Evaluation of air quality and its variation due to COVID-19 lockdown in Hyderabad, Telangana, India

Air pollution in India is a serious health issue. A countrywide lockdown was imposed in India in response to the COVID-19 pandemic, firstly for three weeks starting from March 24 to April 14, 2020, and then extended until May 3, 2020. Because of the restrictions imposed, pollution levels in cities all over the country have dropped dramatically in just a few days, raised questions among scientists about lockdown as the most effective alternative approach for reducing air pollution. Hyderabad was chosen for this study because it is India's 5th largest city by area and 4th largest city by population, as well as major industrial centre in South-East Asia with strong air quality statistics. In light of the recent COVID-19 outbreak around the country, a detailed analysis based on air quality parameters from six distinct air quality monitoring sites in Hyderabad, Telangana, has been performed. For simple interpretation of air quality data, establishing a correlation between different pollutants, identifying sources of pollution, and determining the most significant parameters, different multivariate statistical approaches such as Cluster analysis (CA), Principle component analysis (PCA), correlation analysis, and multiple linear regression analysis (MLR) were used. The aim of this study is to evaluate the major air pollution sources in Hyderabad and to identify the most significant air pollutants based on their individual contributions to the Air Quality Index (AQI). Variation in air quality parameters collected for six air quality monitoring stations were represented using box or whisker plots. The data set has been grouped into four major clusters depending on the similarities in the air quality data. Major sources of air pollution in each cluster were identified using PCA. MLR analysis was used to create models for predicting AQI for each cluster based on concentrations of important air contaminants. The findings revealed that PM10 and PM2.5 play a significant role in determining AQI levels. © 2023 Author(s).

Attention to the Fads and Fashions in the Indian Stock Markets During COVID-19

Do people show fads and fashions in their attention searches? With the Google online search data during COVID-19, particularly from January to May 2020 for the socio-economic keywords, this study examines if online searches show short-run and long-run attention dynamics leading to fads and fashions in attention to the NSE Nifty and BSE Sensex indices. This study employs the methodology of cointegrating relationship with autoregressive distributed lag (ARDL) model and explains investors' attention search dynamics at the 'NSE Nifty Index' and 'BSE Sensex Index' caused by socio-economic attention searches. It also examines if the dynamics of attention coordination are parsimonious in nature and it explores the same with the generalized autoregressive conditional heteroskedastic (GARCH-X) model. With the ARDL models, this study finds robust and unbiased cointegrating impacts of socio-economic attention searches on the attention search for the NSE Nifty index but these are not the best linear unbiased and efficient (BLUE) ones, while the same on the BSE Sensex Index are BLUE. For the NSE Nifty index, the attention dynamics at the GARCH-X specification are BLUE while for the BSE Sensex index, the GARCH-X specification also has some additional information in terms of the ARCH effect only.

Effect of subprime crisis on financing decisions and capital structure of Indian firms: a pre- and post- crisis study using system GMM methodology

Purpose: The subprime crisis (SPC) (2007–2008) has severely affected the economies across the globe. The Indian economy was also troubled because the SPC led to a sharp reduction in foreign trade and investment, a rise in the exchange rate volatility and disproportionate foreign-currency reserves. The present paper analyzes the financing pattern of Indian listed companies during the SPC. This study aims to ascertain the impacts of the SPC-2008 on the long-term and short-term financing decisions of Indian listed companies using novel data set and appropriate robust methodology. Design/methodology/approach: The study uses fixed effect model autoregressive of order 1 (FEM AR (1)) and system generalised method of moments (GMM) methodology on a sample data of 1,032 Indian non-financial listed companies on the Bombay Stock Exchange (BSE) for the period 1999 to 2019 to analyze the financing pattern during the crisis. Findings: The study finds that the Indian firms opted for de-leveraging, shortening the maturity of debt and short-term borrowing. This significant decline in the leverage and maturity of debt indicates that the companies in India generally followed the "rat race” model of the financing mix in the crisis. After the crisis, the firms have re-leveraged and expanded the maturity of debt up to 90%. This considerable expansion in leverage and maturity implies that the Indian firms are exposed to the "rollover risk.” This re-leverage risk is asymmetrically distributed for manufacturing and services firms. Manufacturing firms are found to be more exposed to this risk. Furthermore, tangibility, free cash flows and the liquidity available within the firms are the compelling elements of the financing decision during the crisis. Research limitations/implications: The study has not included the private firms and unorganized sectors in India. Moreover, the study has not analyzed disasters such as the Asian liquidity crisis, the information technology (IT) bubble crisis, the euro bond crisis and coronavirus disease 2019 (COVID-19) pandemic. Practical implications: The study finds that Indian firms are exposed to higher risk during the financial crisis and this risk is further aggravated by the rollover risk. Therefore, investors and creditors should consider these additional risks in the financial decisions and take more precautions. The study suggests that the regulators should make necessary adjustments in lending policy, corporate restructuring and tax policy to deal with the menace of a financial crisis. Social implications: Indian firms should avoid following the rate race financing model. Originality/value: This study aims to ascertain the impacts of the SPC-2008 on the long-term and short-term financing decisions of Indian listed companies using novel data set and appropriate robust methodology. © 2023, Emerald Publishing Limited.

