Spectrum of Fungal Infections in a Tertiary Care Centre of North India: Pre-COVID and COVID Scenario and Implications.

Background Rhino orbital mucormycosis is a rare and very aggressive entity. A sudden rise of this entity has been noticed with the insurgence of the COVID-19 pandemic both among immunocompromised and immuno-competent patients. This study was done to determine any possible correlation between these two deadly diseases. Materials and Methods This was a retrospective observational study done in the pathology department of a tertiary care center in North India over a three-year period (January 2019 - December 2021). Patient details along with relevant clinical data were retrieved from the patient's record file. Hematoxylin and eosin-stained slides of diagnosed cases were taken from the department records. Results A total of 45 patients (34 males, 11 females) were included in the study, seven of which were ophthalmic exenteration specimens. The mean age of the patients was 52.68 years. Fifteen cases showed COVID-19 reverse transcription-polymerase chain reaction (RT-PCR) positivity. Histopathology revealed the presence of mucormycosis in all the cases. There were six cases showing granuloma formation and 14 cases revealed mixed fungal infection. Optic nerve involvement was seen in six cases of exenteration specimens. Conclusions The present study showed a sudden resurgence of secondary fungal infections, especially during the second wave of the COVID-19 pandemic. Associated co-morbid conditions and injudicious use of steroids and antibiotics have been the cause of depressed immunity leading to the infections. One must be aware of such co-infections to facilitate timely medical management to reduce morbidity and mortality.

A Practical Entity Linking System for Tables in Scientific Literature (preprint)

Entity linking is an important step towards constructing knowledge graphs that facilitate advanced question answering over scientific documents, including the retrieval of relevant information included in tables within these documents. This paper introduces a general-purpose system for linking entities to items in the Wikidata knowledge base. It describes how we adapt this system for linking domain-specific entities, especially for those entities embedded within tables drawn from COVID-19-related scientific literature. We describe the setup of an efficient offline instance of the system that enables our entity-linking approach to be more feasible in practice. As part of a broader approach to infer the semantic meaning of scientific tables, we leverage the structural and semantic characteristics of the tables to improve overall entity linking performance.

Long-term study on wastewater SARS-CoV-2 surveillance across United Arab Emirates.

Wastewater-based epidemiology (WBE) demonstrates an efficient tool to monitor and predict SARS-CoV-2 community distribution. Many countries across the world have adopted the technique, however, most of these studies were conducted for a short duration with a limited sampling size. In this study, long-term reliability and quantification of wastewater SARS-CoV-2 surveillance is reported via analyzing 16,858 samples collected from 453 different locations across the United Arab Emirates (UAE) from May 2020 to June 2022. The collected composite samples were first incubated at 60 °C followed by filtration, concentration, and then RNA extraction using commercially available kits. The extracted RNA was then analyzed by one-step RT-qPCR and RT-ddPCR, and the data was compared to the reported clinical cases. The average positivity rate in the wastewater samples was found to be 60.61 % (8.41-96.77 %), however, the positivity rate obtained from the RT-ddPCR was significantly higher than the RT-qPCR suggesting higher sensitivity of RT-ddPCR. Time-lagged correlation analysis indicated an increase in positive cases in the wastewater samples when the clinical positive cases declined suggesting that wastewater data are highly affected by the unreported asymptomatic, pre-symptomatic and recovering individuals. The weekly SARS-CoV-2 viral count in the wastewater samples are positively correlated with the diagnosed new clinical cases throughout the studied period and the studied locations. Viral count in wastewater peaked approximately one to two weeks prior to the peaks appearing in active clinical cases indicating that wastewater viral concentrations are effective in predicting clinical cases. Overall, this study further confirms the long-term sensitivity and robust approach of WBE to detect trends in SARS-CoV-2 spread and helps contribute to pandemic management.

Folding@home: Achievements from over 20 years of citizen science herald the exascale era.

