Boosting the Immune System with Vitamin D: Special Focus on Prevention of COVID-19 and Complications

In addition to the classical functions of the musculoskeletal system and calcium homeostasis, the function of vitamin D as an immune modulator is well established. The vitamin D receptors and enzymes that metabolize vitamin D are ubiquitously expressed in most cells in the body, including T and B lymphocytes, antigen-presenting cells, monocytes, macrophages and natural killer cells that trigger immune and antimicrobial responses. Many in vitro and in vivo studies revealed that vitamin D promotes tolerogenic immunological action and immune modulation. Vitamin D adequacy positively influences the expression and release of antimicrobial peptides, such as cathelicidin, defensin, and anti-inflammatory cytokines, and reduces the expression of proinflammatory cytokines. Evidence suggestss that vitamin D's protective immunogenic actions reduce the risk, complications, and death from COVID-19. On the contrary, vitamin D deficiency worsened the clinical outcomes of viral respiratory diseases and the COVID-19-related cytokine storm, acute respiratory distress syndrome, and death. The study revealed the need for more preclinical studies and focused on well-designed clinical trials with adequate sizes to understand the role of vitamin D on the pathophysiology of immune disorders and mechanisms of subduing microbial infections, including COVID-19.Copyright © 2023 Bentham Science Publishers.

Animal Pre-Clinical Study of an Inactivated SARS-CoV-2 Vaccine candidate (Osvid-19 ): Immunogenicity, Protective, and Safety Aspects

Background: This study aimed to evaluate the outcomes of preclinical studies on the safety and immunogenicity of an inactivated COVID-19 vaccine candidate to warrant further clinical evaluation. Method(s): SARS-CoV-2 positive nasopharyngeal swab specimens were confirmed by real-time polymerase chain reaction and next-generation sequencing. The safety and immunogenicity tests of the COVID-19 vaccine were carried out in rats and Rhesus monkeys, and Balb/C mice and Rhesus monkeys, respectively. Result(s): The candidate vaccine was well tolerated and induced promising levels of SARS-CoV-2- specific IgG1, IgG2a, and Granzyme B in Balb/C mice, and anti-SARS-CoV-2 spike IgG and neutralizing antibodies in Rhesus monkeys. Based on cVNT results, the inactivated vaccine in 0.5 and 1 microg/100 microL doses was able to induce a neutralizing effect against the SARS-CoV-2 virus up to a dilution of 1:512 and 1:1000. The protective efficacy of the vaccine candidate was challenged with 2 x108 PFU of live viruses and confirmed by lung CT scan and histopathological evaluations compared to the control group. Repeated intramuscular injection of the candidate vaccine was generally well-tolerated in Rats and Rhesuses. No significant side effects were observed in rats injected with ten full human doses and in the Rhesus monkeys with three full human doses. Conclusion(s): Based on the findings presented in this study, it is recommended that this vaccine be moved into human testing commencing with a phase I clinical trial.Copyright © 2021 Muslim OT et al.

2022 CAS Virtual Annual Meeting

The proceedings contain 63 papers. The topics discussed include: a retrospective study to optimize post-anesthetic recovery time after ambulatory lower limb orthopedic procedures at a tertiary care hospital in Canada;a virtual airway evaluation as good as the real thing?;airway management during in hospital cardiac arrest by a consultant led airway management team during the COVID-19 pandemic: a prospective and retrospective quality assurance project;prevention of cautery induced airway fire using saline filled endotracheal tube cuffs: a study in a trachea airway fire model;smart phone assisted retrograde illumination versus conventional laryngoscope illumination for orotracheal intubation: a prospective comparative trial;time to single lung isolation in massive pulmonary hemorrhage simulation using a novel bronchial blocker and traditional techniques;cannabinoid type 2 receptor activation ameliorates acute lung injury induced systemic inflammation;bleeding in patients with end-stage liver disease undergoing liver transplantation and fibrinogen level: a cohort study;endovascular Vena Cavae occlusion in right anterior mini-thoracoscopic approach for tricuspid valve in patients with previous cardiac surgery;and mesenchymal stem cell extracellular vesicles as a novel, regenerative nanotherapeutic for myocardial infarction: a preclinical systematic review.