Cotrimoxazole in hospitalised patients with severe COVID-19 - interim results from the CoTroxCov study

Background: Severe COVID-19 has been attributed to a hyperimmune response mediated by cytokines. The mainstay of therapy remains largely supportive along with steroids. Co-trimoxazole in addition to having antimicrobial properties has immunomodulatory and anti-inflammatory properties and could potentially improve outcomes in patients with severe COVID-19 . Hypothesis: We hypothesised that Co-trimoxazole given to patients with severe COVID-19 could prevent progression to critical illness, mortality and reduce time to recovery. Method(s): We conducted an interim analysis in our single center open-label randomised control trial, in which hospitalised patients with severe COVID-19 requiring supplemental oxygen via non-rebreathe mask between 10 -15 Litres per minute and maintaining saturations between 92-96% were assigned in a 1:1 ratio to receive either oral Cotrimoxazole in addition to standard therapy or standard therapy alone. Result(s): 111 patients were recruited into the study, of which 56 patients received Co-trimoxazole and 55 received standard therapy alone. The mean age was 50 years in the Co-trimoxazole group versus 53 years in the standard therapy group (p=0.083). In-hospital mortality was 11% in the Co-trimoxazole group vs 29% in the standard therapy group (p=0.020). Mechanical ventilation was offered to 9% of the patients in the Co-trimoxazole group versus 13% of the patients in the standard therapy group. Time to recovery was 6 days in the Co-trimoxazole group versus 7 days in the standard therapy group (p=0.466). Conclusion(s): In this interim analysis oral Co-trimoxazole reduces mortality in patients with severe Covid-19. Further recruitment is underway.

Impact of COVID-19 on death anxiety in patients with anxiety spectrum disorders: A case-control study

Background: COVID-19 pandemic has exacerbated symptoms in existing diagnosed cases of anxiety. Its impact on people with mental illness is expected to be significant, leading to an increase in relapse rates and expectedly inducing or exacerbating death anxiety. Aims & Objectives: This study aimed to assess the impact of Covid 19 fear on general anxiety and wellbeing and death anxiety and compare people with anxiety spectrum disorders on perceived fear, autonomic anxiety symptoms, and death anxiety with those without these disorders. Materials & Methods: We recruited 36 participants with anxiety spectrum disorder (generalized anxiety disorder (n=6), obsessive-compulsive disorder (n=17), panic disorder (n=5) and mixed anxiety and depression (n=8) and 36 individuals free from any psychiatric illness. Tools used include General Health Questionnaire, Fear of COVID-19 Scale, Perceived Stress Scale, Beck Anxiety Inventory and Multidimensional Fear of Death Scale. Results: A statistically significant correlation was noted between many variables in both the study groups. The regression analysis showed a significant difference in general well-being and anxiety & death anxiety in the clinical group. Study outcomes indicated that the current pandemic has triggered significant fear and anxiety in anxiety patients and among healthy controls and has triggered significant death anxiety in the clinical and healthy control group. Conclusion: The study showed that fear of covid-19 is a predictive factor for stress, death anxiety, and general well-being. These findings may be helpful to plan preventive measures, tailored intervention focusing on death anxiety and relapse plans effectively. © 2022 by the Author(s).