Simulations of biomolecules have enormous potential to inform our understanding of biology but require extremely demanding calculations. For over 20 years, the Folding@home distributed computing project has pioneered a massively parallel approach to biomolecular simulation, harnessing the resources of citizen scientists across the globe. Here, we summarize the scientific and technical advances this perspective has enabled. As the project's name implies, the early years of Folding@home focused on driving advances in our understanding of protein folding by developing statistical methods for capturing long-timescale processes and facilitating insight into complex dynamical processes. Success laid a foundation for broadening the scope of Folding@home to address other functionally relevant conformational changes, such as receptor signaling, enzyme dynamics, and ligand binding. Continued algorithmic advances, hardware developments such as graphics processing unit (GPU)-based computing, and the growing scale of Folding@home have enabled the project to focus on new areas where massively parallel sampling can be impactful. While previous work sought to expand toward larger proteins with slower conformational changes, new work focuses on large-scale comparative studies of different protein sequences and chemical compounds to better understand biology and inform the development of small-molecule drugs. Progress on these fronts enabled the community to pivot quickly in response to the COVID-19 pandemic, expanding to become the world's first exascale computer and deploying this massive resource to provide insight into the inner workings of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus and aid the development of new antivirals. This success provides a glimpse of what is to come as exascale supercomputers come online and as Folding@home continues its work.

Folding@home: achievements from over twenty years of citizen science herald the exascale era (preprint)

Simulations of biomolecules have enormous potential to inform our understanding of biology but require extremely demanding calculations. For over twenty years, the Folding@home distributed computing project has pioneered a massively parallel approach to biomolecular simulation, harnessing the resources of citizen scientists across the globe. Here, we summarize the scientific and technical advances this perspective has enabled. As the project's name implies, the early years of Folding@home focused on driving advances in our understanding of protein folding by developing statistical methods for capturing long-timescale processes and facilitating insight into complex dynamical processes. Success laid a foundation for broadening the scope of Folding@home to address other functionally relevant conformational changes, such as receptor signaling, enzyme dynamics, and ligand binding. Continued algorithmic advances, hardware developments such as GPU-based computing, and the growing scale of Folding@home have enabled the project to focus on new areas where massively parallel sampling can be impactful. While previous work sought to expand toward larger proteins with slower conformational changes, new work focuses on large-scale comparative studies of different protein sequences and chemical compounds to better understand biology and inform the development of small molecule drugs. Progress on these fronts enabled the community to pivot quickly in response to the COVID-19 pandemic, expanding to become the world's first exascale computer and deploying this massive resource to provide insight into the inner workings of the SARS-CoV-2 virus and aid the development of new antivirals. This success provides a glimpse of what's to come as exascale supercomputers come online, and Folding@home continues its work.

Fear, Risk Perception, and Engagement in Preventive Behaviors for COVID-19 during Nationwide Lockdown in Nepal.

The world has faced huge negative effects from the COVID-19 pandemic between early 2020 and late 2021. Each country has implemented a range of preventive measures to minimize the risk during the COVID-19 pandemic. This study assessed the COVID-19-related fear, risk perception, and preventative behavior during the nationwide lockdown due to COVID-19 in Nepal. In a cross-sectional study, conducted in mid-2021 during the nationwide lockdown in Nepal, a total of 1484 individuals completed measures on fear of COVID-19, COVID-19 risk perception, and preventive behavior. A multiple linear regression analysis was used to identify factors associated with COVID-19 fear. The results revealed significant differences in the fear of COVID-19 in association with the perceived risk of COVID-19 and preventive behaviors. Age, risk perception, preventive behavior, and poor health status were significantly positively related to fear of COVID-19. Perceived risk and preventive behaviors uniquely predicted fear of COVID-19 over and above the effects of socio-demographic variables. Being female and unmarried were the significant factors associated with fear of COVID-19 among study respondents. Higher risk perception, poor health status, and being female were strong factors of increased fear of COVID-19. Targeted interventions are essential to integrate community-level mental health care for COVID-19 resilience.

