SARS-Cov-2 Spike-S1 Antigen Test Strip with High Sensitivity Endowed by High-Affinity Antibodies and Brightly Fluorescent QDs/Silica Nanospheres.

The extensive research into developing novel strategies for detecting respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens in clinical specimens, especially the sensitive point-of-care testing method, is still urgently needed to reach rapid screening of viral infections. Herein, a new lateral flow immunoassay (LFIA) platform was reported for the detection of SARS-CoV-2 spike-S1 protein antigens, in which four sensitive and specific SARS-CoV-2 mouse monoclonal antibodies (MmAbs) were tailored by using quantum dot (QD)-loaded dendritic mesoporous silica nanoparticles modified further for achieving the -COOH group surface coating (named Q/S-COOH nanospheres). Importantly, compact QD adsorption was achieved in mesoporous channels of silica nanoparticles on account of highly accessible central-radial pores and electrostatic interactions, leading to significant signal amplification. As such, a limit of detection for SARS-CoV-2 spike-S1 testing was found to be 0.03 ng/mL, which is lower compared with those of AuNPs-LFIA (traditional colloidal gold nanoparticles, Au NPs) and enzyme-linked immunosorbent assay methods. These results show that optimizing the affinity of antibody and the intensity of fluorescent nanospheres simultaneously is of great significance to improve the sensitivity of LFIA.

Immune response to COVID-19 vaccination in a population with and without a previous SARS-CoV-2 infection.

PURPOSE: To evaluate IgG production in a group of vaccinated and unvaccinated subjects previously infected, or not, with SARS-CoV-2. METHODS: A total of 316 subjects were enrolled at different times after vaccination and/or infection. IgG against target S1 subunit of the spike protein of SARS-COV-2 was assessed by a chemiluminescent microparticle immunoassay. Participant data was collected using a clinical-epidemiological survey. RESULTS: A total of 56.2% (n = 146) of our cohort was vaccinated, with 27.5% (n = 36) reporting a previous infection. Of these, all were IgG positive at the time of the study, regardless of gender, age category, vaccine type, and elapsed time since vaccination. The vaccinated group without a previous infection (72.5%, n = 95) showed a slightly lower IgG seropositivity and median values, overall, although significantly higher in females and lower with the ChAdOx1 nCoV-19 (AstraZeneca) vaccine. Vaccinated subjects above the age of 65 showed a trend towards higher median IgG values (13,911.0 AU/mL), when previously infected with SARS-CoV-2, but comparatively lower IgG median value (5158.7 AU/mL) in its absence. In all vaccinated groups, IgG antibody production increased at 1-2 weeks, peaking at 4-6 weeks. Afterward, IgG decreased progressively but almost all subjects (97.7%, n = 128) were seropositive for the remainder of our study. Fully vaccinated individuals with a past infection showed a lower IgG rate of decrease versus their uninfected counterparts (17.9 vs 22.6%, respectively). CONCLUSION: Our findings suggest a higher effect of vaccination on the production IgG antibodies, as opposed to natural infection. Nonetheless, in general, antibody titers waned rapidly.

Machine learning for optimizing daily COVID-19 vaccine dissemination to combat the pandemic.

Introduction: Vaccines are the most important instrument for bringing the pandemic to a close and saving lives and helping to reduce the risks of infection. It is important that everyone has equal access to immunizations that are both safe and effective. There is no one who is safe until everyone gets vaccinated. COVID-19 vaccinations are a game-changer in the fight against diseases. In addition to examining attitudes toward these vaccines in Africa, Asia, Oceania, Europe, North America, and South America, the purpose of this paper is to predict the acceptability of COVID-19 vaccines and study their predictors. Materials and methods: Kaggle datasets are used to estimate the prediction outcomes of the daily COVID-19 vaccination to prevent a pandemic. The Kaggle data sets are classified into training and testing datasets. The training dataset is comprised of COVID-19 daily data from the 13th of December 2020 to the 13th of June 2021, while the testing dataset is comprised of COVID-19 daily data from the 14th of June 2021 to the 14th of October 2021. For the prediction of daily COVID-19 vaccination, four well-known machine learning algorithms were described and used in this study: CUBIST, Gaussian Process (GAUSS), Elastic Net (ENET), Spikes, and Slab (SPIKES). Results: Among the models considered in this paper, CUBIST has the best prediction accuracy in terms of Mean Absolute Scaled Error (MASE) of 9.7368 for Asia, 2.8901 for America, 13.2169 for Oceania, and 3.9510 for South America respectively. Conclusion: This research shows that machine learning can be of great benefit for optimizing daily immunization of citizens across the globe. And if used properly, it can help decision makers and health administrators to comprehend immunization rates and create strategies to enhance them.

