values, 0.843, 0.810, 0.795, respectively). The pre- and post-treatment chromogenic grades correlated positively with the TNSS (r1=0·691; r2=0·539), RQLQ (r1=0·783; r2=0·625), and VAS (r1=0·703; r2=0·682) scores in AR patients. Conclusion: The ECP-MPO test paper can effectively detect ECP in nasal secretions, and its results are consistent with those from the SPT, Der p-IgE and Der f-IgE result. Its chromogenic grade can reflect the symptom severity and specific cytokine and chemokine levels in AR patients.

A Closed-loop Approach to Fight Coronavirus: Early Detection and Subsequent Treatment (preprint)

The recent COVID-19 pandemic has caused tremendous damage to social economy and people’s health. Some major issues fighting COVID-19 include early and accurate diagnosis and the shortage of ventilator machines for critical patients. In this work, we propose a closed-loop solution to deal with COVID-19: portable biosensing and wearable photoacoustic imaging for early and accurate diagnosis of infection and magnetically neuromodulation or minimally invasive electrical stimulation to replace the traditional ventilation. Proposed technique can guarantee ubiquitous and onsite detection, and electrical hypoglossal stimulator can be more effective in helping severe patients as well as reducing complications caused by ventilators.

Teaching Reform to the Biology Major During the COVID-19 Pandemic: A Study of the Method of Teaching Industrial Innovation and Entrepreneurial Talents.

The biology major has developed rapidly in recent years. Biology is a science that penetrates every aspect of human life and is one of the core majors in most agricultural colleges and universities. However, many teachers lack practical experience in the subject. To overcome this problem, in recent years, we have been trying to introduce new reforms into our teaching. This article provides some insight into the way that biology majors have been reformed, which will help educators in agricultural colleges and universities. At present, teachers implement the "Industrial Innovation and Entrepreneurship Talent Cultivation" (IIETC) model, but it is not clear whether this helps biology majors to master the course and improve their practical skills. In this study, the IIETC model is outlined, and the academic achievement and satisfaction of students taught under the IIETC model are assessed. A T-test is used to examine potential differences between IIETC and traditional teaching models. In-depth interviews and questionnaires were given to two groups of students who followed different teaching models as part of an exploratory study. The aim was to explore how effective IIETC is at helping biology majors master the course and improve students' wellbeing. Our results show that compared with traditional teaching methods, the IIETC model has a significant positive impact on the academic performance and happiness of biology students. Students trained under the IIETC model were more active and scored more highly in their final exams. They were more likely to feel that they had achieved success and happiness through the course (P = 0.03). The outcomes of this research reveal a novel teaching reform that improved students' enthusiasm for innovation and entrepreneurship during the ongoing COVID-19 pandemic. The effects are very encouraging and deserve further exploration and expansion in future work.

Lung spatial profiling reveals a T cell signature in COPD patients with fatal SARS-CoV-2 infection (preprint)

Rationale People with pre-existing lung diseases like chronic obstructive pulmonary disease (COPD) are more likely to get very sick from SARS-CoV-2 disease 2019 (COVID-19), but an interrogation of the immune response to COVID-19 infection, spatial throughout the lung structure is lacking in patients with COPD. Objectives To profile the immune microenvironment of lung parenchyma, airways, and vessels of never- and ever-smokers with or without COPD, whom all died of COVID-19, using spatial transcriptomic and proteomic profiling. Findings The parenchyma, airways, and vessels of COPD patients, compared to control lungs had: 1) significant enrichment for lung resident CD45RO + memory T cells; 2) downregulation of genes associated with T cell antigen-priming and memory T cell differentiation; 3) higher expression of proteins associated with SARS-CoV-2 entry and major receptor ubiquitously across the ROIs and in particular the lung parenchyma, despite similar SARS-CoV-2 structural gene expression levels. Conclusions The lung parenchyma, airways, and vessels of COPD patients have increased T-lymphocytes with a blunted memory T cell response and a more invasive SARS-CoV-2 infection pattern, and may underlie the higher death toll observed with COVID-19.

Open Reading Frame-3a gene of the 2019 novel coronavirus inhibits the occurrence and development of colorectal cancer.