Role of Chest Radiography as the Imaging Tool in COVID Pandemic Era

Background: The present radiological COVID literature is mainly confined to the CT findings. Using High Resolution Computed tomography (HRCT) as a regular 1st line investigation put a large burden on radiology department and constitute a huge challenge for the infection control in CT suite. Materials and Methods: A prospective study of 700 consecutive COVID positive cases who underwent Chest Xray (CXR) and HRCT thorax were included in the study. Many of these CXR were repeated and followed up over a duration of time to see the progression of disease. Results: 392/700 (56%) were found to be negative for radiological thoracic involvement. 147/700 (21%) COVID positive patients showed lung consolidations, 115/700 (16.5%) presented with GGO, 40/700 (5.7%) with nodules and 42/700 (6%) with reticular–nodular opacities. 150/700 patients (21.4 %) had mild findings with total RALE severity score of 1-2. More extensive involvement was seen in 104/700 (14.8 %) and 43/700 (6.2%) patients, who had severity scores of 3-4 and 5-6 respectively. 11/700 patients had a severity score of >6 on their baseline CXR. Those with severity score of 5 or more than 5 (54/700, 7.7%) required aggressive treatment with mean duration of stay of 14 days, many of them died also (23/54, 42.5%). Conclusion: In cases of high clinical suspicion for COVID-19, a positive CXR may obviate the need for CT. Additionally, CXR utilization for early disease detection and followup may also play a vital role in areas around the world with limited access to CT and RT-PCR test.

A study to evaluate the effect of the COVID-19 pandemic on the mental health of junior doctors of a Government Medical College in West Bengal

Background: The COVID-19 pandemic had posed huge challenges to junior doctors as they had to work as frontline health workers during this emergency. Objectives: To evaluate the mental health status of junior doctors during the present pandemic using Depression, Anxiety, Stress Scale-21 (DASS-21) and impact of even scale-revised (IES-R) scale scores. Methods and Participants: This longitudinal study was conducted over a time period of 8 months after taking Institutional Ethical Clearance and informed consent of 220 participants. This online survey was conducted using Google Forms and DASS-21 and IES-R scores were evaluated twice in an interval of 6 months. Results: Depression scores, anxiety scores, and stress scores were significantly increased on the second assessment. Gradation of depression, anxiety, and stress in month 1 among participants were as follows: Depression scores (normal;mild;moderate;severe;extremely severe): 82;40;52;33;13. Anxiety scores (normal;mild;moderate;severe;extremely severe): 113;21;55;16;15. Stress scores (normal;mild;moderate;severe;extremely severe): 137;33;41;8;1. ISE-R scores: <24 = 98;24-32 = 39;33-36 = 15;≥37 = 68. Gradation of depression, anxiety, and stress in month 6 were the following: Depression scores (normal;mild;moderate;severe;extremely severe): 50;19;63;26;62. Anxiety scores (normal;mild;moderate;severe;extremely severe): 34;5;67;25;89. Stress scores (normal;mild;moderate;severe;extremely severe): 61;12;42;38;67. ISE-R Scores: <24 = 73;24-32 = 34;33-36 = 11;≥37 = 102. Conclusions: Junior doctors working during the COVID-19 pandemic developed increased levels of stress, anxiety, and depression with the progression of the pandemic. They were the major frontline health workers to combat COVID-19 which may be the cause of their decreased psychological resilience resulting in mental health issues. Psychological counseling sessions can be used among these young doctors to build up their mental resilience. © 2022 by the Author(s).

Analysis of Neuroimaging in COVID-19 Patients - A Correlation Between Lung Severity and Brain Findings

Background: Corona virus emerged in China in December 2019 and quickly spread over the world, causing a pandemic. The probable link between the occurrence of neurological abnormalities and the CT severity score (CTSS) in COVID-19 participants is less understood. The purpose of this study was to look at the neurological symptoms of COVID-19 on CT head and determine whether there was a link between thorax and brain imaging abnormalities in COVID-19 patients. Method(s): Total 135 Hospitalized COVID positive patients with acute neurological symptoms underwent both CT head and CT thorax during their hospital stay were included in the study. All the patients with neuroimaging were divided into 2 groups: first being patients with acute neuroimaging findings and the second being the patients with chronic/normal neuroimaging findings. Result(s): The most common CT head imaging findings in these individuals were acute ischemic infarcts in 54 (40%) and acute intracranial haemorrhage in 8 (6%). When compared to individuals with normal/chronic neurological results, a greater mean chest CTSS was found in patients with acute abnormalities on CT head (14.1 [SD-3.2] versus 6.5 [SD-3.3]). However, no statistical correlation could be shown between a greater CTSS and the occurrence of acute neurological disorders. Conclusion(s): There was no link between a greater CTSS and the occurrence of neurological disorders on CT scans. As a result, increased lung involvement severity may not be a good predictor of brain involvement in COVID patients. Copyright © 2022 Ubiquity Press. All rights reserved.