Simple neutralization test report: Do probiotics contribute to COVID-19 therapy?

Background: There is an urgent need to identify effective therapy to treat coronavirus diseases 2019 (COVID-19). Supplement consumption is becoming popular in this pandemic era. An example of this is probiotic consumption to improve the host's immune system. Objective: This study aimed to prove whether antibodies from people taking probiotics could influence lactate dehydrogenase (LDH), adenosine triphosphate (ATP) values, and cell viability in vitro in peripheral blood mononuclear cells (PBMCs) inoculated with the SARS-CoV-2 spike protein as COVID-19 cells models. Methods: This was an experimental study with control and intervention groups, totally in 12 groups divided based on antibody levels, probiotic intervention, probiotic non-intervention group, SARS-CoV-2 infection group, and non-SARS-CoV-2 infection group. In vitro assays were carried out on PBMC cell cultures inoculated with S1 SARS-CoV-2 recombinant as a COVID-19 cell model. The COVID-19 cell model was given antibodies divided into three antibody level groups: sRBD levels of <3, 325.76 and 646.18. The cytotoxicity assessment examined increased levels of LDH, cytopathic activity by measuring ATP levels, and cell viability by XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) assay. Data were analyzed with SPSS 21 for Windows. Results: This study showed a significant difference in the LDH value (p < 0.001) between each group. The difference in ATP values between groups was significant (p < 0.001). Meanwhile, the cell viability examination found that there was a tendency of decreased XTT (cell viability in %) when there was an increase of LDH and ATP. Conclusion: The change of LDH values occurred most in the antibody group that did not consume probiotics. The highest cytopathic activity based on the ATP values occurred in the infected cell culture group with antibody levels of 325.76 and consuming probiotics. In addition, LDH and ATP activities provided evidence of a significant influence on cell viability.

An Optical Modeling Framework for Coronavirus Detection Using Graphene-Based Nanosensor.

The outbreak of the COVID-19 virus has faced the world with a new and dangerous challenge due to its contagious nature. Hence, developing sensory technologies to detect the coronavirus rapidly can provide a favorable condition for pandemic control of dangerous diseases. In between, because of the nanoscale size of this virus, there is a need for a good understanding of its optical behavior, which can give an extraordinary insight into the more efficient design of sensory devices. For the first time, this paper presents an optical modeling framework for a COVID-19 particle in the blood and extracts its optical characteristics based on numerical computations. To this end, a theoretical foundation of a COVID-19 particle is proposed based on the most recent experimental results available in the literature to simulate the optical behavior of the coronavirus under varying physical conditions. In order to obtain the optical properties of the COVID-19 model, the light reflectance by the structure is then simulated for different geometrical sizes, including the diameter of the COVID-19 particle and the size of the spikes surrounding it. It is found that the reflectance spectra are very sensitive to geometric changes of the coronavirus. Furthermore, the density of COVID-19 particles is investigated when the light is incident on different sides of the sample. Following this, we propose a nanosensor based on graphene, silicon, and gold nanodisks and demonstrate the functionality of the designed devices for detecting COVID-19 particles inside the blood samples. Indeed, the presented nanosensor design can be promoted as a practical procedure for creating nanoelectronic kits and wearable devices with considerable potential for fast virus detection.

SARS-CoV-2 Infection-Of Music and Mechanics of Its Spikes! A Perspective.