Regenerative medicine applications: An overview of clinical trials

Insights into the use of cellular therapeutics, extracellular vesicles (EVs), and tissue engineering strategies for regenerative medicine applications are continually emerging with a focus on personalized, patient-specific treatments. Multiple pre-clinical and clinical trials have demonstrated the strong potential of cellular therapies, such as stem cells, immune cells, and EVs, to modulate inflammatory immune responses and promote neoangiogenic regeneration in diseased organs, damaged grafts, and inflammatory diseases, including COVID-19. Over 5,000 registered clinical trials on ClinicalTrials.gov involve stem cell therapies across various organs such as lung, kidney, heart, and liver, among other applications. A vast majority of stem cell clinical trials have been focused on these therapies' safety and effectiveness. Advances in our understanding of stem cell heterogeneity, dosage specificity, and ex vivo manipulation of stem cell activity have shed light on the potential benefits of cellular therapies and supported expansion into clinical indications such as optimizing organ preservation before transplantation. Standardization of manufacturing protocols of tissue-engineered grafts is a critical first step towards the ultimate goal of whole organ engineering. Although various challenges and uncertainties are present in applying cellular and tissue engineering therapies, these fields' prospect remains promising for customized patient-specific treatments. Here we will review novel regenerative medicine applications involving cellular therapies, EVs, and tissue-engineered constructs currently investigated in the clinic to mitigate diseases and possible use of cellular therapeutics for solid organ transplantation. We will discuss how these strategies may help advance the therapeutic potential of regenerative and transplant medicine.

Detection of SARS-CoV-2 in clinical and environmental samples using highly sensitive reduced graphene oxide (rGO)-based biosensor.

Quantitative RT-PCR (qRT-PCR) is the most commonly used diagnostic tool for SARS-CoV-2 detection during the COVID-19 pandemic. Despite its sensitivity and accuracy, qRT-PCR is a time-consuming method that requires expensive laboratories with highly trained personnel. In this work, on-site detection of SARS-CoV-2 in municipal wastewater was investigated for the first time. The wastewater was unprocessed and did not require any prefiltration, prior spiking with virus, or viral concentration in order to be suitable for use with the biosensor. The prototype reported here is a reduced graphene oxide (rGO)-based biosensor for rapid, sensitive and selective detection of SARS-CoV-2. The biosensor achieved a limit of detection (LOD) of 0.5 fg/mL in phosphate-buffered saline (PBS) and exhibited specificity when exposed to various analytes. The response time was measured to be around 240 ms. To further explore the capabilities of the biosensor in real clinical and municipal wastewater samples, three different tests were performed to determine the presence or absence of the virus: (i) qRT-PCR, (ii) a rapid antigen-based commercially available test (COVID-19 Test Strips), and (iii) the biosensor constructed and reported here. Taken together, our results demonstrate that a biosensor that can detect SARS-CoV-2 in clinical samples as well as unfiltered and unprocessed municipal wastewater is feasible.

Simulations of respiratory droplet spray

As part of the Summer High School Intensive in Next Generation Engineering (SHINE) with the University of Southern California's (USC) Viterbi School of Engineering, this work examines the transmission of the COVID-19 virus through respiratory droplets expelled from an infected individual. Social distancing and face masking protocols have been implemented to reduce the spread of droplets. This study aimed to assess the effectiveness of the recommendations using computational fluid dynamics simulations based upon the ANSYS Fluent Student Edition with two-dimensional axisymmetric simulations. The four most common ways of spreading respiratory droplets, including breathing, talking, coughing, and sneezing, were examined. Before completing the droplet spray simulations, several canonical jet flows were simulated to verify the validity of Fluent's application. Specifically, laminar and turbulent free jets were modeled and compared against experimental data. In addition, standard features of jets such as self-similarity, spreading ratios, and centerline velocity decay were verified from the solutions. Once the computations were validated, simulations were completed for each of the four cases at the six-foot recommended social distancing to determine the droplet concentration reduction. The simulations were run at increasing grid resolution to verify grid and time-step independence. Finally, the simulations were repeated for the case with face masks to assess the additional reduction of droplets reaching the receiver at the recommended distance. At the recommended social distancing with masking, the number of droplets coming into contact with others was reduced to negligible amounts. The simulations showed the recommended protocols are highly effective at reducing the transmission of COVID-19. © 2022, American Institute of Aeronautics and Astronautics Inc, AIAA. All rights reserved.

Optimization of an rGO-based biosensor for the sensitive detection of bovine serum albumin: Effect of electric field on detection capability.