Role of Chest Radiography as the Imaging Tool in COVID Pandemic Era

Background: The present radiological COVID literature is mainly confined to the CT findings. Using High Resolution Computed tomography (HRCT) as a regular 1st line investigation put a large burden on radiology department and constitute a huge challenge for the infection control in CT suite. Material(s) and Method(s): A prospective study of 700 consecutive COVID positive cases who underwent Chest Xray (CXR) and HRCT thorax were included in the study. Many of these CXR were repeated and followed up over a duration of time to see the progression of disease. Result(s): 392/700 (56%) were found to be negative for radiological thoracic involvement. 147/700 (21%) COVID positive patients showed lung consolidations, 115/700 (16.5%) presented with GGO, 40/700 (5.7%) with nodules and 42/700 (6%) with reticular-nodular opacities. 150/700 patients (21.4 %) had mild findings with total RALE severity score of 1-2. More extensive involvement was seen in 104/700 (14.8 %) and 43/700 (6.2%) patients, who had severity scores of 3-4 and 5-6 respectively. 11/700 patients had a severity score of >6 on their baseline CXR. Those with severity score of 5 or more than 5 (54/700, 7.7%) required aggressive treatment with mean duration of stay of 14 days, many of them died also (23/54, 42.5%). Conclusion(s): In cases of high clinical suspicion for COVID-19, a positive CXR may obviate the need for CT. Additionally, CXR utilization for early disease detection and followup may also play a vital role in areas around the world with limited access to CT and RT-PCR test. Copyright © 2022, Dr Yashwant Research Labs Pvt Ltd. All rights reserved.

Safety and Immunogenicity Outcomes of an Inactivated Viral Vaccine against SARS-CoV-2 (Covaxin)

Introduction: Bharat Biotech International Ltd in partnership with National Institute of Virology (NIV), has developed an indigenous whole virion inactivated Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) viral vaccine BBV-152 (Covaxin), formulated with Toll Like Receptors 7/8 agonist Imidazoquinoline (IMDG) molecule adsorbed to alum (Algel). Variety of factors other than environmental ones can affect vaccines efficiency outside the strict setting of clinical trials, like how the vaccine is stored or transported, and even how patients are vaccinated. In addition, the intrinsic capacity of the recipient to respond to a vaccine which is determined by sex, genetic factors, age, psychological stress, nutrition and other diseases are also likely to have an impact. Aim(s): To determine the safety, reactogenicity and immunogenicity of the inactivated whole virus vaccine (Covaxin) amongst hospital-based population groups. Material(s) and Method(s): The prospective analytical study was conducted in the Department of Microbiology, Sawai Man Singh Medical College, Jaipur, Rajasthan, India, from January 2021 to March 2021.The study primarily included Healthcare Workers (HCWs) employed at SMS Medical college and attached hospitals. In-vitro quantitative IgG antibodies against SARS-CoV-2 spike Receptor Binding Domain (RBD) were measured using Chemiluminescence Immunoassay (CLIA) based Advia centaur SARS-CoV-2 IgG, manufactured by Siemens Pvt Ltd, Munich, Germany, as per manufacture's instructions. Result(s): Out of total 223 individuals, 61.88 % (138/223) showed neutralising antibody titre of >1 index value by CLIA, rest 38.12% (85/223) were non reactive i.e., titre <1 index value, after four weeks of receiving first dose of Covaxin. After 2 to 4 weeks of receiving second dose 84.30% (188/223) showed neutralising antibody titre of >1 index value by CLIA, rest 15.70% (35/223) were non reactive i.e., titre <1 index value. After receiving first dose, 100% (223/223) of the participants developed localised pain and bodyache 33.63% (75/223). None of the participants showed any anaphylactic reaction or any emergency condition just after vaccination. Conclusion(s): Covaxin is a well-tolerated vaccine, and induces good humoral response against SARS-CoV-2 with a significant rise in the neutralising antibody titres. Copyright © 2022 Journal of Clinical and Diagnostic Research. All rights reserved.