The COVID-19 pandemic has been inflicted upon humanity by the SARS-CoV-2 virus, the latest insidious incarnation of the coronaviruses group. While in its wake intense scientific research has produced breakthrough vaccines and cures, there still exists an immediate need to further understand the origin, mechanobiology and biochemistry, and destiny of this virus so that future pandemics arising from similar coronaviruses may be contained more effectively. In this Perspective, we discuss the various evidential findings of virus propagation and connect them to respective underpinning cellular biomechanical states leading to corresponding manifestations of the viral activity. We further propose avenues to tackle the virus, including from a "musical" vantage point, and contain its relentless strides that are currently afflicting the global populace.

Quantitative measurement of IgG to SARS-CoV-2 antigens using monoclonal antibody-based enzyme-linked immunosorbent assays.

OBJECTIVE: Standardised quantitative analysis of the humoral immune response to SARS-CoV-2 antigens may be useful for estimating the extent and duration of immunity. The aim was to develop enzyme-linked immunosorbent assays (ELISAs) for the quantification of human IgG antibodies against SARS-CoV-2 antigens. METHODS: Enzyme-linked immunosorbent assays were developed based on monoclonal antibodies against human IgG and recombinant SARS-CoV-2 antigens (Spike-S1 and Nucleocapsid). The WHO 67/086 immunoglobulin and WHO 20/136 SARS-CoV-2 references were used for standardisation. Sera of a study group of COVID-19-positive subjects (n = 144), pre-pandemic controls (n = 135) and individuals vaccinated with BioNTech-Pfizer BNT162b2 vaccine (n = 48) were analysed. The study group sera were also tested using EuroImmun SARS-CoV-2-ELISAs and a quantitative S1-specific fluorescence enzyme immunoassay (FEIA) from Thermo Fisher. RESULTS: The ELISA results were repeatable and traceable to international units because of their parallelism to both WHO references. In the study group, median anti-S1-IgG concentrations were 102 BAU mL-1, compared to 100 and 1457 BAU mL-1 in the vaccination group after first and second vaccination, respectively. The ELISAs achieved an area under the curve (AUC) of 0.965 (S1) and 0.955 (Nucleocapsid) in receiver operating characteristic (ROC) analysis, and a specificity of 1 (S1) and 0.963 (Nucleocapsid) and sensitivity of 0.903 (S1) and 0.833 (Nucleocapsid) at the maximum Youden index. In comparison, the commercial assays (S1-FEIA, S1 and Nucleocapsid ELISA EuroImmun) achieved sensitivities of 0.764, 0.875 and 0.882 in the study group, respectively. CONCLUSIONS: The quantitative ELISAs to measure IgG binding to SARS-CoV-2 antigens have good analytical and clinical performance characteristics and units traceable to international standards.

Variable levels of spike and ORF1ab RNA in post-mortem lung samples of SARS-CoV-2-positive subjects: comparison between ISH and RT-PCR.

Post-mortem examination plays a pivotal role in understanding the pathobiology of the SARS-CoV-2; thus, the optimization of virus detection on the post-mortem formalin-fixed paraffin-embedded (FFPE) tissue is needed. Different techniques are available for the identification of the SARS-CoV-2, including reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), in situ hybridization (ISH), and electron microscopy. The main goal of this study is to compare ISH versus RT-PCR to detect SARS-CoV-2 on post-mortem lung samples of positive deceased subjects. A total of 27 samples were analyzed by RT-PCR targeting different viral RNA sequences of SARS-CoV-2, including envelope (E), nucleocapsid (N), spike (S), and open reading frame (ORF1ab) genes and ISH targeting S and Orf1ab. All 27 cases showed the N gene amplification, 22 out of 27 the E gene amplification, 26 out of 27 the S gene amplification, and only 6 the ORF1ab gene amplification. The S ISH was positive only in 12 out of 26 cases positive by RT-PCR. The S ISH positive cases with strong and diffuse staining showed a correlation with low values of the number of the amplification cycles by S RT-PCR suggesting that ISH is a sensitive assay mainly in cases carrying high levels of S RNA. In conclusion, our findings demonstrated that ISH assay has lower sensitivity to detect SARS-CoV-2 in FFPE compared to RT-PCR; however, it is able to localize the virus in the cellular context since it preserves the morphology.

