Predictors of Mortality among Patients Hospitalized with COVID-19 during the First Wave in India: A Multisite Case-Control Study.

Severe acute respiratory syndrome coronavirus 2 disease (COVID-19) has caused more than 6 million deaths globally. Understanding predictors of mortality will help in prioritizing patient care and preventive approaches. This was a multicentric, unmatched, hospital-based case-control study conducted in nine teaching hospitals in India. Cases were microbiologically confirmed COVID-19 patients who died in the hospital during the period of study and controls were microbiologically confirmed COVID-19 patients who were discharged from the same hospital after recovery. Cases were recruited sequentially from March 2020 until December-March 2021. All information regarding cases and controls was extracted retrospectively from the medical records of patients by trained physicians. Univariable and multivariable logistic regression was done to assess the association between various predictor variables and deaths due to COVID-19. A total of 2,431 patients (1,137 cases and 1,294 controls) were included in the study. The mean age of patients was 52.8 years (SD: 16.5 years), and 32.1% were females. Breathlessness was the most common symptom at the time of admission (53.2%). Increasing age (adjusted odds ratio [aOR]: 46-59 years, 3.4 [95% CI: 1.5-7.7]; 60-74 years, 4.1 [95% CI: 1.7-9.5]; and ≥ 75 years, 11.0 [95% CI: 4.0-30.6]); preexisting diabetes mellitus (aOR: 1.9 [95% CI: 1.2-2.9]); malignancy (aOR: 3.1 [95% CI: 1.3-7.8]); pulmonary tuberculosis (aOR: 3.3 [95% CI: 1.2-8.8]); breathlessness at the time of admission (aOR: 2.2 [95% CI: 1.4-3.5]); high quick Sequential Organ Failure Assessment score at the time of admission (aOR: 5.6 [95% CI: 2.7-11.4]); and oxygen saturation < 94% at the time of admission (aOR: 2.5 [95% CI: 1.6-3.9]) were associated with mortality due to COVID-19. These results can be used to prioritize patients who are at increased risk of death and to rationalize therapy to reduce mortality due to COVID-19.

Comparative study between first and second wave of COVID-19 deaths in India - a single center study (preprint)

Background The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is continuously evolving and many mutant variants of the virus are circulating in the world. Recurrent waves of COVID-19 have caused enormous mortality all over the world. It is of utmost importance for a health expert to understand the demographic and clinical attributes between the first and second waves of COVID-19 induced deaths. Method This was a hospital record based comparative study of baseline demographic, clinical and laboratory parameters of the first and second wave of COVID-19 in a tertiary care hospital in Uttarakhand, India. The study included all deceased patients admitted to the hospital during the first and second wave of COVID-19, i.e., between March 2020 to January 2021 and between March 2021 to June 2021, respectively. Result The study showed that there were more casualties in the second wave compared to the first, 475 (19.8%) and 424 (24.1%) respectively. There was no significant difference in terms of age. A male preponderance of mortality was evident in both the waves. The median duration of hospital stay was 5 (3-10) days in the second wave, which is significantly different from the corresponding duration in first wave (pConclusion In both the first and second COVID-19 waves, older males (>45 years) with comorbidities like HTN and DM were most susceptible for COVID-19 related mortality. The study also demonstrated that most of the baseline demographic and clinical characteristics which are attributed to the mortality were more common during the second wave of COVID-19.

A preventive study on hydroxychloroquine prophylaxis against COVID-19 in health care workers at a tertiary care center in north India

Background: The menace of COVID-19 has put a huge burden on health care system predisposing health care workers engaged in management to COVID-19 infections. The nonavailability of effective drug against COVID-19 warrants extra cover as prophylactic therapy for health care workers, especially against transmission from asymptomatic patients. Hydroxychloroquine (HCQ) for prophylaxis of COVID-19 had been advocated by some researchers. Hence, in this project, evaluation of HCQ as preventive strategy for healthcare workers against COVID-19 infection was studied. Materials and Methods: HCQ was prescribed as a prophylactic therapy as per the advisory of National Task Force of Indian Council of Medical Research, India. The data regarding consumption profile, COVID-19 infection and adverse drug reaction profile of HCQ in healthcare workers was collected.