Scientific debrief: cirrhosis

Globally, hepatitis C (26%), alcohol (24%), and hepatitis B (23%) contribute almost equally to the global burden of cirrhosis. The contribution from nonalcoholic fatty liver disease (8%) is small but increasing. Patients with acutely decompensated cirrhosis have a dismal prognosis and frequently progress to acuteon-chronic liver failure, which is characterised by hepatic and extrahepatic organ failure, Cardiovascular alterations including portal hypertension trigger the formation of portocaval shunts and varices. Systemic under filling and arterial hypotension is compensated by vasoconstriction but might decline into a state of aggravated portal hypertension and cirrhotic cardiomyopathy, leading to a hyperdynamic state, microvascular dysfunction and reduced organ perfusion culminating in decompensation. The immune system is dysfunctional showing a contrary co-existence of immune paralysis and immune overstimulation leading to secondary infections and inflammatory response syndrome aggravating cardiovascular alterations but also initiating tissue injury and metabolic alteration. This transition from compensated to decompensated cirrhosis is characterised by the occurrence of ascites, variceal bleeding and/or hepatic encephalopathy or organ failures (in the case of ACLF. Precipitating events for ACLF vary between Western countries (bacterial infection, alcohol intake) and Eastern countries (flare of HBV, superimposed HAV or HEV). In the majority of patients, systemic inflammation is a major driver of progression from compensated to decompensated cirrhosis. Once the first episode of AD develops, systemic inflammation follows a chronic course, with transient periods of aggravation due to proinflammatory precipitants or bursts of bacterial translocation resulting in repeated episodes of AD. The multistate model describing the clinical outcomes of decompensated cirrhosis has been well validated. State 3 is defined by the occurrence of variceal bleeding alone, state 4 by any single non-bleeding event, state 5 by any 2 or more events and the late decompensate state by any event with organ failures either with or without ACLF. 5-year mortality across states from 3 to 5 is in the order of, respectively: 20%, 30%, 88%. With late decompensation mortality ranges between 60 and 80% at 1 year. Cirrhosis is increasingly common and morbid. Optimal utilisation of therapeutic strategies to prevent and control the complications of cirrhosis are central to improving clinical and patient-reported outcomes. Aetiology-focused therapies that can prevent cirrhosis and its complications. These include anti-viral therapies, psychopharmacological therapy for alcohol-use disorder, management of hepatic encephalopathy (HE), ascites, hepatorenal syndrome, non-pain symptoms of cirrhosis including pruritis, muscle cramps, sexual dysfunction and fatigue, and reduce the risk of hepatocellular carcinoma. New disease-modifying agents are expected to be identified in the next few years by systematic drug repurposing and the development of novel molecules currently undergoing pre-clinical or early clinical testing. COVID-19 continues to pose a significant healthcare challenge throughout the world. Comorbidities including diabetes and hypertension are associated with a significantly higher mortality risk. Cirrhosis is associated with an increased risk of all-cause mortality in COVID-19 infection compared to non-cirrhotic patients. Patients with cirrhosis should be considered for targeted public health interventions to prevent COVID-19 infection, such as shielding and prioritisation of vaccination.

Role of toll-like receptors in the pathogenesis of COVID-19: Current and future perspectives

The coronavirus disease 2019 (COVID-19) pandemic underlines a persistent threat of respiratory tract infectious diseases and warrants preparedness for a rapid response. At present, COVID-19 has had a serious social impact and imposed a heavy global burden on public health. The exact pathogenesis of COVID-19 has not been fully elucidated. Since the outbreak of COVID-19, a renewed attention has been brought to Toll-like receptors (TLRs). Available data and new findings have demonstrated that the interaction of human TLRs and SARS-CoV-2 is a vital mediator of COVID-19 immunopathogenesis. TLRs such as TLR2, 4, 7 and 8 are potentially important in viral combat and activation of immunity in patients with COVID-19. Therapeutics targeting TLRs are currently considered promising options against the pandemic. A number of TLR-targeting immunotherapeutics are now being investigated in preclinical studies and different phases of clinical trials. In addition, innovative vaccines based on TLRs under development could be a promising approach for building a new generation of vaccines to solve the current challenges. In this review, we summarize recent progress in the role of TLRs in COVID-19, focusing the new candidate drugs targeting TLRs, the current technology and potential paths forward for employing TLR agonists as vaccine adjuvants.Copyright © 2023 The Scandinavian Foundation for Immunology.