Telesimulation Innovation on the Teaching of SPIKES Model on Sharing Bad News.

Coronavirus disease 2019 (COVID-19) brought to the world, an unprecedented emergency, which dramatically affected the face-to-face teaching in higher education academia. University faculty and students had to shift overnight to an online and remote course instruction. They were neither trained nor prepared and had limited resources and infrastructure. Palliative Care and Oncology Stream Faculty at Aga Khan University, School of Nursing and Midwifery, Pakistan, piloted an innovative academic project using telesimulation (TS). Trainee nurse interns were taught communication skills and the art of breaking bad news to palliative clients using the SPIKES model through TS. To incorporate best practices for simulation-based experiences, we used the International Nursing Association for Clinical Simulation and Learning to standardize and implement TS with 141 interns. This review article documents how the faculty planned and implemented the TS strategy during COVID-19. It outlines the challenges and the lessons learnt from implementation and feedback from faculty and students. This information could be useful in the future execution of TS, in any communication and counseling course, since COVID-19 has impacted the future educational course design and pedagogy worldwide.

Molecular Level Dissection of Critical Spike Mutations in SARS-CoV-2 Variants of Concern (VOCs): A Simplified Review.

SARS-CoV-2 virus during its spread in the last one and half year has picked up critical changes in its genetic code i.e. mutations, which have leads to deleterious epidemiological characteristics. Due to critical role of spike protein in cell entry and pathogenesis, mutations in spike regions have been reported to enhance transmissibility, disease severity, possible escape from vaccine-induced immune response and reduced diagnostic sensitivity/specificity. Considering the structure-function impact of mutations, understanding the molecular details of these key mutations of newly emerged variants/lineages is of urgent concern. In this review, we have explored the literature on key spike mutations harbored by alpha, beta, gamma and delta 'variants of concern' (VOCs) and discussed their molecular consequences in the context of resultant virus biology. Commonly all these VOCs i.e. B.1.1.7, B.1.351, P.1 and B.1.617.2 lineages have decisive mutation in Receptor Binding Motif (RBM) region and/or region around Furin cleavage site (FCS) of spike protein. In general, mutation induced disruption of intra-molecular interaction enhances molecular flexibility leading to exposure of spike protein surface in these lineages to make it accessible for inter-molecular interaction with hACE2. A disruption of spike antigen-antibody inter-molecular interactions in epitope region due to the chemical nature of substituting amino acid hampers the neutralization efficacy. Simplified surveillance of mutation induced changes and their consequences at molecular level can contribute in rationalizing mutation's impact on virus biology. It is believed that molecular level dissection of these key spike mutation will assist the future investigations for a more resilient outcome against severity of COVID-19.

Exploring SARS-CoV-2 Spikes Glycoproteins for Designing Potential Antiviral Targets.

Till today, the globe is still struggling with the newly emerging infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and known as coronavirus disease 2019 (COVID-19). It has resulted in multiple fatalities from SARSs all around the world. A year after the global pandemic, the World Health Organization (WHO) has reported more than 79 million confirmed cases of COVID-19 and over 1.7 million deaths, making it one of the worst and most difficult pandemics encompassed in the modern history. The ongoing triad of escalating infections, mortality, and economic loss has urgently called for recognizing SARS-CoV-2 cell entry mechanisms as a crucial step in the initial stages of infection and to which possible interventional strategies should be targeted. To mediate host cell infections, Coronaviruses utilize the immunogenic studded spikes glycoproteins on its surface as a key factor for attachment, fusion, and entrance to host cells. Herein, we shed the light on a potential strategy involving disruption of SARS-CoV-2 S protein interaction with host cell receptors through design of neutralizing antibodies targeting receptor binding domain in S1 subunit, small peptide inhibitors, peptide fusion inhibitors against S2, host cell angiotensin converting enzymes 2 (ACE2), and protease inhibitors, aiming to pave the way for controlling viral cell entrance. In this review, we also highlight the recent research advances in the antiviral drugs that target the highly exposed spike protein, aiming to stem the COVID-19 pandemic.