Measurement of Black Carbon in Delhi: Evidences of Regional Transport, Meteorology and Local Sources for Pollution Episodes

Measurement of particulate matter (PM) constituent such as black carbon (BC) over urban sites is critically important owing to its adverse health and climate impacts. However, the impacts associated with BC are poorly understood primarily because of the scarcity and uncertainties of measurements of BC. Here, we present BC measurement at an urban site of Delhi using a characterized continuous soot monitoring system (COSMOS) for a year-long period, i.e., from September, 2019 to August, 2020. This measurement period covers events, i.e., period of crop residue burnings from nearby states, festive events, e.g., Diwali and New Year, and first COVID-19 lockdown period. Effects of these events combining with local emissions and meteorological conditions on BC mass concentration (MBC) are investigated to find the possible cause of severe pollution levels in Delhi. Mean MBC for the complete observation period was found to be 5.02 ?? 4.40 ??g m???3. MBC showed significant seasonal as well diurnal variations. Winter season (December to February) is observed to be the most polluted season owing to increased local emissions and non-favorable meteorological conditions. Regional emission from crop burning in nearby states during October and November is the main contributing factor for increased pollution in this postmonsoon season. Furthermore, analysis reveals that cracker burning during festivals can also be considered as contributing factor to high MBC for a short period in post-monsoon and winter seasons. Significant decrease in MBC due to COVID-19 lockdown is also observed. MBC in summer and monsoon are lower as compared to other seasons but are still higher than mean MBC levels in several other urban cities of different countries. Also, the BC data obtained from nearby sites surface black carbon (SBC) are compared against the MBC to evaluate coherency among the different datasets, and discussed in detail.

Novel EHR-Derived SARS-CoV-2 Pneumonia Subphenotypes Show Divergent Clinical Outcomes with Differential Responses to Corticosteroids

Introduction: In COVID-19-related acute respiratory distress syndrome (ARDS), two distinct subphenotypes have been identified with differential outcomes and responses to corticosteroid therapy. We aimed to evaluate (1) whether clinical data can identify subgroups in a broader group of patients with SARS-CoV-2 pneumonia and (2) the extent to which corticosteroids demonstrate heterogeneity of treatment effect across such subgroups. Methods: We retrospectively studied all SARS-CoV-2 patients hospitalized for >24 hours and requiring oxygen support across 11 BJC HealthCare hospitals from June-December 2020. We excluded the initial surge (March-May 2020), as clinical care was heterogeneous and corticosteroid use low during this period. Using prespecified routinely-collected vital sign and laboratory indicator variables, we sought distinct clinical subphenotypes of SARS-CoV-2 pneumonia through latent class analysis (LCA). Across LCA subphenotypes, we evaluated the relationship between corticosteroid treatment and patient outcomes. We used multivariable logistic regression (dependent variable = composite of death/hospice) to explore treatment interaction between corticosteroid exposure and LCA subphenotype, adjusting for age and maximal SOFA score within 24 hours of admission as surrogates for indication. Results: The 3-class LCA model best fit the 1845-patient cohort (p=0.007). Class-1 (n=1456) had mean standardized values of all indicator variables near zero;Class-2 (n=235) manifested profound isolated hypoxemia;and Class-3 (n=154) displayed multiorgan failure, shock, and neutrophilia (Figure-1A). Despite representing <25% of the cohort, Classes 2 (n=109, 46%) and 3 (n=70, 46%) comprised >50% of the primary outcome (vs Class-1: n=151, 10%;p<0.001). Corticosteroids were more frequently administered in Class-2 (n=215, 91%) than in Class-1 (n=1071, 74%) or Class-3 (n=110, 71%, p<0.001;Figure-1B). Adjusted analyses demonstrated interaction between LCA class and corticosteroid treatment for the primary outcome (Class-1, p=0.003;Class-3, p=0.002). Corticosteroids were associated with increased adjusted odds for the primary outcome in Class-1 (aOR 2.11, 95% CI 1.33-3.50, p=0.002) and decreased adjusted odds for the primary outcome in Class-3 (aOR 0.44, 95% CI 0.19-0.98, p=0.048). Class-2 showed no outcome differences between the corticosteroid and noncorticosteroid groups (aOR 1.03, 95% CI 0.38-2.81, p=0.95). Conclusions: Three distinct subphenotypes of SARS-CoV-2 pneumonia demonstrate different clinical outcomes and corticosteroid response. No clear effect was seen among patients with isolated hypoxemia, whereas those with multiorgan failure appeared to benefit. Our findings suggest that among hospitalised patients in our healthcare system, corticosteroid therapy was associated with increased risk of harm. Prospective studies are needed to evaluate the efficacy of corticosteroids in SARS-CoV-2 pneumonia in predictively-enriched trials.