Despite significant progress in the field of biosensing, the impact of electric field on biosensor detection capability and the feasibility of the biosensor application in wastewater has yet to be investigated. The objective of this study was to develop a low-cost, highly sensitive, and selective reduced graphene oxide (rGO)-based biosensor. The constructed biosensor consists of an in-house prepared GO and a four-terminal Kelvin sensing. Spin-coating was chosen as the deposition technique and results revealed an optimal GO number of layers and concentration of 7 and 2 mg/mL, respectively. Experiments to determine the effects of electric field on the performance of the biosensor showed significant changes in the biosensor surface, also presenting a direct impact on the biosensor functionality, such that the biosensor showed an increase in limit of detection (LOD) from 1 to 106 fg/mL when the applied voltage was increased from 0.0008 to 0.2 V. Furthermore, this study successfully explores a pilot scale setup, mimicking wastewater flow through sewage pipelines. The demonstrated improvements in the detection capability and sensitivity of this biosensor at optimized testing conditions make it a promising candidate for further development and deployment for the detection of protein analytes present at very low concentrations in aqueous solutions. In addition, the application of this biosensor could be extended to the detection of protein analytes of interest (such as the spike protein of SARS-CoV-2) in much more complex solutions, like wastewater.

Refined Capsid Structure of Human Adenovirus D26 at 3.4 Å Resolution.

Various adenoviruses are being used as viral vectors for the generation of vaccines against chronic and emerging diseases (e.g., AIDS, COVID-19). Here, we report the improved capsid structure for one of these vectors, human adenovirus D26 (HAdV-D26), at 3.4 Å resolution, by reprocessing the previous cryo-electron microscopy dataset and obtaining a refined model. In addition to overall improvements in the model, the highlights of the structure include (1) locating a segment of the processed peptide of VIII that was previously believed to be released from the mature virions, (2) reorientation of the helical appendage domain (APD) of IIIa situated underneath the vertex region relative to its counterpart observed in the cleavage defective (ts1) mutant of HAdV-C5 that resulted in the loss of interactions between the APD and hexon bases, and (3) the revised conformation of the cleaved N-terminal segments of pre-protein VI (pVIn), located in the hexon cavities, is highly conserved, with notable stacking interactions between the conserved His13 and Phe18 residues. Taken together, the improved model of HAdV-D26 capsid provides a better understanding of protein-protein interactions in HAdV capsids and facilitates the efforts to modify and/or design adenoviral vectors with altered properties. Last but not least, we provide some insights into clotting factors (e.g., FX and PF4) binding to AdV vectors.

Nanoparticle Engineered Photocatalytic Paints: A Roadmap to Self-Sterilizing against the Spread of Communicable Diseases

Applications of visible-light photocatalytic engineered nanomaterials in the preparation of smart paints are of recent origin. The authors have revealed a great potential of these new paints for self-sterilizing of the surfaces in hospitals and public places simply with visible light exposure and this is reported for the first time in this review. A recent example of a communicable disease such as COVID-19 is considered. With all precautions and preventions taken as suggested by the World Health Organization (WHO), COVID-19 has remained present for a longer time compared to other diseases. It has affected millions of people worldwide and the significant challenge remains of preventing infections due to SARS-CoV-2. The present review is focused on revealing the cause of this widespread disease and suggests a roadmap to control the spread of disease. It is understood that the transmission of SARS-CoV-2 virus takes place through contact surfaces such as doorknobs, packaging and handrails, which may be responsible for many preventable and nosocomial infections. In addition, due to the potent transmissibility of SARS-CoV-2, its ability to survive for longer periods on common touch surfaces is also an important reason for the spread of COVID-19. The existing antimicrobial cleaning technologies used in hospitals are not suitable, viable or economical to keep public places free from such infections. Hence, in this review, an innovative approach of coating surfaces in public places with visible-light photocatalytic nanocomposite paints has been suggested as a roadmap to self-sterilizing against the spread of communicable diseases. The formulations of different nanoparticle engineered photocatalytic paints with their ability to destroy pathogens using visible light, alongwith the field trials are also summarized and reported in this review. The potential suggestions for controlling the spread of communicable diseases are also listed at the end of the review.

Long-term SARS-CoV-2-specific and cross-reactive cellular immune responses correlate with humoral responses, disease severity, and symptomatology.

