Signature Inflammatory Cytokine panel: IL-10, IL-6, VEGF and IL-8 in Covid-19 (preprint)

Objective We aimed to assess a battery of inflammatory cytokines in SARS-CoV-2 patients to determine the cytokines of prognostic and/ predictive relevance in Covid-19.Methods In a cohort of total 100 SARS-CoV-2 patients (RT-PCR confirmed) hospitalized in associated SMHS hospital of GMC Srinagar, Kashmir (North India), the level of a battery of cytokines IL-6, IL-8, IL-10, IL-1α and VEGF, TNF-α and ferritin, were estimated by Enzyme Linked Immunosorbent Assay ( ELISA) on Multimode Microplate reader.Result The deranged levels of these cytokines were mostly found in patients > 60 years of age with cough and pneumonia as the most common symptoms. Correlation analysis revealed significant association between interleukin's IL-6, IL-8 and disease severity (p = 0.002) (p = 0.007) and poor disease outcome (p = 0.04), (p = 0.009) respectively. Similar association was also found between decreased levels of VEGF and poor disease outcome (p = 0.02). Further ROC analysis, univariant and multivariant (after adjusting for age, gender and other inflammatory markers), revealed increased IL-10 (AUC = 0.72) and IL-6 (AUC = 0.70) as independent markers of both disease severity(p = 0.02) (p = 0.01) and disease outcome (P = 0.03) (p = 0.02) and decreased VEGF (AUC = 0.69) as independent marker of disease outcome only (p = 0.03). Significant association of cough with IL-8 levels (p = 0.01) and of diabetes with raised ferritin levels (p = 0.01) with very high ferritin levels (> 1500ng/ml) as indicator of those that are likely to develop hyperinflammatory phenotype was found in SARS-CoV-2 patients.Conclusion We conclude ‘IL-6, IL10, VEGF and IL-8’ as the signature inflammatory cytokine panel in Covid-19. An increased IL-10, IL-6 levels proved to be equally significant independent prognosticators of Covid − 19 severity and predictors of poor disease outcome and decreased VEGF level as predictors of poor disease outcome in SARS-CoV-2 patients. Testing of the signature inflammatory cytokine panel is, therefore, recommended for optimal clinical decision making in Covid-19.

Impact of Age and Clinico-Biochemical Parameters on Clinical severity of SARS-CoV-2 Infection.

INTRODUCTION: The surge in novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection leading to COrona VIrus Disease-2019 (COVID-19) has overwhelmed the health system. To help health-care workers and policy makers prioritize treatment and to decrease the burden on health systems caused by COVID-19, clinical severity along with various clinico-biochemical parameters was evaluated by designing a cross-sectional study comprising of 236 SARS-CoV-2 infected individuals from Kashmir valley, India. METHODS: Briefly, Real time Polymerase chain reaction (RT-PCR) was used for the confirmation of SARS-CoV-2 infection. The principles of spectrophotometry and chemiluminescent micro particle immunoassay (CMIA) were employed to estimate the levels of Glucose, TSH and 25-hydroxy vitamin D levels in serum of infected patients. RESULTS: A total of 236 patients infected with SARS-CoV-2 were taken for this cross-sectional study. Patients with COVID-19 had a male predominance (72.9% vs 27.1%) and higher prevalence of 25-hydroxy vitamin D deficiency (72.0% vs 28.0%) with a mean 25-hydroxy vitamin D levels of 24.0±13.9 in ng/ml. We observed a varied clinical spectrum of SARS-CoV-2 infection with 36.4%, 23.7% and 29.7% patients having mild, moderate and severe disease respectively. We observed that severity of SARS-CoV-2 infection was significantly associated with older age group, hypertension, low TSH levels and 25-hydroxy vitamin D deficiency. DISCUSSION/CONCLUSION: We conclude that not only old age but hypertension and low levels of TSH and 25 Hydroxy Vitamin D Levels could significantly lead to clinical severity of SARS-CoV-2 infection.

Seroprevalence of SARS-CoV-2-specific anti-spike IgM, IgG, and anti-nucleocapsid IgG antibodies during the second wave of the pandemic: A population-based cross-sectional survey across Kashmir, India.