Descriptive epidemiology of COVID-19 deaths during the first wave of pandemic in India - a single center experience (preprint)

BackgroundWith the looming threat of recurrent waves of COVID-19 in the presence of mutated strains, its of paramount importance to understand the demographic and clinical attributes of COVID-19 related mortalities in each pandemic waves. This could help policy makers, public health experts, and clinicians to better plan preventive and management strategies to curb COVID-19 related mortality. MethodThis was a hospital record based, retrospective cross-sectional descriptive study, at a tertiary care hospital in Rishikesh, India. The study included all deceased patients between March 2020 and January 2021 (first wave) who tested positive for SARS-CoV-2 by RT-PCR and were hospitalized. The study was done to describe demography, clinical presentation, laboratory parameters, treatment given and associated complications of all COVID-19 deaths. ResultOut of 424 mortalities, 298 (70.38%) were males and 126 (29.62%) were females. Mean age of patients was 55.85 {+/-} 16.24 years, out of which 19.5 percent were less than 45 years old, 33.6 percent were 45 to 60 years old and 41.8% were more than 60 years old. Comorbidity in the form of type 2 diabetes mellitus was present in 41.4% [95% CI (41.4-51.1)], hypertension in 39.8% [95% CI (35.1-44.6)], and coronary artery disease in 15.2% [95% CI (11.8-18.8)]. At the time of presentation, shortness of breath was present in 73.6% [95% CI (69.1 -77.7)], fever in 64.92% [95% CI (60.1-69.4)], and cough in 46.1%, [95% CI (41.1-50.8)]. Deranged laboratory parameters were lymphopenia in 90.2% [95% CI (86.8-92.7)], transaminitis in 59.7% [95% CI (54.8-64.3)], and hypercreatinemia in 37.7% [95% CI (33.1-42.5)]. Complications manifested were acute respiratory distress syndrome in 78.3% [95% CI (74-82.1)] and shock in 54.7% [95% CI (49.8-59.5)]. Median time duration between onset of symptom and hospital admission was 5 days (IQR = 3 - 5 days) and median length of hospital stay was 9 days (IQR = 4 - 14 days). ConclusionDuring first pandemic wave, COVID-19 related mortality was 2.37 times higher among males, 2.14 times in age group >60 than <45 years. Most common associated comorbidities (>40%) were type 2 diabetes mellitus and hypertension. Most common associated symptoms (>60%) were shortness of breath and fever. Lymphopenia was seen in >90% cases while liver involvement in 60% and kidney in 38% cases. Median hospital stay was doubled the pre-hospital illness.

SARS-CoV-2 Vaccine-Induced Antibody Response and Reinfection in Persons with Past Natural Infection (preprint)

Several studies have shown that subjects with a history of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection had significantly higher antibody titers than previously uninfected vaccinees after vaccination with mRNA vaccine. Yet no information is available for other vaccines. In the current observational cohort study, 105 health care workers who had received Covishield an Adeno associated viral vector-based DNA vaccine were enrolled at Sarojini Nadu Medical College Agra, India. The study included 40 (23 men and 17 women) subjects with a previous history of SARS-CoV-2 infection and 65 participants (39 men and 26 women) who were not infected previously. Both the groups received the adenovirus vector vaccine ChAdOx1-S recombinant vaccines (Covishield, Astra Zeneca). The levels of SARS-CoV-2-anti-spike-IgG-antibodies titer were measured using Electrochemiluminescence immunoassay on Roche platform as arbitrary units per milliliter (AU/ml). After 28 days of the second dose, subjects with no previous SARSCoV-2 infection showed a significantly lower level of circulating anti-spike-IgG-antibody titers compared to previously infected participants. After the second dose, we also observed a significant increase in SARS-CoV-2 infection in subjects with no prior history of SARS-CoV-2 infection compared to subjects with a previous history of natural infection. The most important observation of the study is a low percentage of infection in previously infected subjects. The finding of the study also indicates the presence of robust humoral memory response in previously infected patients.

The epidemiological and trending pattern of ncovid-19 in the state of Rajasthan, India

Purpose: The present study was undertaken to investigate the behavioral distribution pattern and progression of coronavirus disease 2019 (COVID-19) across age and gender in the state of Rajasthan, India, inherently distinctive and native to localized part of the globe giving requisite information and paraphernalia to designate advisory board of the state to design and frame customized policy for demands of the state as per the trending pattern relative to age and sex distribution, profile of new infected cases, recovery rate, and case fatality rate.