BACKGROUND: Cellular immune memory responses post coronavirus disease 2019 (COVID-19) have been difficult to assess due to the risks of contaminating the immune response readout with memory responses stemming from previous exposure to endemic coronaviruses. The work herein presents a large-scale long-term follow-up study investigating the correlation between symptomology and cellular immune responses four to five months post seroconversion based on a unique severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific peptide pool that contains no overlapping peptides with endemic human coronaviruses. METHODS: Peptide stimulated memory T cell responses were assessed with dual interferon-gamma (IFNγ) and interleukin (IL)-2 Fluorospot. Serological analyses were performed using a multiplex antigen bead array. RESULTS: Our work demonstrates that long-term SARS-CoV-2-specific memory T cell responses feature dual IFNγ and IL-2 responses, whereas cross-reactive memory T cell responses primarily generate IFNγ in response to SARS-CoV-2 peptide stimulation. T cell responses correlated to long-term humoral immune responses. Disease severity as well as specific COVID-19 symptoms correlated with the magnitude of the SARS-CoV-2-specific memory T cell response four to five months post seroconversion. CONCLUSION: Using a large cohort and a SARS-CoV-2-specific peptide pool we were able to substantiate that initial disease severity and symptoms correlate with the magnitude of the SARS-CoV-2-specific memory T cell responses.

Indian Academy of Cytologists National Guidelines for Cytopathology Laboratories for Handling Suspected and Positive COVID-19 (SARS-CoV-2) Patient Samples.

COVID-19, caused by the SARS-CoV-2 virus, has been declared a pandemic by the World Health Organization. This scenario has impacted the way we practice cytopathology. Cytology laboratories receive fresh and potentially infectious biological samples including those from the respiratory tract, from COVID-19 positive or suspected patients. Hence, the Indian Academy of Cytologists thought it necessary and fit to bring forth appropriate guidelines starting from transportation, receipt, processing, and reporting of samples in the COVID-19 era. The guidelines are prepared with the aim of safeguarding and protecting the health care personnel including laboratory staff, trainees and cytopathologists by minimizing exposure to COVID-19 so that they remain safe, in order to able to provide a continuous service. We hope that these national guidelines will be implemented across all cytopathology laboratories effectively.

Ex Vivo and In Vivo CD46 Receptor Utilization by Species D Human Adenovirus Serotype 26 (HAdV26).

Human adenovirus serotype 26 (Ad26) is used as a gene-based vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and HIV-1. However, its primary receptor portfolio remains controversial, potentially including sialic acid, coxsackie and adenovirus receptor (CAR), integrins, and CD46. We and others have shown that Ad26 can use CD46, but these observations were questioned on the basis of the inability to cocrystallize Ad26 fiber with CD46. Recent work demonstrated that Ad26 binds CD46 with its hexon protein rather than its fiber. We examined the functional consequences of Ad26 for infection in vitro and in vivo. Ectopic expression of human CD46 on Chinese hamster ovary cells increased Ad26 infection significantly. Deletion of the complement control protein domain CCP1 or CCP2 or the serine-threonine-proline (STP) region of CD46 reduced infection. Comparing wild-type and sialic acid-deficient CHO cells, we show that the usage of CD46 is independent of its sialylation status. Ad26 transduction was increased in CD46 transgenic mice after intramuscular (i.m.) injection but not after intranasal (i.n.) administration. Ad26 transduction was 10-fold lower than Ad5 transduction after intratumoral (i.t.) injection of CD46-expressing tumors. Ad26 transduction of liver was 1,000-fold lower than that ofAd5 after intravenous (i.v.) injection. These data demonstrate the use of CD46 by Ad26 in certain situations but also show that the receptor has little consequence by other routes of administration. Finally, i.v. injection of high doses of Ad26 into CD46 mice induced release of liver enzymes into the bloodstream and reduced white blood cell counts but did not induce thrombocytopenia. This suggests that Ad26 virions do not induce direct clotting side effects seen during coronavirus disease 2019 (COVID-19) vaccination with this serotype of adenovirus. IMPORTANCE The human species D Ad26 is being investigated as a low-seroprevalence vector for oncolytic virotherapy and gene-based vaccination against HIV-1 and SARS-CoV-2. However, there is debate in the literature about its tropism and receptor utilization, which directly influence its efficiency for certain applications. This work was aimed at determining which receptor(s) this virus uses for infection and its role in virus biology, vaccine efficacy, and, importantly, vaccine safety.

Epitope-Analyzer: A structure-based webtool to analyze broadly neutralizing epitopes.