Nirmatrelvir and Molnupiravir Coadministration Modifies Preclinical Viral Exposure

Background: Implementation of vaccination programmes has had a transformational impact on control of the SARS-CoV-2 pandemic, but the need for effective antiviral drugs remains. Molnupiravir (MPV) targets viral RNA polymerase inhibiting replication via lethal mutagenesis and nirmatrelvir (NTV) is a protease inhibitor boosted with ritonavir when given clinically. This study aimed to assess the virological efficacy of NTV and MPV individually and in combination against the SARS-CoV-2 BA.1 Omicron variant in a K18-hACE2 mouse model. Method(s): K18-hACE2 mice were inoculated intranasally with 103 PFU of SARSCoV-2 BA.1 Omicron (B.1.1.529). After 24 hours, mice were orally dosed q12H, as outlined in Figure 1. At 2, 3, and 4-days post infection mice were sacrificed, and lung samples harvested. Animals were weighed and monitored daily throughout. Subsequently, viral replication in the lung was quantified using qRT-PCR to measure total (N-gene) and sub-genomic (E-gene) viral RNA. Data were normalized to 18S for quantitation. Viral exposures expressed as Areas Under viral load Curves (AUCs) were calculated by the trapezoidal method using mean values at each timepoint. Separate studies in Syrian golden hamsters using individual drugs were also conducted, and total serum IgG was measured by ELISA at 4-days post infection. Result(s): Mice gained weight in all groups post-treatment, with no significant difference between groups. A reduction in lung viral exposure was evident in all treatment groups compared to the vehicle control dosed mice (Figure 1). Coadministration of NTV with MPV displayed a trend towards lower lung viral exposure compared to the vehicle control with ~40-and ~45-fold reduction in AUC for N-and SgE-gene assays, respectively. Dosed individually, NTV and MPV reduced viral exposure 5.7-and 7.7-fold for the N-gene assay, respectively. Differences in total serum IgG concentrations were evident between vehicle and NTV-(34-fold reduction, P=0.018), and MPV-(4.2-fold reduction, P=0.053) treated hamsters. Conclusion(s): These data show virological efficacy of NTV and MPV against the SARS-CoV-2 BA.1 Omicron variant. The combination of NTV and MPV demonstrated a lower viral RNA exposure in the lung than either drug alone, albeit not statistically significant. Initial data indicate potential immune alterations in NTV and MPV dosed hamsters. Studies to clarify the utility of NTV/ MPV combinations and further characterize the impact of antiviral therapy on IgG are warranted.

Utilization of prophylactic drugs for covid-19 infection among health care workers at government medical college Srinagar-a retrospective observational study

Introduction: Corona virus disease (COVID-19) was declared as a Pandemic by WHO on March 11, 2020. Since health care workers play an important role in providing care to infected patients, they are exposed to unprecedented levels of risk. At the initial phase of this pandemic, no definitive treatment was available, the only way to combat this disease was prevention. A number of prophylactic drugs were being studied during that time for use by health care workers. On 23rd March 2020, Government of India issued recommendation through National Task Force for Covid-19, for using Hydroxychloroquine as prophylaxis for SARS COV-2. Preclinical studies of Azithromycin have shown immunomodulation and in vitro activity against SARS-COV-2, that has led to its widespread usage during COVID-19. Ivermectin, an antiparasitic drug was reported to have an in vitro activity against SARS-COV-2. This orally administered drug was included in India's revised National COVID-19 treatment protocol for people with mild infection. Vitamin C, a water soluble vitamin has been considered for potential beneficial effects in COVID-19 disease. Many animal studies have indicated that a daily intake of vitamin C may prevent infections. Aim(s): To evaluate the pattern of drugs (HCQ, AZITHROMYCIN, IVERMECTIN,and VITAMIN C) used for COVID-19 prophylaxis among health care workers at GMC, Srinagar. MATERIALS AND METHODS: This study is being conducted by using a survey questionnaire. A survey questionnaire in English has been developed after literature review. The responses will be analyzed using descriptive statistics of frequency and percentage.

Accelerating antiviral discovery by combining machine learning with structural biology

COVID Moonshot is an international open science consortium aiming to discover oral antiviral against SARS-CoV-2, targeting the main protease. Launched in Feb 2020, Moonshot went from fragment hits to development candidates which are now under preclinical evaluation. In my talk, I will discuss Moonshot's journey, specifically how the combination of machine learning and structural biology has accelerated our design-make-test cycle. I will also discuss our vision for pandemic preparedness, and early results from AI-driven Structure Enabled Antiviral Platform (ASAP). ASAP is a NIH-funded antiviral drug discovery center which builds on COVID Moonshot's approach to target flaviviruses, enteroviruses, and coronaviruses. We are applying machine learning to generate potent chemical matter from crystallographic fragment hits, and leveraging high throughput library synthesis guided by models to rapidly expand on promising hits. Aiming to achieve pandemic preparedness, I will also discuss our approaches to preempting resistance, and how these strategic considerations impact drug-hunting.Copyright © 2023 The American Society for Biochemistry and Molecular Biology, Inc.