Hyperinflammatory ARDS Is Characterized by Interferon-High, Neutrophil-Predominant Inflammation in the Lower Respiratory Tract

Background: Latent class analyses in patients with acute respiratory distress syndrome (ARDS) have identified “hyper-inflammatory” and “hypo-inflammatory” phenotypes with divergent clinical outcomes and treatment responses. ARDS phenotypes are defined using plasma biomarkers and clinical variables. It is currently unknown if these phenotypes have distinct pulmonary biology and if pre-clinical models of disease replicate the biology of either phenotype. Methods: 45 subjects with ARDS (Berlin Definition) and 5 mechanically ventilated controls were selected from cohorts of mechanically ventilated patients at UCSF and ZSFG. Patients with COVID-19 were excluded from this analysis. A 3-variable classifier model (plasma IL-8, protein C, and bicarbonate;Sinha 2020) was used to assign ARDS phenotypes. Tracheal aspirate (TA) RNA was analyzed using established bulk and single-cell sequencing pipelines (Langelier 2018, Sarma 2021). Differentially expressed (DE) genes were analyzed using Ingenuity Pathway Analysis (IPA). Microbial community composition was analyzed with vegan. Fgsea was used to test for enrichment of gene sets from experimental ARDS models in genes that were differentially expressed between each phenotype and mechanically ventilated controls. Results: Bulk RNA sequencing (RNAseq) was available from 29 subjects with hypoinflammatory ARDS and 10 subjects with hyperinflammatory ARDS. 2,777 genes were differentially expressed between ARDS phenotypes. IPA identified several candidate upstream regulators of gene expression in hyperinflammatory ARDS including IL6, TNF, IL17C, and interferons (Figure 1A). 2,953 genes were differentially expressed between hyperinflammatory ARDS and 5 ventilated controls;in contrast, only 243 genes were differentially expressed between hypoinflammatory ARDS and controls, suggesting gene expression in the hypoinflammatory phenotype was more heterogeneous. Gene sets from experimental models of acute lung injury were enriched in hyperinflammatory ARDS but not in hypoinflammatory ARDS (Figure 1B). Single cell RNA sequencing (scRNAseq) was available from 6 additional subjects with ARDS, of whom 3 had hyperinflammatory ARDS. 14,843 cells passed quality control filters. Hyperinflammatory ARDS subjects had a markedly higher burden of neutrophils (Figure 1C), including a cluster of stressed neutrophils expressing heat shock protein RNA that was not present in hypoinflammatory ARDS. Expression of a Th1 signature was higher in T cells from hyperinflammatory ARDS. Differential expression analysis in macrophages identified increased expression of genes associated with mortality in a previous study of ARDS patients (Morell 2019). Conclusions: The respiratory tract biology of ARDS phenotypes is distinct. Hyperinflammatory ARDS is characterized by neutrophilic inflammation with distinct immune cell polarization. Transcriptomic profiling identifies candidate preclinical disease models that replicate gene expression observed in hyperinflammatory ARDS.

Revisiting the ROSE Trial - Why Did Participants with ARDS Identified as Hispanic/Latino Have Lower Mortality When Randomized to Receive Early Neuromuscular Blockade?

Rationale: The ROSE trial was a multicenter unblinded randomized clinical trial comparing early neuromuscular blockade (NMB) to usual care in patients with moderate to severe ARDS (NEJM 2019). This trial (n=1006) was stopped early for futility yet a subgroup analysis found that among Hispanic/Latino participants the NMB intervention group had a significantly lower mortality (32%) compared to those in the control group (53.7% p=0.02 for interaction). To evaluate potential contributors to these differences we compared baseline clinical and biological characteristics among Hispanic/Latino participants in the intervention vs control group. Methods: We compared demographics primary ARDS risk factor illness severity ventilatory parameters comorbidities and plasma biomarkers at baseline between the NMB intervention and control group for all 118 Hispanic/Latino patients recruited to the ROSE trial (11.6% of the trial population). We used multiple logistic regression to examine whether the mortality difference by treatment group would persist after controlling for the factors that differed significantly between groups. Results: At baseline Hispanic/Latino participants randomized to the control group had greater disease severity scores (APACHE III SOFA;p<0.05 for both) and a higher prevalence of shock (p=0.01) compared to those randomized to the intervention. There were no significant differences between groups in causes of lung injury or baseline ventilatory parameters. In an unadjusted logistic regression model the NMB intervention was significantly associated with mortality (OR 0.42;95%CI 0.20-0.89 p=0.02). The NMB intervention was no longer significantly associated with mortality when adjusting for severity of by illness by either SOFA score (OR 0.53;95%CI 0.24-1.20 p=0.13), APACHE III (OR 0.51, 95%CI 0.20- 1.30 p=0.16) or shock as defined by the need for vasopressors (OR 0.48, 95%CI 0.22-1.03, p=0.06). Hispanic/Latino participants in the control group had significantly higher interleukin-8 (p=0.02) and lower bicarbonate (p=0.045) than those in the intervention group. Conclusion: Together these clinical and biomarker data support the conclusion that the lower mortality associated with NMB in the Hispanic/Latino subgroup may have been partially due to baseline imbalances in systemic severity of illness. This finding underscores the need to cautiously interpret apparent treatment benefits within small subgroups. The COVID-19 pandemic has highlighted ethnic and racial disparities in ARDS. Future trials will benefit from increased representation of populations that are disproportionately affected to minimize the impact of spurious findings related to small sample sizes while creating more statistical power to prospectively address disparities.