Background: Within Kashmir, which is one of the topographically distinct areas in the Himalayan belt of India, a total of 2,236 cumulative deaths occurred by the end of the second wave. We aimed to conduct this population-based study in the age group of 7 years and above to estimate the seropositivity and its attributes in Kashmir valley. Methods: We conducted a community-based household-level cross-sectional study, with a multistage, population-stratified, probability-proportionate-to-size, cluster sampling method to select 400 participants from each of the 10 districts of Kashmir. We also selected a quota of healthcare workers, police personnel, and antenatal women from each of the districts. Households were selected from each cluster and all family members with age 7 years or more were invited to participate. Information was collected through a standardized questionnaire and entered into Epicollect 5 software. Trained healthcare personnel were assigned for collecting venous blood samples from each of the participants which were transferred and processed for immunological testing. Testing was done for the presence of SARS-CoV-2-specific anti-spike IgM, IgG antibodies, and anti-nucleocapsid IgG antibodies. Weighted seropositivity was estimated along with the adjustment done for the sensitivity and specificity of the test used. Findings: The data were collected from a total of 4,229 participants from the general population within the 10 districts of Kashmir. Our results showed that 84.84% (95% CI 84.51-85.18%) of the participants were seropositive in the weighted imputed data among the general population. In multiple logistic regression, the variables significantly affecting the seroprevalence were the age group 45-59 years (odds ratio of 0.73; 95% CI 0.67-0.78), self-reported history of comorbidity (odds ratio of 1.47; 95% CI 1.33-1.61), and positive vaccination history (odds ratio of 0.85; 95% CI 0.79-0.90) for anti-nucleocapsid IgG antibodies. The entire assessed variables showed a significant role during multiple logistic regression analysis for affecting IgM anti-spike antibodies with an odds ratio of 1.45 (95% CI 1.32-1.57) for age more than 60 years, 1.21 (95% CI 1.15-1.27) for the female gender, 0.87 (95% CI 0.82-0.92) for urban residents, 0.86 (95% CI 0.76-0.92) for self-reported comorbidity, and an odds ratio of 1.16 (95% CI 1.08-1.24) for a positive history of vaccination. The estimated infection fatality ratio was 0.033% (95% CI: 0.034-0.032%) between 22 May and 31 July 2021 against the seropositivity for IgM antibodies. Interpretation: During the second wave of the SARS-CoV-2 pandemic, 84.84% (95% CI 84.51-85.18%) of participants from this population-based cross-sectional sample were seropositive against SARS-CoV-2. Despite a comparatively lower number of cases reported and lower vaccination coverage in the region, our study found such high seropositivity across all age groups, which indicates the higher number of subclinical and less severe unnoticed caseload in the community.

Covid-19: a novel challenge to human immune genetic machinery

COVID-19 also called corona virus emerged in China in December 2019. This turned into a global pandemic in a short period of time. Covid-19 is a novel strain of corona virus that was not seen earlier in human beings. It is important to study the molecular structure of Covid-19 so as to aid in the development of therapeutic measures. Existing Covid-19 pandemic poses an extraordinary risk to health and healthcare systems worldwide. Corona viruses are made of single stranded RNA present within the coat proteins. The virus has a diameter of nearly 80–120 nm. Usually, Covid-19 presents with the signs and symptoms of respiratory illness. Cough commonly dry cough, fever, associated with myalgias and sometimes breathing difficulties due to decrease in oxygen saturation rates are also present in these patients. Some people show fever with body aches, while some are relatively asymptomatic. Corona virus is primarily transmitted in humans through respiratory route and is highly contagious. Mostly old people and those having comorbid illnesses suffer most. After invading into the human body, the virus may lead to a sequence of processes such as viral invasion, replication, and programmed cell death, that is, apoptosis. To control and prevent this viral infection, we need to study the molecular aspects of Covid-19 in detail so as to design therapeutic agents as well as for vaccine formation. The micro-RNA is defined as the single-stranded noncoding RNA molecule. They have a length of about 22 nucleotides approximately and help in the post transcriptional regulation of gene expression. Micro RNAs regulate many types of cancers in addition to Covid-19 and other infections. Viral micro RNA is a newer type of mi-RNA and controls the host cell expression and viral target genes. This was completed by inducing micro-RNA cleavage, breakdown, translation, inhibition, or other mechanisms. The micro-RNAs of Covid-19 are explained to give an authoritative means to study this novel coronavirus. These control the host cell expression and also viral target genes by inducing micro-RNA cleavage, breakdown, translation, inhibition, and also other mechanisms.

The burden of SARS-CoV-2 among healthcare workers across 16 hospitals of Kashmir, India-A seroepidemiological study.