Overview: Scott M. Hammer Workshop for New Investigators and Trainees

Over the past four decades, remarkable progress has been made in understanding HIV epidemiology, pathogenesis, treatment, and prevention from the combined efforts of community members, clinicians, investigators, and funding agencies worldwide. Yet more work and new approaches are needed to achieve the ambitious goal of ending the epidemic and ensuring optimal quality of life for those living with HIV. To encourage and stimulate the next generation of investigators, the CROI Program Committee organizes an annual Workshop for New Investigators and Trainees comprised of expert and comprehensible talks to cover current knowledge and controversies in basic, clinical, and public health investigation into HIV and related infections, and to highlight relevant work to be presented over the ensuing days at CROI. This year, the program will begin with a presentation by Dr Stuart Neil on novel aspects of the molecular virology of HIV-1 and SARS-CoV-2. Following this, Dr Guido Silvestri will cover the immune responses against HIV and SARS-CoV-2. Ms Dawn Averitt, an HIV treatment policy advocate and activist will provide a community perspective on the power of community engagement in research. In the following presentation, Dr Monica Gandhi will review novel therapeutic strategies for HIV. Dr Raphael Landovitz will address advances in different strategies for preventing HIV transmission. Dr John Mellors will review advances in preclinical and clinical approaches for functional or sterilizing HIV-1 cure. The workshop will end with an intervention by Dr Rochelle Walensky, who will discuss career opportunities in research and public health. By completing the workshop, attendees will have achieved a head start toward maximizing the knowledge gained and research ideas arising from CROI 2022.

Epitope Masking during a Recall Response: Implications for Covid and Hiv Vaccines

Some of the first immunological experiments of the 20th century-on antibody feedback inhibition-demonstrated that past exposure does not merely expedite the humoral response to secondary challenges but, rather, alters the nature of that response. Despite this history, the mechanisms underpinning these classic observations and their relationship to modern vaccine design remain obscure. Circulating antibodies produced during a primary challenge can have enhancing or, conversely, inhibitory effects on later humoral responses. Using preclinical vaccine models, we dissected this apparent contradiction to find that the interaction between serum antibodies and the entry of cognate naive B cell lineages to germinal centers was determined by the interplay of breadth, affinity, and titer. Epitope-focused vaccine designs for HIV-1 elicit circulating antibodies capable of entirely obstructing their cognate naive B cells, with important implications for the ongoing design of boostphase immunogens. Conversely, SARS-CoV receptor binding domain (RBD) immunization elicits a polyclonal serum response that enhances the proportion of high-affinity cognate B cells in germinal centers, providing a partial explanation for the effectiveness of current boost protocols but also presaging diminishing returns unless new epitopes are introduced. The resolution of these surface-level contradictions points toward a unified interactive model and emphasizes that, to move vaccinology forward, the basic biology of the humoral immune system cannot remain a black box.

Using ex vivo bioengineered lungs to model pathologies and screening therapeutics

Introduction: More than 8 million lives are claimed annually by various respiratory diseases including lung cancer. While therapeutics is the first line of defence, treatment failure always remains challenging and research studies face a lag of transition from preclinical to clinical phase. This is partly due to the inadequate representation of the preclinical models in clinical trials. In this proof-of-concept study, we sought to use an ex-vivo model to identify lung pathologies and therapeutically screen them in a rodent model. Method(s): Briefly, the heart-lung tissues were extracted and decellularized using a detergent-based decellularization technique. Subsequently, lungs were seeded and cultured (6-10 days) with human cell lines: BEAS-2B, A549, and Calu3, demonstrating healthy lung, cancerous state, and congenital pathologies (cystic fibrosis), respectively. By altering the cultural conditions and exploiting the unique characteristics of these cell lines, we were able to model a variety of novel pathological models in ex vivo, such as advanced-stage solid tumours and the primary phase of infection via SARS-COV2. We also validated the above-mentioned observations by histology and immunofluorescence staining. Another novel part of our study includes a qualitative screening of efficacy and impact of important Therapeutics (anti-neoplastic)- Cisplatin and Wogonin, in our cancer models. Result(s): Using A549 and BEAS-2B cells, we were able to model different stages of Non-small cell lung cancer, qualitatively validated the resemblance to clinical samples and monitor the impact of different therapeutics on these models. The qualitative assessment also demonstrated different levels of cell death depending on the efficacy of the drugs. Contribution to research : Collectively this study demonstrates the remarkable versatility and strength of the ex vivo model in representing important lung pathologies and screening therapeutics in the preclinical phase.

