Divalent siRNAs are bioavailable in the lung and efficiently block SARS-CoV-2 infection.

The continuous evolution of SARS-CoV-2 variants complicates efforts to combat the ongoing pandemic, underscoring the need for a dynamic platform for the rapid development of pan-viral variant therapeutics. Oligonucleotide therapeutics are enhancing the treatment of numerous diseases with unprecedented potency, duration of effect, and safety. Through the systematic screening of hundreds of oligonucleotide sequences, we identified fully chemically stabilized siRNAs and ASOs that target regions of the SARS-CoV-2 genome conserved in all variants of concern, including delta and omicron. We successively evaluated candidates in cellular reporter assays, followed by viral inhibition in cell culture, with eventual testing of leads for in vivo antiviral activity in the lung. Previous attempts to deliver therapeutic oligonucleotides to the lung have met with only modest success. Here, we report the development of a platform for identifying and generating potent, chemically modified multimeric siRNAs bioavailable in the lung after local intranasal and intratracheal delivery. The optimized divalent siRNAs showed robust antiviral activity in human cells and mouse models of SARS-CoV-2 infection and represent a new paradigm for antiviral therapeutic development for current and future pandemics.

DNA aptamers inhibit SARS-CoV-2 spike-protein binding to hACE2 by an RBD- independent or dependent approach.

Objective: Nobody knows when the COVID-19 pandemic will end or when and where the next coronavirus will outbreak. Therefore, it is still necessary to develop SARS-CoV-2 inhibitors for different variants or even the new coronavirus. Since SARS-CoV-2 uses its surface spike-protein to recognize hACE2, mediating its entry into cells, ligands that can specifically recognize the spike-protein have the potential to prevent infection. Methods: We have recently discovered DNA aptamers against the S2-domain of the WT spike-protein by exploiting the selection process called SELEX. After optimization, among all candidates, the aptamer S2A2C1 has the shortest sequence and the best binding affinity toward the S2-protein. More importantly, the S2A2C1 aptamer does not bind to the RBD of the spike-protein, but it efficiently blocks the spike-protein/hACE2 interaction, suggesting an RBD-independent inhibition approach. To further improve its performance, we conjugated the S2A2C1 aptamer with a reported anti-RBD aptamer, S1B6C3, using various linkers and constructed hetero-bivalent fusion aptamers. Binding affinities of mono and fusion aptamers against the spike-proteins were measured. The inhibition efficacies of mono and fusion aptamers to prevent the hACE2/spike-protein interaction were determined using ELISA. Results: Anti-spike-protein aptamers, including S2A2C1 and S1B6C3-A5-S2A2C1, maintained high binding affinity toward the WT, Delta, and Omicron spike-proteins and high inhibition efficacies to prevent them from binding to hACE2, rendering them well-suited as diagnostic and therapeutic molecular tools to target SARS-CoV-2 and its variants. Conclusions: Overall, we discovered the anti-S2 aptamer, S2A2C1, which inhibits the hACE2/spike-protein interaction via an RBD-independent approach. The anti-S2 and anti-RBD aptamers were conjugated to obtain the fusion aptamer, S1B6C3-A5-S2A2C1, which recognizes the spike-protein by an RBD-dependent approach. Our strategies, which discovered aptamer inhibitors targeting the highly conserved S2-protein, as well as the design of fusion aptamers, can be used to target new coronaviruses as they emerge.

Awareness and understanding of COVID-19 among pregnant woman in Northern India.

BACKGROUND: The severe acute respiratory syndrome coronavirus (SARS-CoV-2) has become a public health concern worldwide. It is important for pregnant women to know about the mode of transmission, symptoms, and preventive measures against COVID-19. The aim of this study was to evaluate the awareness and outlook of pregnant women and practical measures taken by them against COVID-19. MATERIALS AND METHODS: This cross-sectional questionnaire-based study was conducted for 1 month (June 10, 2020- July 10, 2020) in the Department of Obstetrics and Gynecology, All India Institute of Medical Sciences, Jodhpur, Rajasthan, India. A prevalidated questionnaire was given to 109 pregnant women to assess the knowledge, attitude, and practice acquired against COVID-19 infection. The Chi-square test or Fischer's exact test was used to compare categorical data. RESULTS: Among 109 participants, 103 (94.5%) had good knowledge, 4 (3.7%) had average knowledge and 2 (1.8%) had poor knowledge about COVID-19. Majority of them had a positive attitude for the protective measures taken for the prevention of disease. CONCLUSIONS: Since there is no valid treatment for COVID-19, prevention is the only key to curve this infection. In the present study, 94.5% pregnant women had overall good knowledge score about the mode of transmission, symptoms, and preventive measures against COVID-19.