SARS-CoV-2 pandemic has greatly affected healthcare workers because of the high risk of getting infected. The present cross-sectional study measured SARS-CoV-2 antibody in healthcare workers of Kashmir, India. METHODS: Serological testing to detect antibodies against nucleocapsid protein of SARS-CoV-2 was performed in 2003 healthcare workers who voluntarily participated in the study. RESULTS: We report relatively high seropositivity of 26.8% (95% CI 24.8-28.8) for SARS-CoV-2in healthcare workers, nine months after the first case was detected in Kashmir. Most of the healthcare workers (71.7%) attributed infection to the workplace environment. Among healthcare workers who neither reported any prior symptom nor were they ever tested for infection by nasopharyngeal swab test, 25.5% were seropositive. CONCLUSION: We advocate interval testing by nasopharyngeal swab test of all healthcare workers regardless of symptoms to limit the transmission of infection within healthcare settings.

Seroprevalence of SARS-CoV-2-specific IgG antibodies in Kashmir, India, 7 months after the first reported local COVID-19 case: results of a population-based seroprevalence survey from October to November 2020.

OBJECTIVES: We designed a population-based survey in Kashmir to estimate the seroprevalence of SARS-CoV-2-specific IgG antibodies in the general population aged 18 years and above. SETTING: The survey was conducted among 110 villages and urban wards across 10 districts in Kashmir from 17 October 2020 to 4 November 2020. PARTICIPANTS: Individuals aged 18 years and above were eligible to be included in the survey. Serum samples were tested for the presence of SARS-CoV-2-specific IgG antibodies using the Abbott SARS-CoV-2 IgG assay. PRIMARY AND SECONDARY OUTCOME MEASURES: We labelled assay results equal to or above the cut-off index value of 1.4 as positive for SARS-CoV-2-specific IgG antibodies. Seroprevalence estimates were adjusted for the sampling design and assay characteristics. RESULTS: Out of 6397 eligible individuals enumerated, 6315 (98.7%) agreed to participate. The final analysis was done on 6230 participants. Seroprevalence adjusted for the sampling design and assay characteristics was 36.7% (95% CI 34.3% to 39.2%). Seroprevalence was higher among the older population. Among seropositive individuals, 10.2% (247/2415) reported a history of COVID-19-like symptoms. Out of 474 symptomatic individuals, 233 (49.2%) reported having been tested. We estimated an infection fatality rate of 0.034%. CONCLUSIONS: During the first 7 months of the COVID-19 epidemic in Kashmir Valley, approximately 37% of individuals were infected. The reported number of COVID-19 cases was only a small fraction of the estimated number of infections. A more efficient surveillance system with strengthened reporting of COVID-19 cases and deaths is warranted.

SARS-CoV-2 Seroprevalence Among Healthcare Workers by Workplace Exposure Risk in Kashmir, India.

BACKGROUND: SARS-CoV-2 infection (COVID-19) poses a tremendous challenge to healthcare systems across the globe. Serologic testing for SARS-CoV-2 infection in healthcare workers (HCWs) may quantify the rate of clinically significant exposure in an institutional setting and identify those HCWs who are at greatest risk. METHODS: We conducted a survey and SARS-CoV-2 serologic testing among a convenience sample of HCWs from 79 non-COVID and 3 dedicated COVID hospitals in District Srinagar of Kashmir, India. In addition to testing for the presence of SARS-CoV-2-specific immunoglobulin G (IgG), we collected information on demographics, occupational group, influenza-like illness (ILI) symptoms, nasopharyngeal reverse transcription polymerase chain reaction (RT-PCR) testing status, history of close unprotected contacts, and quarantine/travel history. RESULTS: Of 7,346 eligible HCWs, 2,915 (39.7%) participated in the study. The overall prevalence of SARS-CoV-2-specific IgG antibodies was 2.5% (95% CI, 2.0%-3.1%), while HCWs who had ever worked at a dedicated COVID-19 hospital had a substantially lower seroprevalence of 0.6% (95% CI, 0.2%-1.9%). Higher seroprevalence rates were observed among HCWs who reported a recent ILI (12.2%), a positive RT-PCR (27.6%), a history of being put under quarantine (4.9%), and a history of close unprotected contact with a person with COVID-19 (4.4%). Healthcare workers who ever worked at a dedicated COVID-19 hospital had a lower multivariate-adjusted risk of seropositivity (odds ratio, 0.21; 95% CI, 0.06-0.66). CONCLUSIONS: Our investigation suggests that infection-control practices, including a compliance-maximizing buddy system, are valuable and effective in preventing infection within a high-risk clinical setting. Universal masking, mandatory testing of patients, and residential dormitories for HCWs at COVID-19-dedicated hospitals is an effective multifaceted approach to infection control. Moreover, given that many infections among HCWs are community-acquired, it is likely that the vigilant practices in these hospitals will have spillover effects, creating ingrained behaviors that will continue outside the hospital setting.