Is Treating Post-Menopausal Women With Symptomatic Hand Osteoarthritis With Hormone Replacement Therapy Feasible?: Results From The Hope-E Randomised, Placebo-Controlled Feasibility Study

Purpose: Hand osteoarthritis (OA) is more common in women. Hand OA incidence increases further in females around the age of 50, the typical age of menopause. Estrogen-deficient states are associated with increased musculoskeletal pain and inflammation and with increased rates of symptomatic OA. Estrogen replacement and selective estrogen receptor modulators (SERMs) can improve pain and structure in some pre-clinical models of OA associated with estrogen loss, and in exploratory analysis from hormone replacement therapy (HRT) trials. However, no randomised clinical trials (RCTs) of HRT had been performed in symptomatic OA populations, specifically hand OA. By carrying out a RCT feasibility study of a form of HRT (conjugated estrogens (CE)-bazedoxifene) in post-menopausal women with painful hand OA, we set out to determine the feasibility and acceptability of this. We also aimed to generate proof-of concept data on likely outcomes, calculate a sample size and refine methods for a full trial. Method(s): We recruited females aged 40-65 years and 1-10 years after final menstrual period with definite hand OA and >=2 painful hand joints across three primary/secondary care sites and from the community. Medical exclusions included those typical for clinical HRT use. Design was parallel group, double-blind 1:1 randomisation of CE-bazedoxifene or placebo, taken orally once daily for 24 weeks, then tapering for 4 weeks before study end at Week 28. Primary feasibility outcomes were rates of eligible participant identification, recruitment, randomisation, retention, compliance, and likelihood of unblinding. Adverse events (AEs) were collected. Secondary clinical outcomes included the anticipated primary outcome in a full trial of mean hand pain over 14 days prior to each visit, scored on a 0-10 numerical rating scale (NRS) where 10 is worst pain possible, as well as hand function, appearance and menopause symptoms. Progression criteria to a full RCT were: (i) recruitment >=30 participants across all sites in 18 months (or proportionate to time open);(ii) a drop-out rate of <=30% of randomised individuals;and (iii) acceptability to the majority of participants, including acceptable AE rates. All clinical outcomes were analysed on an intention-to-treat basis. Though not powered to detect a treatment difference, change and treatment effects (the difference in the outcome between the two groups) were indicated with 95% CIs, with all models adjusted for clinical subtype of painful hand joint, study site, and baseline values. The sample size for a full trial was estimated using the standard deviation (SD) of week 24 mean hand pain. Result(s): Due to the COVID-19 pandemic, the recruitment window was reduced to 12-15 months. From May 2019 to December 2020, 434 enquiries/referrals were received. Of 96 telephone pre-screens, 35 individuals were potentially eligible and of these, 33 gave consent to participate. Of the remaining, 250/401 (62%) were ineligible, whilst 55/401 (14%) chose not to proceed, with the most common reason being not wanting to take HRT. 28/35 (80% (95%CI 63%,92%)) eligible participants were randomised to study medication. All 28 participants completed all follow-ups with high compliance (100% active, 13/14[93%] placebo) and outcome measure completeness (100%, mean hand pain). All three AE-related treatment withdrawals were on placebo when unblinded. No serious AEs occurred. Participants/investigators were well blinded (participant blinding index 0.50[95%CI 0.25 to 0.75]). All three prespecified progression criteria were therefore met for a full trial. The treatment effect difference over 24 weeks in mean hand pain between active and placebo was -0.71 (95% CI -2.20 to 0.78) (Fig 1A). During tapering/stopping medication, mean hand pain increased by 1.31 points in the active arm compared with 0.17 in the placebo arm, indicating a possible effect of cessation of medication (Fig 1A). Furthermore, 6/13 (46%) participants in the active group reported worsening pain at week 28 compared with week 24, but only 2/12 (17%) were worse on withdrawing placebo (Fig 1B). The sample size for a full trial was estimated as 296 (based on MCID 0.8 on NRS, SD 2.0, 90% power, 10% drop-out, alpha 5%). Conclusion(s): This first study of a RCT of HRT for painful hand OA met its progression criteria, indicating that a full trial of an HRT in this population is feasible and acceptable. Although not powered to detect an effect, there was a trend towards improvement in hand pain on treatment and worsening of hand pain on tapering in the active arm only. This adds to proof-of-concept data in this area, justifying more work.ISRCTN12196200. Funded by Research for Patient Benefit programme, National Institute for Health Research (UK) PB-PG-0416-20023 [Formula presented]Copyright © 2023

Morphological, Cellular, and Molecular Basis of Brain Infection in COVID-19 Patients and Its Association to Psychiatric Symptoms