UVA radiation could be a significant contributor to sunlight inactivation of SARS-CoV-2 (preprint)

Past experiments demonstrated SARS-CoV-2 inactivation by simulated sunlight; models have considered exclusively mechanisms involving UVB acting directly on RNA. However, UVA inactivation has been demonstrated for other enveloped RNA viruses, through indirect mechanisms involving the suspension medium. We propose a model combining UVB and UVA inactivation for SARS-CoV-2, which improves predictions by accounting for effects associated with the medium. UVA sensitivities deduced for SARS-CoV-2 are consistent with data for SARS-CoV-1 under UVA only. This analysis calls for experiments to separately assess effects of UVA and UVB in different media, and for including UVA in inactivation models. Key words: SARS-CoV-2, COVID-19, environmental persistence, sunlight, UVA, UVB, modeling, inactivation methods, photobiology

Analysis of SARS-CoV-2 genomes from across Africa reveals potentially clinically relevant mutations. (preprint)

SARS-CoV-2 is a betacoronavirus, the etiologic agent of the novel Coronavirus disease 2019 (COVID-19). In December 2019, an outbreak of COVID-19 began in Wuhan province of the Hubei district in China and rapidly spread across the globe. On March 11th, 2020, the World Health Organization officially designated COVID-19 as a pandemic. Across the continents and specifically in Africa, all index cases were travel related. Thus, it is crucial to compare COVID-19 genome sequences from the African continent with sequences from COVID-19 hotspots (including China, Brazil, Italy, United State of America and the United Kingdom). To identify if there are distinguishing mutations in the African SARS-CoV-2 genomes compared to genomes from other countries, including disease hotspots, we conducted in silico analyses and comparisons. Complete African SARS-CoV-2 genomes deposited in GISAID and NCBI databases as of June 2020 were downloaded and aligned with genomes from Wuhan, China and other SARS-CoV-2 hotspots. Using phylogenetic analysis and amino acid sequence alignments of the spike and replicase (NSP12) proteins, we searched for possible targets for vaccine coverage or potential therapeutic agents. Our results showed a similarity between the African SARS-CoV-2 genomes and genomes in countries including China, Brazil, France, the United Kingdom, Italy, France and the United States of America. This study shows for the first time, an in-depth analysis of the SARS-CoV-2 landscape across Africa and will potentially provide insights into specific mutations to relevant proteins in the SARS-CoV-2 genomes in African populations.

A prefusion SARS-CoV-2 spike RNA vaccine is highly immunogenic and prevents lung infection in non-human primates (preprint)

To contain the coronavirus disease 2019 (COVID-19) pandemic, a safe and effective vaccine against the new severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is urgently needed in quantities sufficient to immunise large populations. In this study, we report the design, preclinical development, immunogenicity and anti-viral protective effect in rhesus macaques of the BNT162b2 vaccine candidate. BNT162b2 contains an LNP-formulated nucleoside-modified mRNA that encodes the spike glycoprotein captured in its prefusion conformation. After expression of the BNT162b2 coding sequence in cells, approximately 20% of the spike molecules are in the one-RBD up, two-RBD down state. Immunisation of mice with a single dose of BNT162b2 induced dose level-dependent increases in pseudovirus neutralisation titers. Prime-boost vaccination of rhesus macaques elicited authentic SARS-CoV-2 neutralising geometric mean titers 10.2 to 18.0 times that of a SARS-CoV-2 convalescent human serum panel. BNT162b2 generated strong TH1 type CD4+ and IFNy+ CD8+ T-cell responses in mice and rhesus macaques. The BNT162b2 vaccine candidate fully protected the lungs of immunised rhesus macaques from infectious SARS-CoV-2 challenge. BNT162b2 is currently being evaluated in a global, pivotal Phase 2/3 trial (NCT04368728).

In vitro study of BromAc on SARS-CoV-2 spike and envelope proteins shows synergy and disintegration at modest concentrations (preprint)