Communicating bad news in the context of COVID-19.

Communicating bad news is one of the most frequent activities in hospitals, for which some recommendations have been adapted to the needs within the coronavirus-2 disease (COVID-19) context. This document presents nine steps to deliver bad news (face to face or remotely) adapted to the COVID-19 context from two international protocols (SPIKES and GRIEV_ING). The importance of promoting physical and emotional self-care skills in health personnel is also described, as well as psychological first aid strategies to address the emotional response of the family member who receives the news. Finally, the limitations and advantages of the proposal should be considered.

La comunicación de malas noticias es una de las actividades más frecuentes en los hospitales dentro del contexto de la COVID-19. A pesar de su alta frecuencia, existen pocas recomendaciones adaptadas a las necesidades que el contexto de la COVID-19 demanda. Debido a lo anterior, en el presente escrito se presentan nueve pasos para dar malas noticias (cara a cara o por vía remota) de dos protocolos internacionales (SPIKES y GRIEV_ING) adaptados a las necesidades de transmisión de información de la COVID-19. Se describe también la importancia de promover habilidades de autocuidado físico y emocional en el personal de salud, así como estrategias de primeros auxilios psicológicos para el abordaje de la respuesta emocional del familiar que recibe la noticia. Finalmente, se deben considerar las limitaciones y ventajas de la propuesta.

Why Are Viruses Spiked?

Many viruses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and human immunodeficiency virus (HIV), have a structure consisting of spikes protruding from an underlying spherical surface. Research in biological and colloidal sciences has revealed secrets of why spikes exist on virus surfaces. Specifically, the spikes favor virus attachment on surfaces via receptor-specific interactions (RSIs), mediate the membrane fusion, and determine or change viral tropism. The spikes also facilitate viruses to approach surfaces before attachment and subsequently escape back to the environment if RSIs do not occur (i.e., easy come and easy go). Therefore, virus spikes create the paradox of having a large capacity for binding with cells (high infectivity) and meanwhile great mobility in the environment. Such structure-function relationships have important implications for the fabrication of virus-like particles and analogous colloids (e.g., hedgehog- and raspberry-like particles) for applications such as the development of antiviral vaccines and drug delivery.

Effect of receptors on the resonant and transient harmonic vibrations of Coronavirus.

The paper is concerned with the vibration characteristics of the Coronavirus family. There are some 25-100 receptors, commonly called spikes protruding from the envelope shell of the virus. Spikes, resembling the shape of a hot air balloon, may have a total mass similar to the mass of the lipid bi-layer shell. The lipid proteins of the virus are treated as homogeneous elastic material and the problem is formulated as the interaction of thin elastic shell with discrete masses, modeled as short conical cross-sectional beams. The system is subjected to ultrasonic excitation. Using the methods of structural acoustics, it is shown that the scattered pressure is very small and the pressure on the viral shell is simply the incident pressure. The modal analysis is performed for a bare shell, a single spike, and the spike-decorated shell. The predicted vibration frequencies and modes are shown to compare well with the newly derived closed-form solutions for a single spike and existing analytical solutions for thin shells. The fully nonlinear dynamic simulation of the transient response revealed the true character of the complex interaction between local vibration of spikes and global vibration of the multi-degree-of-freedom system. It was shown that harmonic vibration at or below the lowest resonant modes can excite large amplitude vibration of spikes. The associated maximum principal strain in a spike can reach large values in a fraction of a millisecond. Implications for possible tearing off spikes from the shell are discussed. Another important result is that after a finite number of cycles, the shell buckles and collapses, developing internal contacts and folds with large curvatures and strains exceeding 10%. For the geometry and elastic properties of the SARS-CoV-2 virus, these effects are present in the range of frequencies close to the ones used for medical ultrasound diagnostics.