Background: Although increasing evidence confirms neuropsychiatric manifestations associated mainly with severe COVID-19 infection, long-term neuropsychiatric dysfunction (recently characterized as part of "long COVID-19" syndrome) has been frequently observed after mild infection. Method(s): We performed a broad translational investigation, employing brain imaging and cognitive tests in 81 living COVID-19 patients (mildly infected individuals) as well as flow cytometry, respirometry, microscopy, proteomics, and metabolomics in postmortem brain samples, and in preclinical in vitro and ex vivo models. Result(s): We observed orbitofrontal cortical atrophy, neurocognitive impairment, excessive fatigue and anxiety symptoms in living individuals. Postmortem brain tissue from 26 individuals who died of COVID-19 revealed histopathological signs of brain damage. Five individuals out of the 26 exhibited foci of SARS- CoV-2 infection and replication, particularly in astrocytes. Supporting the hypothesis of astrocyte infection, neural stem cell-derived human astrocytes in vitro are susceptible to SARS-CoV-2 infection through a non-canonical mechanism that involves spike-NRP1 interaction. SARS-CoV-2-infected astrocytes manifested changes in energy metabolism and in key proteins and metabolites used to fuel neurons, as well as in the biogenesis of neurotransmitters. Moreover, human astrocyte infection elicits a secretory phenotype that significantly reduces neuronal viability. Conclusion(s): Our data support the model in which COVID-19 alter cortical thickness, promoting psychiatric symptoms. In addition, SARS-CoV-2 is able to reach the brain, infects astrocytes, and consequently, leads to neuronal death or dysfunction. These deregulated processes could contribute to the structural and functional alterations seen in the brains of COVID-19 patients. Funding Source: Sao Paulo Research Foundation (FAPESP) Keywords: COVID-19, Anxiety, Astrocytes, Multi-omics, Brain Magnetic Resonance Imaging (MRI)Copyright © 2023

Evaluation of the immunogenicity of an inactivated vaccine prototype against SARS-COV- 2 virus in vitro

Background: Belarus started developing a vaccine against SARS-CoV- 2 in 2021. The aim of the first stage of investigation was to evaluate the immunogenicity of the vaccine prototypes (VP) in vitro. Method(s): SARS-CoV- 2 strains (n = 7) were isolated using Vero E6 cells, inactivated with beta-propiolactone and purified. Antigens (Ag) were adsorbed on adjuvants: Al(OH)3 (Ag+AH group) or Al3PO4 (Ag+AP group). The single dose of VP (500 ul) was composed of 10 mug of Ag adsorbed on adjuvants (200 mug of Al3+). Blood samples from SARS-CoV- 2 recovered donors (n = 18) and healthy controls having no history of COVID-19 infection (n = 5) were used. Whole blood and Tag-it Violet labeled PBMCs were cultivated with VP, pure Ag, adjuvants (0.25-1 mug of Ag, 20 mug of Al3+ for probe) or pool of peptides, covering sequence of SARS-CoV- 2 N, S, M-proteins (PP), for 6 h and 7 days respectively. INF-gamma production and proliferation of CD3+ T-cells were assayed by FACS. Result(s): Counts of CD3+IFN-gamma+ T cells were 3.14(2.72-5.13)/ 1x105 CD3+ T cells in negative control (NC), and 12.73(10.09-33.95)/ 1x105 CD3+ T cells in specific positive control (PP) (n = 18, p < 0.0001), proving presence of antigen-specific T cells (ASCs) in donor blood. Samples were considered positive for VP and Ag immunogenicity when numbers of CD3+IFN-gamma+ T cells were 1.5 times greater compared with NC. Both VP types (Ag+AP, Ag+AH) and pure SARS-CoV- 2 isolates stimulated the production of INF-gamma by ASCs, responses ranged from 1 to 4 isolates of 7 studied per donor. Immunogenicity of Ag+AP, Ag+AH was confirmed by proliferation assay. Proliferation level was 1.07(0.97-2.38)% in Ag group with no differences from NC (n = 7, p > 0.5). Proliferation was significantly greater in VP groups compared with Ag: 2.47(1.65-2.68)% in Ag+AH, 4.03(2.56-4.61)% in Ag+AP (p = 0.009 and 0.002, respectively), stimulation of T cell was stronger by Ag+AP compared with Ag+AH (p = 0.009, M-U test). Pure adjuvants did not induce T cells response. There was no T-cell stimulation by Ag and VP in samples obtained from COVID-19 negative donors. Conclusion(s): The VP against SARS-CoV- 2 infection composed of inactivated virus adsorbed on Al(OH)3 and Al3PO4 adjuvants has immunogenic properties proven in two different immunological assays. VP stimulated activation and proliferation of ASCs in vitro suggesting this VP can be used for further preclinical in vivo evaluation.