Wilms tumor with Mulibrey Nanism: A case report and review of literature.

BACKGROUND: Mulibrey-Nanism (Muscle-liver-brain-eye Nanism = dwarfism; MUL) is a rare genetic syndrome. The underlying TRIM37 mutation predisposes these children to develop tumors frequently. In the largest published series of MUL, 8% patients were reported to develop Wilms tumor (WT). The published literature lacks data regarding the best treatment protocol and outcome of this cohort of children with WT and MUL. We report here a 2-year-old boy with WT and MUL and present a review of literature on WT in MUL. CASE: Our patient had associated cardiac problems of atrial septal defect, atrial flutter and an episode of sudden cardiac arrest. We managed him successfully with chemotherapy, surgery and multi-speciality care. He is alive and in remission at follow-up of 6 months. CONCLUSION: A total of 14 cases (including present case) of WT have been reported in MUL and treatment details were available for six cases. They were managed primarily with surgery, chemotherapy with/without radiotherapy, and all achieved remission. The outcome data is available only for two cases, one has been followed up till 15 years post treatment for WT and other is our patient.

A universal DNA aptamer as an efficient inhibitor against spike-protein/hACE2 interactions.

A universal aptamer against spike-proteins of diverse SARS-CoV-2 variants was discovered via DNA SELEX towards the wild-type (WT) spike-protein. This aptamer, A1C1, binds to the WT spike-protein or other variants of concern such as Delta and Omicron with low nanomolar affinities. A1C1 inhibited the interaction between hACE2 and various spike-proteins by 85-89%. This universal A1C1 aptamer can be used to design diagnostic and therapeutic molecular tools to target SARS-CoV-2 and its variants.

COVID-19 in children with severe aplastic anemia

Introduction Coronavirus disease 2019 (COVID-19) affects children but mostly has mild course. There is meagre published data on the impact of COVID-19 illness in children with Severe Aplastic anemia (SAA). We describe our experience of managing COVID-19 in children with SAA. Method Three children of SAA who developed SARS-CoV-2 infection are included in this study. Results Patient 1 was post Immunosuppressive therapy (IST) for SAA and had an asymptomatic course and uneventful recovery. Patient 2 was several months post IST with no response and had an asymptomatic COVID-19 illness but had delayed viral clearance, however he succumbed to bacterial sepsis soon after. Patient 3 was awaiting IST and while he contracted severe acute respiratory syndrome-Coronavirus-2 (SARS-CoV-2) infection, he had symptomatic COVID-19 illness followed by bacterial and fungal sepsis to which he succumbed. Conclusion: COVID-19 in children with SAA can be mild to fatal course and virus may have delayed clearance. It can lead to delay in therapy of SAA.

Outcomes of COVID-19 in children with cancer – Report from the Indian Pediatric Oncology Group (InPOG) COVID-19 registry in India

Introduction The clinical outcomes of COVID-19 infection in children with cancer have been variable worldwide. Therefore, we aimed to collect data from all regions in India through a national collaborative study and identify factors that cause mortality directly related to COVID-19 infection. Methods Data was collected prospectively on children across India on cancer therapy and diagnosed with COVID-19 infections from 47 centers from April 2020 to October 2021. Information was recorded on the demographics, the number of children that required intervention, and the outcome of the infection. In addition, we analyzed the impact of the delta variant in 2021. Results A total of 659 children were studied, of whom 64% were male and 36% were female. The data from the eastern region was sparse, and this was a collection bias. COVID-19 infection was predominantly seen in children less than five years. The delta variant had a higher impact in the southern region, and this was statistically significant. Of the 659 children, 30 children died (4.5%), however only 7 of the deaths were directly attributed to COVID-19 infection (1%). Conclusion The study reports the largest nationally representative cohort of children with cancer and COVID-19 to date in India. We identified demographic and clinical factors associated with increased all-cause mortality in patients with cancer. Complete characterization of the cohort has provided further insights into the effects of COVID-19 on cancer outcomes. The low mortality allows us to recommend that specific cancer treatments be continued without delays in therapy.