Intestinal microecology is composed of bacteria, fungi and viruses. As a part of intestinal microecology, viruses participate in the occurrence and development of colorectal cancer. The 2019-nCoV was detected in stool samples from patients during COVID-19, suggesting that the 2019-nCoV may be associated with intestinal microecology. However, the relationship of the 2019-nCoV and CRC is unclear. The aim of this study is to explore the role of Open Reading Frame-3a (ORF3a) of the 2019-nCoV in CRC. After the pCDH-CMV-MCS-EF1-Puro vector that provides high expression of ORF3a was transfected into the SW480 CRC cell line, immunofluorescence was used to determine the localization of ORF3a in SW480 cells. The proliferation, migration, invasion, apoptosis, and cell cycle progression of SW480 cells were measured using the Cell Counting Kit-8 (CCK-8), wound healing, Transwell assay, flow cytometry, the TUNEL assay, and propidium iodide single staining. The results showed that ORF3a inhibited the proliferation, invasion, and migration of SW480 cells and induced their apoptosis after 24, 48, 72 h. Meanwhile, ORF3a inhibited the cell cycle and blocked SW480 CRC cells in the G1 phase. In in vivo experiments, high ORF3a expression was associated with decreased tumor volume, tumor weight, relative tumor volume, and tumor activity. ORF3a inhibited the growth and induced apoptosis and necrosis of tumor tissues. Based on this, we demonstrated that ORF3a might play a role in CRC, providing a new direction for the prevention and treatment of CRC.

Vaccine strain affects seroconversion after influenza vaccination in COPD patients and healthy older people.

Though clinical guidelines recommend influenza vaccination for chronic obstructive pulmonary disease (COPD) patients and other high-risk populations, it is unclear whether current vaccination strategies induce optimal antibody responses. This study aimed to identify key variables associated with strain-specific antibody responses in COPD patients and healthy older people. 76 COPD and 72 healthy participants were recruited from two Australian centres and inoculated with influenza vaccine. Serum strain-specific antibody titres were measured pre- and post-inoculation. Seroconversion rate was the primary endpoint. Antibody responses varied between vaccine strains. The highest rates of seroconversion were seen with novel strains (36-55%), with lesser responses to strains included in the vaccine in more than one consecutive year (27-33%). Vaccine responses were similar in COPD patients and healthy participants. Vaccine strain, hypertension and latitude were independent predictors of seroconversion. Our findings reassure that influenza vaccination is equally immunogenic in COPD patients and healthy older people; however, there is room for improvement. There may be a need to personalise the yearly influenza vaccine, including consideration of pre-existing antibody titres, in order to target gaps in individual antibody repertoires and improve protection.

Whole transcriptome analysis of high and low IFN-α producers reveals differential response patterns following rhinovirus stimulation.

OBJECTIVES: Viral respiratory infections cause considerable morbidity and economic loss. While rhinoviruses (RV) typically cause little more than the common cold, they can produce severe infections and disease exacerbations in susceptible individuals, such as those with asthma. Variations in the regulation of key antiviral cytokines, particularly type I interferon (IFN-α and IFN-ß), may contribute to RV susceptibility. To understand this variability, we compared the transcriptomes of high and low type I IFN producers. METHODS: Blood mononuclear cells from 238 individuals with or without asthma were cultured in the presence or absence of RV. Those samples demonstrating high or low RV-stimulated IFN-α production (N = 75) underwent RNA-sequencing. RESULTS: Gene expression patterns were similar in samples from healthy participants and those with asthma. At baseline, the high IFN-α producer group showed higher expression of genes associated with plasmacytoid dendritic cells, the innate immune response and vitamin D activation, but lower expression of oxidative stress pathways than the low IFN-α producer group. After RV stimulation, the high IFN-α producer group showed higher expression of genes found in immune response biological pathways and lower expression of genes linked to developmental and catabolic processes when compared to the low IFN-α producer group. CONCLUSIONS: These differences suggest that the high IFN-α group has a higher level of immune system readiness, resulting in a more intense and perhaps more focussed pathogen-specific immune response. These results contribute to a better understanding of the variability in type I IFN production between individuals.

To explore the impact of COVID-19 on HIV/AIDS case report in Nanning

Objective: To explore the impact of COVID-19 on HIV/AIDS case report in Nanning, and provide reference for the AIDS exploration during the period for prevention and control of COVID-19 epidemic.

Change of Consumption Behaviours in the Pandemic of COVID-19: Examining Residents' Consumption Expenditure and Driving Determinants.

This study investigated changes of individuals' consumption behaviours during the COVID-19 pandemic and explored the driving determinants in consumption expenditure in Zhejiang China. Based on the 454 samples of survey data, which were collected in 2020 and 2021, it showed a reduction trend in consumption expenditure during the pandemic. Compared to the consumptions before the pandemic, money spent on housing, food, and beverage did not change too much. However, expenditures on wearing, recreation, and education reduced. Age, family size, and household income were significant to the expenditure changes. Online shopping became an important alternative way for residents during the pandemic and the trend is expected to continue even after the pandemic. Based on the findings, suggestions are summarized as two points. First, the young and single residents are the main group for recovering the consumption for wearing, recreation, education, and public transport. Meanwhile, to improve the satisfactions in online shopping, regulations should be issued by the government in improving the quality of goods and service.