Objectives: SARS-CoV-2 infection is the cause of a worldwide pandemic, currently with limited therapeutic options. It is characterised by being highly contagious and nasal mucosa appears to be the primary site with subsequent spread to the lungs and elsewhere. BromAc (Bromelain & Acetylcysteine) has been described to disrupt glycoproteins by the synchronous breakage of glycosidic linkages and disulphide bonds. The spike protein of SARS-CoV-2 is an attractive target as it is essential for binding to the ACE2 receptor in host cells and is formed of glycoprotein and disulphide bridges for stabilisation. Hence, we sought to determine whether BromAc has activity on the spike and envelope protein specific to SARS-CoV-2 virus. Design: Gel electrophoresis analysis was carried out on recombinant spike and envelope proteins that were treated with a range of concentrations of single agents and BromAc. For UV analysis of disulfide bonds reduction, both spike and envelope protein were treated with Acetylcysteine with the determination of loss of disulfide bonds. Results: Recombinant spike and envelope SARS-CoV-2 protein were fragmented by BromAc whilst single agents had minimal effect. Spike and envelope proteins disulphide bonds were reduced by Acetylcysteine. Conclusion: BromAc disintegrates the spike and envelope protein from SARS-CoV-2 and may render it non-infective. In vitro tests on live virus have been encouraging and clinical testing through nasal administration in patients with early SARS-CoV-2 infection is imminent.

Convalescent plasma in the management of moderate COVID-19 in India: An open-label parallel-arm phase II multicentre randomized controlled trial (PLACID Trial) (preprint)

ObjectivesConvalescent plasma (CP) as a passive source of neutralizing antibodies and immunomodulators is a century-old therapeutic option used for the management of viral diseases. We investigated its effectiveness for the treatment of COVID-19. DesignOpen-label, parallel-arm, phase II, multicentre, randomized controlled trial. SettingThirty-nine public and private hospitals across India. ParticipantsHospitalized, moderately ill confirmed COVID-19 patients (PaO2/FiO2: 200-300 or respiratory rate > 24/min and SpO2 [≤] 93% on room air). InterventionParticipants were randomized to either control (best standard of care (BSC)) or intervention (CP + BSC) arm. Two doses of 200 mL CP was transfused 24 hours apart in the intervention arm. Main Outcome MeasureComposite of progression to severe disease (PaO2/FiO2< 100) or all-cause mortality at 28 days post-enrolment. ResultsBetween 22nd April to 14th July 2020, 464 participants were enrolled; 235 and 229 in intervention and control arm, respectively. Composite primary outcome was achieved in 44 (18.7%) participants in the intervention arm and 41 (17.9%) in the control arm [aOR: 1.09; 95% CI: 0.67, 1.77]. Mortality was documented in 34 (13.6%) and 31 (14.6%) participants in intervention and control arm, respectively [aOR) 1.06 95% CI: -0.61 to 1.83]. InterpretationCP was not associated with reduction in mortality or progression to severe COVID-19. This trial has high generalizability and approximates real-life setting of CP therapy in settings with limited laboratory capacity. A priori measurement of neutralizing antibody titres in donors and participants may further clarify the role of CP in management of COVID-19. Trial registrationThe trial was registered with Clinical Trial Registry of India (CTRI); CTRI/2020/04/024775.

Antibody binding to SARS-CoV-2 S glycoprotein correlates with, but does not predict neutralization (preprint)

Convalescent plasma from SARS-CoV-2 infected individuals and monoclonal antibodies were shown to potently neutralize viral and pseudoviral particles carrying the S glycoprotein. However, a non-negligent proportion of plasma samples from infected individuals as well as S-specific monoclonal antibodies were reported to be non-neutralizing despite efficient interaction with the S glycoprotein in different biochemical assays using soluble recombinant forms of S or when expressed at the cell surface. How neutralization relates to binding of S glycoprotein in the context of viral particles remains to be established. Here we developed a pseudovirus capture assay (VCA) to measure the capacity of plasma samples or antibodies immobilized on ELISA plates to bind to membrane-bound S glycoproteins from SARS-CoV-2 expressed at the surface of lentiviral particles. By performing VCA and neutralization assays we observed a strong correlation between these two parameters. However, while we found that plasma samples unable to capture viral particles did not neutralize, capture did not guarantee neutralization, indicating that the capacity of antibodies to bind to the S glycoprotein at the surface of viral particles is required but not sufficient to mediate neutralization. Altogether, our results highlights the importance of better understanding the inactivation of S by plasma and neutralizing antibodies.