Dataset on safety and protective efficacy studies of COVID-19 vaccine candidates based on structurally modified plant virus in female hamsters.

This data article is related to the previous research, which addressed the development of a COVID-19 recombinant vaccine candidate. Here, we present the additional data in support of the safety and protective efficacy evaluation of two COVID-19 vaccine candidates based on the coronaviruses' S protein fragments and a structurally modified plant virus - spherical particles. The effectiveness of the experimental vaccines was studied against the SARS-CoV-2 virus in an in vivo infection model in female Syrian hamsters. The body weight of vaccinated laboratory animals was monitored. The histological assessment data of the infected with the SARS-CoV-2 virus hamsters' lungs are provided.

Daylight photodynamic therapy at home

Daylight photodynamic therapy (DPDT) is an established treatment for field-change actinic keratoses (AK) with high rates of satisfaction and success. In recent times there has been a push within the healthcare industry to reduce avoidable clinic time and complement it with community-based healthcare, including self-administration therapies. The importance of 'decentralized' healthcare and at-home therapies has been emphasized by the recent COVID-19 pandemic - access to treatments is restricted and many patients are not receiving the appropriate care in an attempt to minimize hospital-based treatments. In this project, we deconstructed DPDT and by utilizing principles of design and the concept of realistic medicine, transformed it into a user-friendly, environmentally conscious and engaging at-home therapeutic option. Information on protocols and best practice was obtained from clinical colleagues and a map of the patient pathway was outlined. The treatment was broken down and re-formed into simple steps, taking care with the number of instructions to prevent confusion. The physical form of the at-home kit was designed to facilitate the required materials for DPDT, while being simple and methodical to follow. Steps were separated into individual numbered sections, with only the materials needed at each step visible. Simple graphics are displayed alongside relevant instructions, with colouring to highlight importance. The at-home kit was iteratively improved with input from end users. As part of this initiative the DPDT athome kit is designed and prototyped in order to be posted directly to the user. In trialling this kit preclinically, the theoretical patient journey could be visualized, starting with the unboxing of the kit, then followed by the guides and directed procedure. Through feedback, iterations to the design have subsequently been made that efficiently translate the clinical procedure into a successful at-home design. One of the key principles of realistic medicine to consider is the reduction of waste. In this kit we have, where possible, used recycled and recyclable materials, and are in the process of incorporating medically approved biodegradable gloves, which will instantly reduce a high fraction of the nonrecyclable excess. Implementation of the kit in routine clinical practice will provide important feedback allowing further iterations to the design of the kit. Involving patients directly with the development work and continuously responding to the patient experience will significantly improve the final design of the at-home kit. Helping to implement an option to take this important treatment away from a hospitalized environment represents a paradigm shift in the possible delivery of DPDT and can be useful to optimize treatment delivery on a per-patient basis.

Discovery of Potential Inhibitors of SARS-CoV-2 (COVID-19) Main Protease (Mpro) from Nigella Sativa (Black Seed) by Molecular Docking Study

Purpose: A new human coronavirus (SARS-CoV-2), triggering pneumonia, is termed as Coronavirus Disease-19 (COVID-19). There is an alarming situation now as this new virus is spreading around the world. At present, there are no specific treatments for COVID-19. Nigella sativa is known as Prophetic Medicine as its use has been mentioned in Prophetic Hadith, as a natural remedy for all the diseases except death. Seeds and oils of N. sativa have a long history of folklore usage in various systems of medicine such as Unani and Tibb, Ayurveda and Siddha in the treatment of different diseases and ailments. The aim of this research is to provide a potential inhibitor of SARS-CoV-2 Mpro. Method(s): The Molecular docking tool was used to optimize the binding affinities of chemical constituents of N. sativa with SARS-CoV-2 Mpro. Result(s): Many constituents from N. Sativa have shown better binding affinity than reported drugs with SARS-CoV-2 Mpro i.e., the alpha-hederin, Stigmasterol glucoside, Nigellidine-4-O-sulfite, Nigellidine, Sterol-3-beta-D-glucoside, Dithymoquinone, beta-sitosterol have binding affinities (kcal/mol) of-9,-8.1,-8,-7.7,-7.7,-7.4,-7.4, and-6.9 and number of hydrogen bonds formed are 06, 04, 03, 03, 03, 00, and 01, respectively. Conclusion(s): There is rationale and pre-clinical evidence of the effectiveness of N. Sativa that it may be helpful for the treatment of COVID-19 and can serve as a potential natural candidate. However, more studies should be conducted to collect high-quality data and scientific evidence of N. Sativa to use it against COVID-19 clinically.Copyright © 2021 Bentham Science Publishers.