Rhino-orbital mucormycosis: Our experiences with clinical features and management in a tertiary care center

Objective: To determine the prevalence, risk factors, and elaborate our experiences with diagnosis and treatment of patients with mucormycosis, enabling a better understanding of the disease and its management. Methods: This is a case series of patients with Covid-19 associated with Rhino-orbital-cerebral mucormycosis, managed in our tertiary care center from April 2021 to June 2021. Results: Six cases of Covid-19 associated with Rhino-orbital-cerebral mucormycosis have been analyzed in the study. The mean age of patients was 40.67 years with a male preponderance (83.3%). The most common complaint was headache (100%), while a minority (33%) came with ocular complaints. All the patients either had a previous history of diabetes mellitus or developed increased blood sugar levels following Covid infection, and were kept on insulin to control their blood sugar levels. 4 patients (66.67%) had a history of corticosteroid use during Covid-19 hospitalization. Treatment included intravenous liposomal Amphotericin B (100%), functional endoscopic sinus surgery (66.67%), maxillectomy (33.33%) and transcutaneous retrobulbar liposomal Amphotericin B (33.33%). Amphotericin B induced nephrotoxicity, which was seen in 1 patient (16.67%). Mortality occurred in only one patient (16.67%), 25 days following successful surgery. Conclusion: Diabetes Mellitus is the most important predisposing factor for the development of Covid-19 associated Rhino-orbital-cerebral mucormycosis. Early presentation, prompt diagnosis and timely initiation of treatment with liposomal Amphotericin B and surgical debridement along with strict blood sugar control can lead to a favorable outcome. However, regular follow-up and monitoring of serum electrolytes and kidney profile must be ensured for such patients.

Challenges and actions to the environmental management of bio-medical waste during COVID-19 pandemic in India

The outbreak of COVID-19 has engendered a global health crisis along with diverse impacts on economy, society and environment. Efforts to combat this pandemic have also significantly shot-up the quantity of Bio-medical Waste (BMW) generation. Safe disposal of large quantity of BMW has been gradually posing a major challenge. BMW management is mostly implemented at municipal level following regulatory guidelines defined by respective states and the Union. This article is a narrative of the status of BMW generation, management and regulation in India in the context of COVID-19 crisis. The article is based on comparative analysis of data on BMW generation and management from authentic sources, a systematic literature review and review of news reports. In the current pandemic situation where media has been playing a significant role in highlighting all the concerns related to COVID-19 spread and management. Assessing the ground situation regarding effectiveness of prevailing BMW management facilities, requirement and suggestions can provide insights to the subject with policy implications for India and countries as well. The discussion has been built on different dimensions of BMW management during the pandemic including existing infrastructures, capacity utilisation, policy guidelines, operational practices and waste-handlers aspects. The results on state-wise analysis of reported BMW quantity and active COVID-19 patients also reveal some non-linear relationship between the two variables. Delhi, the National Capital is situated at a better position in terms of BMW management as compared to other studied states. The findings are expected to provide valuable insights to the policy makers and other relevant authorities to evaluate adequateness as well as efficiency quotients of entire BMW management landscape. Some of the critical observations of this article are also expected to offer impetus for enhancing national disaster preparedness in future.

"Molecular Masks" for ACE2 to Effectively and Safely Block SARS-CoV-2 Virus Entry.

Coronavirus Disease 2019 (COVID-19) remains a global health crisis, despite the development and success of vaccines in certain countries. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19, uses its spike protein to bind to the human cell surface receptor angiotensin-converting enzyme 2 (ACE2), which allows the virus to enter the human body. Using our unique cell screening technology, we identified two ACE2-binding peptoid compounds and developed dimeric derivatives (ACE2P1D1 and ACE2P2D1) that effectively blocked spike protein-ACE2 interaction, resulting in the inhibition of SARS-CoV-2 pseudovirus entry into human cells. ACE2P1D1 and ACE2P2D1 also blocked infection by a D614G mutant pseudovirus. More importantly, these compounds do not decrease ACE2 expression nor its enzyme activity (which is important in normal blood pressure regulation), suggesting safe applicability in humans.

SARS-CoV-2: Understanding the Transcriptional Regulation of ACE2 and TMPRSS2 and the Role of Single Nucleotide Polymorphism (SNP) at Codon 72 of p53 in the Innate Immune Response against Virus Infection.