Interaction of human ACE2 to membrane-bound SARS-CoV-1 and SARS-CoV-2 S glycoproteins (preprint)

A novel severe acute respiratory (SARS)-like coronavirus (SARS-CoV-2) is responsible for the current global coronavirus disease 2019 (COVID-19) pandemic, infecting millions of people and causing hundreds of thousands of deaths. The viral entry of SARS-CoV-2 depends on an interaction between the receptor binding domain of its trimeric Spike glycoprotein and the human angiotensin converting enzyme 2 (ACE2) receptor. A better understanding of the Spike/ACE2 interaction is still required to design anti-SARS-CoV-2 therapeutics. Here, we investigated the degree of cooperativity of ACE2 within both the SARS-CoV-2 and the closely related SARS-CoV-1 membrane-bound S glycoproteins. We show that there exist differential inter-protomer conformational transitions between both Spike trimers. Interestingly, the SARS-CoV-2 spike exhibits a positive cooperativity for monomeric soluble ACE2 binding when compared to the SARS-CoV-1 spike, which might have more structural restrains. Our findings can be of importance in the development of therapeutics that block the Spike/ACE2 interaction.

Restricting rural-urban connect to combat infectious disease epidemic as India fights COVID-19.

With declaration of 2019 novel coronavirus disease (COVID-19) as a pandemic on 11 March 2020 by World Health Organization, India came to alert for its being at next potential risk. It reached alert Level 2, i.e. local transmission for virus spread in early March 2020 and soon thereafter alert Level 3, i.e. community transmission. With on-going rise in COVID-19 cases in country, Government of India (GoI) has been taking multiple intense measures in coordination with the state governments, such as urban lockdown, active airport screening, quarantining, aggressive calls for 'work from home', public awareness, and active case detection with contact tracing in most places. Feedback from other countries exhibits COVID-19 transmission levels to have shown within country variations. With two-third of Indian population living in rural areas, present editorial hypothesizes that if India enters Level 3, rural hinterland would also be at risk importation (at least Level 1). Hence, we have to call for stringent containment on rural-urban and inter-state fringes. This along with other on-going measures can result in flattening curve and also in staggering 'lockdowns', and thus, helping sustain national economy.

Relationship Between ABO Blood group Phenotypes and nCOVID-19 Susceptibility – A Retrospective Observational Study (preprint)

Background: Since the outbreak of coronavirus disease-19 research has been continued to explore multiple facets of the disease. The objective of the present study is to evaluate the relationship between blood group phenotypes and COVID-19 susceptibility.Methods: In this hospital based, retrospective observational study 132 COVID-19 patients were enrolled from SMS Medical Hospital in Jaipur, India after receiving approval from the institutional ethics committee. The ABO, Rh and Kell blood group phenotypes along with demographic data of the patients were recorded. The observed proportions of ‘A’ , ‘B’, ‘AB’, ‘O’, ‘Rh’, and ‘Kell’ blood groups in COVID-19 patients were compared against the expected proportions (the null hypothesis) of the general population using Pearson’s chi-squared test and partition analysis.Results: There were significant differences between observed and expected frequency for the ABO and Kell blood phenotypes. Further partition analysis of ABO phenotypes showed that the group ‘A’ phenotypes were more susceptible to COVID-19. The Kell negatives were also more susceptible. The blood groups ‘AB’, ‘B’, ‘O’, and ‘Rh’ showed no significant difference for susceptibility to COVID-19.Conclusion: The study shows a relationship between ABO, Rh, and Kell blood groups and COVID-19 susceptibility. The application of these relationships in clinics should be explored in future studies.

Evaluation of Remote Monitoring Device for Monitoring Vital Parameters against Reference Standard: A Diagnostic Validation Study for COVID-19 Preparedness.

CONTEXT: Vital parameters including blood oxygen level, respiratory rate, pulse rate, and body temperature are crucial for triaging patients to appropriate medical care. Advances in remote health monitoring system and wearable health devices have created a new horizon for delivery of efficient health care from a distance. MATERIALS AND METHODS: This diagnostic validation study included patients attending the outpatient department of the institute. The accuracy of device under study was compared against the gold standard patient monitoring systems used in intensive care units. STATISTICAL ANALYSIS: The statistical analysis involved computation of intraclass correlation coefficient. Bland-Altman graphs with limits of agreement were plotted to assess agreement between methods. P <0.05 was considered statistically significant. RESULTS: A total of 200 patients, including 152 males and 48 females in the age range of 2-80 years, formed the study group. A strong correlation (intraclass correlation coefficient; r > 0.9) was noted between the two devices for all the investigated parameters with significant P value (<0.01). Bland-Altman plot drawn for each vital parameter revealed observations in agreement from both the devices. CONCLUSION: The wearable device can be reliably used for remote health monitoring. Its regulated use can help mitigate the scarcity of hospital beds and reduce exposure to health-care workers and demand of personal protection equipment.