The antigenic epitope regions of pathogens (e.g., viruses) are recognized by antibodies (Abs) and subsequently cleared by the host immune system, thereby protecting us from disease. Some of these epitopes are conserved among different variants or subgroups of pathogens (e.g., Influenza (FLU) viruses, Coronaviruses), hence can be targeted for potential broad-neutralization. Here we report a web-based tool, Epitope Analyzer (EA), that rapidly identifies conformational epitope and paratope residues in an antigen-antibody complex structure. Furthermore, the tool provides the ways and means to analyze broadly neutralizing epitopes by comparing the equivalent epitope residues in similar antigen structures. The similarity in the epitope residues between (multiple) pairs of similar antigen molecules suggest the presence of conserved epitopes that can be targeted by broadly neutralizing antibodies. These details can be used as a guide in developing effective treatments, such as the design of novel vaccines and formulation of cocktail of broadly neutralizing antibodies, against multiple variants or subgroups of viruses. The web application can be freely accessed from the URL, http://viperdb.scripps.edu/ea.php.

Recent advances in the biosensors application for the detection of bacteria and viruses in wastewater.

The presence of disease-causing pathogens in wastewater can provide an excellent diagnostic tool for infectious diseases. Biosensors are far superior to conventional methods used for regular infection screening and surveillance testing. They are rapid, sensitive, inexpensive portable and carry no risk of exposure in their detection schemes. In this context, this review summarizes the most recently developed biosensors for the detection of bacteria and viruses in wastewater. The review also provides information on the new detection methods aimed at screening for SARS-CoV-2, which has now caused more than 4 million deaths. In addition, the review highlights the potential behind on-line and real-time detection of pathogens in wastewater pipelines. Most of the biosensors reported were not targeted to wastewater samples due to the complexity of the matrix. However, this review highlights on the performance factors of recently developed biosensors and discusses the importance of nanotechnology in amplifying the output signals, which in turn increases the accuracy and reliability of biosensors. Current research on the applicability of biosensors in wastewater promises a dramatic change to the conventional approach in the field of medical screening.

Coronavirus disease 2019 convalescent children: outcomes after congenital heart surgery.

BACKGROUND: Children with exposure to coronavirus disease 2019 in recent times (asymptomatic or symptomatic infection) approaching congenital heart surgery programme are in increasing numbers. Understanding outcomes of such children will help risk-stratify and guide optimisation prior to congenital heart surgery. OBJECTIVE: The objective of the present study was to determine whether convalescent coronavirus disease 2019 children undergoing congenital heart surgery have any worse mortality or post-operative outcomes. DESIGN: Consecutive children undergoing congenital heart surgery from Oct 2020 to May 2021 were enrolled after testing for reverse transcription-polymerase chain reaction or rapid antigen test and immunoglobulin G antibody prior to surgery. Convalescent coronavirus disease 2019 was defined in any asymptomatic patient positive for immunoglobulin G antibodies and negative for reverse transcription-polymerase chain reaction or rapid antigen test anytime 6 weeks prior to surgery. Control patients were negative for any of the three tests. Mortality and post-operative outcomes were compared among the groups. RESULTS: One thousand one hundred and twenty-nine consecutive congenital heart surgeries were stratified as convalescence and control. Coronavirus disease 2019 Convalescent (n = 349) and coronavirus disease 2019 control (n = 780) groups were comparable for all demographic and clinical factors except younger and smaller kids in control. Convalescent children had no higher mortality, ventilation duration, ICU and hospital stay, no higher support with extracorporeal membrane oxygenation, high flow nasal cannula, no higher need for re-intubations, re-admissions, and no higher infections as central line-associated bloodstream infection, sternal site infection, and ventilator-associated pneumonia on comparison with coronavirus disease 2019 control children. CONCLUSIONS: Convalescent coronavirus disease 2019 does not have any unfavourable outcomes as compared to coronavirus disease 2019 control children. Positive immunoglobulin G antibody screening prior to surgery is suggestive of convalescence and supports comparable outcomes on par with control peers.

Comparative analysis of COVID-19 case fatality rate between two waves in Nepal.

The first COVID-19 case in Nepal was reported on January 23, 2020. Then infection, then, started to spread gradually, and October marked the most devastating increase in COVID-19 cases of the year 2020. Compared with the October 2020 peak in Nepal, the May 2021 peak of COVID-19 observed 2- and 10-fold rise in new cases and deaths per day, respectively. Given that this surprising increase in the death rate was not observed in other countries, this study analyzed the COVID-19 case fatality rates between the two peaks in Nepal. We found an increase in death rates among younger adults and people without comorbidities.