Neurovirulence of SARS CoV2: From Clinical Data to Preclinical Neu-ropsychological Exploration

Introduction: COVID-19 pandemic represents a major health issue caused by SARS CoV2, a human coronavirus. Since the outbreak of this pandemic, the literature on SARS CoV-2 has grown differentially, with increased awareness of extra-respiratory symptoms, including neurological symptoms. Method(s): Review based on studies published from December 2019 to June 2020. Results and Discussion: This review discusses the neurological aspect of SARS CoV2, including the suggested mechanism involved. Neurological disorders are cited in addition to emerging experimental models with viral involvement. Conclusion(s): There is a need for further investigation to clarify how it can lead to the onset of acute and chronic neurological disorders, mentioning the importance of experimental studies in neuropsy-chopharmacology.Copyright © 2022 Bentham Science Publishers.

The Outbreak of Coronavirus Disease 2019 (COVID-19) and its Manifestation

The outbreak of coronavirus disease 2019 (COVID-19) was first reported in Wuhan, Chi-na, and soon the infection turned into a pandemic. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the spread of COVID-19 infection. The World Health Organization (WHO) confirmed 4.20% of total deaths globally (March 21, 2020). Within four months (July 21, 2020), the rate of confirmed total deaths was recorded up to 4.17% globally. In India, 909 confirmed cases and 19 deaths were reported by Health and Family Welfare, Government of India, on March 28, 2020. Over a period of 123 days in India, 1638870 confirmed cases and 35684 deaths were reported. COVID-19 can potentially spread from person to person through direct contact or respiratory droplets from coughing and sneezing. The most common symptoms are fever, dry cough, difficulty in breathing, and fatigue. A pregnant mother with COVID-19 has fewer chances of trans-ferring this infection to her newborn baby. Children are less affected than adults from this virus. A specific antiviral drug or vaccine has not been developed to cure the disease to date. Drugs including chloroquine, hydroxychloroquine, lopinavir, ritonavir, nafamostat, nitazoxanide, and remde-sivir, have been observed to be effective for treating COVID-19. Many vaccine candidates are under investigation in pre-clinical and clinical studies. In this review, we highlight the epidemiology, signs and symptoms, pathogenesis, and mode of transmission of the infection caused by COVID-19, and its effects on a pregnant mother and newborn, and children. We also highlight the preventive measures and drugs that are effective for treating COVID-19.Copyright © 2021 Bentham Science Publishers.

Current Trends and Future Strategies for the Global Impact of COVID-19 Pandemic

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) might have originated from the recombination of a Pangolin-CoV-like virus with a Bat-CoV-RaTG13-like virus and then transmitted to a human at Wuhan city of China. On February 11, 2020, the WHO announced a name for the new coronavirus disease as COVID-19. Finally, the WHO declared the novel coron-avirus outbreak a global pandemic on March 11, 2020. Within a few months, SARS-CoV-2 had spread across the world to 220 countries, areas or territories. The main objective of this work is to review the existing knowledge about COVID-19, its updated status, available treatment procedures and future challenges. The available literature based on the COVID-19 was thoroughly reviewed and concise, evidence-based information was explored for the public interest. Various authentic databases like PubMed, Scopus, and Google Scholar together with the official sites of some Govt. Organizations were carefully searched for all relevant information about the current status of COVID-19, including the published research on coronavirus. More than 68 million people are already infected, including around 20% severely ill, with almost 1.5 million casualties due to this virus which is expected to infect approximately 70% population worldwide. Currently, maximum confirmed cases and death are reported in the USA. The epicentre of the pandemic was initially shifted from China to Europe, then to the USA, Brazil and now India. In between, the understanding of pathogenesis and mode of transmission has been developed;repurposing drugs are being validated and the development of a new vaccine is underway. The study concludes that there is no established treatment available for COVID-19, although 26 clinical and 139 preclinical trials are un-derway to develop vaccines globally. Although three vaccines are at the advanced stage of develop-ment, their efficacy and adverse effects are yet to be validated and recorded. Recently, the Pfizer vaccine has been started for vaccination in emergency cases in England and Bahrain, and the United States of America will start it soon. Meanwhile, prevention, rigorous global containment and quarantine efforts are practiced worldwide to control its spread.Copyright © 2021 Bentham Science Publishers.