COVID-19: Diagnostics, Therapeutic Advances, and Vaccine Development.

PURPOSE OF REVIEW: Human race is currently facing the wrath of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a highly transmittable and pathogenic RNA virus, causing coronavirus disease 2019 (COVID-19), the worst ever global pandemic. Coronaviruses (CoVs) have emerged as a major public health concern. Urgent global response to COVID-19 outbreak has been to limit spread of SARS-CoV-2 via extensive monitoring and containment. Various treatment regimens have been adopted to manage COVID-19, with known drugs and drug combinations used to decrease the morbidity and mortality associated with COVID-19. Intensive research on various fronts including studying molecular and structural aspects of these viruses and unraveling the pathophysiology and mechanistic basis of COVID-19 aimed at developing effective prophylactic, therapeutic agents and vaccines has been carried out globally. RECENT FINDINGS: No approved antiviral treatment except remdesivir exists for SARS-CoV-2 till date though novel drug targets have been identified. However, worldwide frantic and competitive vaccine development pharmaceutical race has borne fruit in the form of a number of promising candidate vaccines, out of which few have already received emergency use authorization by regulatory bodies in record time. SUMMARY: This review highlights the painstaking efforts of healthcare workers and scientific community to successfully address the COVID-19 pandemic-though damage in the form of severe illness, loss of lives, and livelihood has left a serious mark. Focusing on extensive research on various therapeutic options and antiviral strategies including neutralizing antibodies, potential drugs, and drug targets, light has been shed on various diagnostic options and the amazing vaccine development process as well.

Seroprevalence of SARS-CoV-2 specific IgG antibodies in District Srinagar, northern India - A cross-sectional study.

BACKGROUND: Prevalence of IgG antibodies against SARS-CoV-2 infection provides essential information for deciding disease prevention and mitigation measures. We estimate the seroprevalence of SARS-CoV-2 specific IgG antibodies in District Srinagar. METHODS: 2906 persons >18 years of age selected from hospital visitors across District Srinagar participated in the study. We tested samples for the presence of SARS-CoV-2 specific IgG antibodies using a chemiluminescent microparticle immunoassay-based serologic test. RESULTS: Age- and gender-standardized seroprevalence was 3.6% (95% CI 2.9% to 4.3%). Age 30-69 years, a recent history of symptoms of an influenza-like-illness, and a history of being placed under quarantine were significantly related to higher odds of the presence of SARS-CoV-2 specific IgG antibodies. The estimated number of SARS-CoV-2 infections during the two weeks preceding the study, adjusted for test performance, was 32602 with an estimated (median) infection-to-known-case ratio of 46 (95% CI 36 to 57). CONCLUSIONS: The seroprevalence of SARS-CoV-2 specific IgG antibodies is low in the District. A large proportion of the population is still susceptible to the infection. A sizeable number of infections remain undetected, and a substantial proportion of people with symptoms compatible with COVID-19 are not tested.

Managing the COVID-19 Pandemic: Research Strategies Based on the Evolutionary and Molecular Characteristics of Coronaviruses.

Coronavirus disease 2019 (COVID-19), an ongoing global health emergency, is a highly transmittable and pathogenic viral infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Emerging in Wuhan, China, in December 2019, it spread widely across the world causing panic-worst ever economic depression is visibly predictable. Coronaviruses (CoVs) have emerged as a major public health concern having caused three zoonotic outbreaks; severe acute respiratory syndrome-CoV (SARS-CoV) in 2002-2003, Middle East respiratory syndrome-CoV (MERS-CoV) in 2012, and currently this devastating COVID-19. Research strategies focused on understanding the evolutionary origin, transmission, and molecular basis of SARS-CoV-2 and its pathogenesis need to be urgently formulated to manage the current and possible future coronaviral outbreaks. Current response to the COVID-19 outbreak has been largely limited to monitoring/containment. Although frantic global efforts for developing safe and effective prophylactic and therapeutic agents are on, no licensed antiviral treatment or vaccine exists till date. In this review, research strategies for coping with COVID-19 based on evolutionary and molecular aspects of coronaviruses have been proposed.