Identification and Characterization of Novel Mutants of Nsp13 Protein among Indian SARS-CoV-2 Isolates

Background: SARS-CoV-2, the causative agent of COVID-19, has mutated rapidly, enabling it to adapt and evade the immune system of the host. Emerging SARS-CoV-2 variants with crucial mutations pose a global challenge in the context of therapeutic drugs and vaccines developing globally. There are currently no specific therapeutics or vaccines available to combat SARS-CoV-2 devastation. Concerning this, the current study aimed to identify and characterize the mutations found in the Nsp13 of SARS-CoV-2 in Indian isolates. Methods: In the present study, the Clustal omega tool was used for mutational analysis. The impact of mutations on protein stability, flexibility, and function was predicted using the DynaMut and PROVEAN tools. Furthermore, B-cell epitopes contributed by Nsp13 were identified using various predictive immunoinformatic tools. Results: Non-structural protein Nsp13 sequences from Indian isolates were analyzed by comparing them with the firstly reported Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) protein sequence in Wuhan, China. Out of 825 Nsp13 protein sequences, a total of 38 mutations were observed among Indian isolates. Our data showed that mutations in Nsp13 at various positions (H164Y, A237T, T214I, C309Y, S236I, P419S, V305E, G54S, H290Y, P53S, A308Y, and A308Y) have a significant impact on the protein's stability and flexibility. Moreover, the impact of Nsp13 mutations on protein function was predicted based on the PROVEAN score that indicated 15 mutants as neutral and 23 mutants as deleterious effects. Immunological parameters of Nsp13, such as antigenicity, allergenicity, and toxicity, were evaluated to predict the potential B-cell epitopes. The predicted peptide sequences were correlated with the observed mutants. Our predicted data showed that there are seven high-rank linear epitopes as well as 18 discontinuous B-cell epitopes based on immunoinformatic tools. Moreover, it was observed that out of the total 38 identified mutations among Indian SARS-CoV-2 Nsp13 protein, four mutant residues at positions 142 (E142), 245 (H245), 247 (V247), and 419 (P419) were localised in the predicted B cell epitopic region. Conclusion: Altogether, the results of the present in silico study might help to understand the impact of the identified mutations in Nsp13 protein on its stability, flexibility, and function. © 2022 Kumari et al.

Comparison of Different RNA Extraction Methods for Detection of SARS-Cov-2by RT-PCR

Background: Corona virus disease 2019 is a highly infectious disease which is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2. SARS-CoV-2 is transmitted from person to person mainly by respiratory droplets and aerosols as well as by direct or indirect contact. Aims and objective: To compare different RNA extraction methods for detection of SARSCov-2 RNA from nasopharyngeal and oropharyngeal swabs using three different methods which are based on different techniques. Material and methods: This analytical observational study was conducted in the department of Microbiology, Sawai Man Singh Medical College Jaipur, Rajasthan from December 2020 to January 2021. We selected 200 confirmed positive (extracted by Easy Mag automated system) (remnant) samples showing a wide range of different Ct values and 20 confirmed negative samples stored in Viral Transport Media VTM for this study. In order to compare quality of three extractions methods, all samples were aliquoted separately for each extraction technique. (1) Extraction by manual method (spin column base): was done by as per manufacturer’s instructions. (2) Extraction by QIA cube HT (vaccum column base): was done by as per manufacturer’s instructions. (3) Extraction by Perkins Elmer chemagic 360: (magnetic beads based). Result: A panel consisting of 200 Covid-19 positive and 20 Covid-19 negative samples were extracted by three methods (i.e. Manual column based, automated column-based and automated magnetic beads-based method). The extracted material/elutes were put for realtime RT-PCR assay for the detection of SARS CoV-2 RNA. There was no major difference seen in individual samples’ ct values between three extraction system. CONCLUSION: In conclusion, we recommended all three RNA extraction methods (i.e. magnetic beads & silica column-based) are interchangeable in a diagnostic workflow for the SARS CoV-2 by RTPCR and can be taken into account for SARS CoV-2 detection in possible future shortage of one kit or times of crisis in such pandemic time.