Human ACE2 and the serine protease TMPRSS2 of novel SARS-CoV-2 are primary entry receptors in host cells. Expression of these genes at the transcriptional level has not been much discussed in detail. The ISRE elements of the ACE2 promoter are a binding site for the ISGF3 complex of the JAK/STAT signaling pathway. TMPRSS2, including IFNß, STAT1, and STAT2, has the PARP1 binding site near to TSS either up or downstream promoter region. It is well documented that PARP1 regulates gene expression at the transcription level. Therefore, to curb virus infection, both promoting type I IFN signaling to boost innate immunity and prevention of virus entry by inhibiting PARP1, ACE2 or TMPRSS2 are safe options. Most importantly, our aim is to attract the attention of the global scientific community towards the codon 72 Single Nucleotide Polymorphism (SNP) of p53 and its underneath role in the innate immune response against SARS-CoV-2. Here, we discuss codon 72 SNP of human p53's role in the different innate immune response to restrict virus-mediated mortality rate only in specific parts of the world. In addition, we discuss potential targets and emerging therapies using bioengineered bacteriophage, anti-sense, or CRISPR strategies.

The Impact of IoT in Healthcare: Global Technological Change & The Roadmap to a Networked Architecture in India.

The digitization of data including health data (referred to as Internet of Things-IoT in Healthcare) and its usage in delivery of healthcare has been growing rapidly across the world. The COVID-19 pandemic has been a pivot for exponential growth of IoT in healthcare. Several rapidly evolving technologies are converging to influence the trajectory of IoT in healthcare. There are several challenges in technology development, healthcare delivery as well as issues related to privacy of data, digital divide, role of government and other stakeholders, behaviour and adoption by medical doctors and hospitals. The review article provides an overview of the emergence of IoT in healthcare globally, the intricacies of different factors impinging its current status and recommends policy intervention for an optimal roadmap of IoT in healthcare in the Indian context.

Awareness and experience of health-care workers during coronavirus disease 2019 pandemic.

BACKGROUND: During the 2nd week of July 2020, the coronavirus disease 2019 (COVID 19) infection spreading in the community. Now more than 15 lakhs peoples have been infected in India, out of the 26816 patients were deceased. COVID 19 outbreaks become an additional hazard to the health-care workers (HCWs), leading to fatigue, anxiety, depression and fear of death. The objective of this questionnaire-based study is to know about the knowledge of HCWs about COVID 19, their experiences while dealing with the disease, and the protective measures taken to prevent the infection. MATERIALS AND METHODS: A cross-sectional, questionnaire-based study was conducted for 1 month starting from the 2nd week of June 2020 after getting institutional ethical clearance. This study included 240 HCWs posted in the medical and surgical Departments of All India Institute of Medical Sciences, Jodhpur, Rajasthan. This questionnaire was prepared in online Google forms and required 2 min to complete. Mean, median, range, and standard deviation were used to describe the continuous variables, and percentages were used to describe the categorical data. RESULTS: Among 240 HCWs, 79.16% (n = 190) participants have good knowledge and adapted good precautions (score 15-23) for COVID 19 infection. Rest 20% (n = 48) and 0.8% (n = 2) participants has average (score 8-14) or poor knowledge (score <8) with adaptation of average or poor precautionary measures against COVID 19, respectively. CONCLUSION: This study concluded that we have the requirement of more educational training programs for awareness of HCWs and precautionary measures against COVID 19. Thereby, HCWs can improve their knowledge and participate in this fight against COVID 19 with more efficiency and confidence.

Estimation of real-infection and immunity against SARS-CoV-2 in Indian populations (preprint)

Infection born by Coronavirus SARS-CoV-2 has swept the world within a time of a few months. It has created a devastating effect on humanity with social and economic depressions. Europe and America were the hardest hit continents. India has also lost several lives, making the country fourth most deadly worldwide. However, the infection and death rate per million and the case fatality ratio in India were substantially lower than many of the developed nations. Several factors have been proposed including the genetics. One of the important facts is that a large chunk of Indian population is asymptomatic to the SARS-CoV-2 infection. Thus, the real infection in India is much higher than the reported number of cases. Therefore, the majority of people are already immune in the country. To understand the dynamics of real infection as well as level of immunity against SARS-CoV-2, we have performed antibody testing (serosurveillance) in the urban region of fourteen Indian districts encompassing six states. In our survey, the seroprevalence frequency varied between 0.01-0.48, suggesting high variability of viral transmission among states. We also found out that the cases reported by the Government were several fold lower than the real infection. This discrepancy is majorly driven by a higher number of asymptomatic cases. Overall, we suggest that with the high level of immunity developed against SARS-CoV-2 in the majority of the districts, it is less likely to have a second wave in India.