Host Receptors of Influenza Viruses and Coronaviruses-Molecular Mechanisms of Recognition.

Among the four genera of influenza viruses (IVs) and the four genera of coronaviruses (CoVs), zoonotic αIV and ßCoV have occasionally caused airborne epidemic outbreaks in humans, who are immunologically naïve, and the outbreaks have resulted in high fatality rates as well as social and economic disruption and losses. The most devasting influenza A virus (IAV) in αIV, pandemic H1N1 in 1918, which caused at least 40 million deaths from about 500 million cases of infection, was the first recorded emergence of IAVs in humans. Usually, a novel human-adapted virus replaces the preexisting human-adapted virus. Interestingly, two IAV subtypes, A/H3N2/1968 and A/H1N1/2009 variants, and two lineages of influenza B viruses (IBV) in ßIV, B/Yamagata and B/Victoria lineage-like viruses, remain seasonally detectable in humans. Both influenza C viruses (ICVs) in γIV and four human CoVs, HCoV-229E and HCoV-NL63 in αCoV and HCoV-OC43 and HCoV-HKU1 in ßCoV, usually cause mild respiratory infections. Much attention has been given to CoVs since the global epidemic outbreaks of ßSARS-CoV in 2002-2004 and ßMERS-CoV from 2012 to present. ßSARS-CoV-2, which is causing the ongoing COVID-19 pandemic that has resulted in 890,392 deaths from about 27 million cases of infection as of 8 September 2020, has provoked worldwide investigations of CoVs. With the aim of developing efficient strategies for controlling virus outbreaks and recurrences of seasonal virus variants, here we overview the structures, diversities, host ranges and host receptors of all IVs and CoVs and critically review current knowledge of receptor binding specificity of spike glycoproteins, which mediates infection, of IVs and of zoonotic, pandemic and seasonal CoVs.

Dynamics of epidemics: Impact of easing restrictions and control of infection spread.

During an infectious disease outbreak, mathematical models and computational simulations are essential tools to characterize the epidemic dynamics and aid in design public health policies. Using these tools, we provide an overview of the possible scenarios for the COVID-19 pandemic in the phase of easing restrictions used to reopen the economy and society. To investigate the dynamics of this outbreak, we consider a deterministic compartmental model (SEIR model) with an additional parameter to simulate the restrictions. In general, as a consequence of easing restrictions, we obtain scenarios characterized by high spikes of infections indicating significant acceleration of the spreading disease. Finally, we show how such undesirable scenarios could be avoided by a control strategy of successive partial easing restrictions, namely, we tailor a successive sequence of the additional parameter to prevent spikes in phases of low rate of transmissibility.

The dual impact of ACE2 in COVID-19 and ironical actions in geriatrics and pediatrics with possible therapeutic solutions.

The novel corona virus disease has shaken the entire world with its deadly effects and rapid transmission rates, posing a significant challenge to the healthcare authorities to develop suitable therapeutic solution to save lives on earth. The review aims to grab the attention of the researchers all over the globe, towards the role of ACE2 in COVID-19 disease. ACE2 serves as a molecular target for the SARS-CoV-2, to enter the target cell, by interacting with the viral glycoprotein spikes. However, the complexity began when numerous studies identified the protective response of ACE2 in abbreviating the harmful effects of vasoconstrictor, anti-inflammatory peptide, angiotensin 2, by mediating its conversion to angiotensin-(1-7), which exercised antagonistic actions to angiotensin 2. Furthermore, certain investigations revealed greater resistance among children as compared to the geriatrics, towards COVID-19 infection, despite the elevated expression of ACE2 in pediatric population. Based upon such evidences, the review demonstrated possible therapeutic interventions, targeting both the protective and deleterious effects of ACE2 in COVID-19 disease, primarily inhibiting ACE2-virus interactions or administering soluble ACE2. Thus, the authors aim to provide an opportunity for the researchers to consider RAAS system to be a significant element in development of suitable treatment regime for COVID-19 pandemic.