Seasonal Variation of Influenza A Virus and Its Subtypes in Patients Attending Tertiary Care Hospital

Background:Influenza is an important respiratory infection, causing 250,000 to 500,000 deaths annually. Influenza virus A is the most virulent and associated with winter epidemics in temperate regions, more persistent transmission in the tropics, and occasional large-scale global pandemics. But, there is variability in the pattern, and the H1N1 pandemic of 2009-2010 was unusually with a large spike in spring and a sharp decline continuing throughout winter. Varying in pattern is due to antigenic shift and drift and reassortment of the virus. Methods:A prospective study was carried out in Advance Basic Sciences & Clinical Research Lab, Department of Micro- biology in SMS Medical College & Hospital, Jaipur for diagnosis of Influenza A virus as well as subtyping was done using RT-PCR technique over 1 year period (July 2019 to June 2020) and demographic data was noted. Results:Total of 7213 samples were tested, out of which 498 (6.90%) were positive for Influenza A which is less from the previous year’s 22.46%. Out of total positive cases Influenza a (H1N1) pdm09 was 24.9% and InfA H3N2 was 75.10%. InfA H3N2 was the prominent circulating strain in all months while Influenza a (H1N1) pdm09 was prominent strain pre- vious year. Majority of positive cases were found in March 2020 (43.17%), September 2019 (28.51%). Most of these cases 36.14% were from age group between 20 to 40 years. Conclusions: A decline in the positivity of influenza infection compared to last year is seen which could be in part due to circulation of SARS COV 2 and measures of prevention undertaken by community to prevent it. Demographic parame- ters and seasonal variation of Influenza A virus give ideas to create awareness and to improve control strategies to mini- mize the morbidity, mortality and spread of disease.

Extraction Free Rapid Detection of SARS-CoV-2 from Oropharyngeal/Nasopharyngeal Swabs by Real Time PCR

Introduction: The emergence of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) pandemic has been troublesome particularly for developing countries that lack infrastructure and capacities to produce the kits locally. Simplification of the method can increase diagnostic efficiency which can benefit patients and help in infection control, consequently saving time and lives. Aim: To evaluate the diagnostic value of four methods (that omit extraction step) for detection of SARS-CoV-2 against the traditional extraction method. Materials and Methods: This was a cross-sectional analysis for evaluating diagnostic accuracy of four methods for detection of SARS-CoV-2 by real-time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCR), conducted in the Department of Microbiology, SMS Medical College, Jaipur, Rajasthan, India, in October 2020. Ninety four SARS-CoV-2 RT-PCR positive samples and 20 negative samples were taken for this study. Automated extraction system was used for Ribonucleic Acid (RNA) extraction and four different approaches were compared to the traditional extraction method for detection of SARS-CoV-2 by RT-PCR. Data was entered and analysed using Statistical Package for the Social Sciences (SPSS) statistical software version 24.0. Results: The automated RNA extraction method was compared to the method of direct addition of samples with (Heat processed Direct Viral transport medium Sample (HDVS)) and without heating (Direct Viral transport medium Sample (DVS)), directs addition of diluted (1:5) sample with (Heat processed diluted VTM sample (HdVS)) and without heating (Diluted VTM sample (dVS)) as well as after addition of Proteinse K (PK) to the diluted samples that came either negative/invalid. Out of four methods, the HdVS method gave the best results, considering extraction with Perkin Elmer as standard, this method showed sensitivity of 96.74%, specificity of 100%. Conclusion: In current pandemic, molecular testing is critically challenged by the limited supplies of reagents of nucleic acid extraction alternative method like diluting and heating of Viral Transport Media (VTM) samples and using them directly as elutes serve as an easy, fast and inexpensive alternative.