Deep Optical Blood Analysis: COVID-19 Detection as a Case Study in Next Generation Blood Screening (preprint)

A wide variety of diseases are commonly diagnosed via the visual examination of cell morphology within a peripheral blood smear. For certain diseases, such as COVID-19, morphological impact across the multitude of blood cell types is still poorly understood. In this paper, we present a multiple instance learning-based approach to aggregate high-resolution morphological information across many blood cells and cell types to automatically diagnose disease at a per-patient level. We integrated image and diagnostic information from across 236 patients to demonstrate not only that there is a significant link between blood and a patients COVID-19 infection status, but also that novel machine learning approaches offer a powerful and scalable means to analyze peripheral blood smears. Our results both backup and enhance hematological findings relating blood cell morphology to COVID-19, and offer a high diagnostic efficacy; with a 79% accuracy and a ROC-AUC of 0.90.

Pilot Study On The Value of Echocardiography Combined With Lung Ultrasound In The Diagnosis And Treatment of COVID-19 Pneumonia (preprint)

Background: This study aimed to investigate the relationship between echocardiography results and lung ultrasound score (LUS) in coronavirus diseases 2019 (COVID-19) pneumonia patients and to evaluate the impact of their combined application in the diagnosis and treatment of COVID-19 pneumonia.Methods: Hospitalized COVID-19 pneumonia patients who underwent lung ultrasound and echocardiography daily were included in this study. Patients with tricuspid regurgitation within 3 days of admission were enrolled, and the correlation and differences between their pulmonary artery pressure (PAP) and LUS on days 3, 8, and 13 were compared. The inner diameter of the pulmonary artery root and the size of the atria and ventricles were also observed.Results: Pulmonary artery pressure within 3 days (on day 3, 8 and 13) of admission was positively correlated with LUS (r = 0.448, p = 0.003; r = 0.738, p = 0.000; r = 0.325, p = 0.036). On day 8 the values of both PAP and LUS were higher than their corresponding values on days 3 and 13 (p < 0.01). On day 8 the positive rate for increased PAP and LUS was 92.9% (39/42) and 90.5% (38/42), respectively, and the combined positive rate for these two was 97.6% (41/42). On day 8 the inner diameters of the right atrium, right ventricle, and pulmonary artery differed significantly from their corresponding values on days 3 and 13 (p < 0.05).Conclusions: PAP is positively correlated with LUS. The two should be combined for a more informative assessment of the status of recovery from COVID-19 pneumonia.

Multi-omics highlights ABO plasma protein as a causal risk factor for COVID-19 (preprint)

SARS-CoV-2 is responsible for the coronavirus disease 2019 (COVID-19) and the current health crisis. Despite intensive research efforts, the genes and pathways that contribute to COVID-19 remain poorly understood. We therefore used an integrative genomics (IG) approach to identify candidate genes responsible for COVID-19 and its severity. We used Bayesian colocalization (COLOC) and summary-based Mendelian randomization to combine gene expression quantitative trait loci (eQTLs) from the Lung eQTL (n=1,038) and eQTLGen (n=31,784) studies with published COVID-19 genome-wide association study (GWAS) data from the COVID-19 Host Genetics Initiative. Additionally, we used COLOC to integrate plasma protein quantitative trait loci (pQTL) from the INTERVAL study (n=3,301) with COVID-19-associated loci. Finally, we determined any causal associations between plasma proteins and COVID-19 using multi-variable two-sample Mendelian randomization (MR). We found that the expression of 20 genes in lung and 31 genes in blood was associated with COVID-19. Of these genes, only three (LZTFL1, SLC6A20 and ABO) had been previously linked with COVID-19 in GWAS. The novel loci included genes involved in interferon pathways (IL10RB, IFNAR2 and OAS1). Plasma ABO protein, which is associated with blood type in humans, demonstrated a significant causal relationship with COVID-19 in MR analysis; increased plasma levels were associated with an increased risk of having COVID-19 and risk of severe COVID-19. In summary, our study identified genes associated with COVID-19 that may be prioritized for future investigation. Importantly, this is the first study to demonstrate a causal association between plasma ABO protein and COVID-19.