28-day repeated dose toxicological evaluation of Coronil in Sprague Dawley rats: Behavioral, hematological, biochemical and histopathological assessments under GLP compliance.

Ayurvedic medicines are widely employed globally for prophylaxis and treatment of a variety of diseases. Coronil is a tri-herbal medicine, constituted with the traditional herbs, Tinospora cordifolia, Withania somnifera and Ocimum sanctum, with known immunomodulatory activities. Based on its proven in-vitro activity and in-vivo efficacy, Coronil has been approved as a 'Supporting Measure for COVID-19' by the Ministry of AYUSH, Government of India. The current study was aimed to assess the non-clinical safety of Coronil in a 28-day repeated dose toxicity study along with a 14-day recovery period in Sprague Dawley rats. This toxicity study was conducted in accordance with OECD test guideline 407, under GLP-compliance. Specific-Pathogen-Free animals of either sex, housed in Individually-Ventilated-Cages were particularly used in the study. The tested Coronil dose levels were 0, 100, 300 and 1000 mg/kg/day, orally administered to 5 males and 5 female rats per test group. In the current study, no mortality was observed in any group and in addition, Coronil did not elicit any finding of toxicological relevance with respect to clinical signs, ocular effects, hematology, urinalysis and clinical chemistry parameters, as well as macro- or microscopical changes in any organs, when compared to the control group. Accordingly, the No-Observed-Adverse-Effect-Level (NOAEL) of Coronil was ascertained to be 1000 mg/kg/day, subsequent to its 28-day oral administration to male and female rats. The acceptable safety profile of Coronil paves the way further toxicity assessments in rodents for a longer duration as well as in higher animals, and towards its clinical investigation.

Phyto-Compounds from a Rather Poisonous Plant, Strychnos nuxvomica, Show High Potency Against SARS-CoV-2 RNA-Dependent RNA Polymerase.

BACKGROUND: The establishment of strategy to inhibit the virus replication is an attractive means in combating SARS-CoV-2 infection. OBJECTIVE: We studied phyto-compounds from Strychnos nux-vomica (a poisonous plant) against SARS-CoV-2 RNA-dependent RNA polymerase by computational methods. METHODS: Molecular docking, molecular dynamics (MD) simulation and energetics calculations were employed to elucidate the role of the phyto-compounds. RESULTS: Ergotamine with a binding free energy of -14.39 kcal/mol showed a promising capability in terms of binding affinity and the interaction to conserved motifs, especially the SDD signature sequence. The calculated dissociation constants for ATP, ergotamine, isosungucine and sungucine were 12 µM, 0.072 nM, 0.011 nM and 0.152 nM, respectively. The exhibited kd by these phyto-compounds reflected tens of thousands fold potency as compared to ATP. The binding free energies of sungucine and isosungucine were much lower (-13.93 and -15.55 kcal/mol, respectively) compared to that of ATP (-6.98 kcal/mol). CONCLUSION: Sharing the same binding location as that of ATP and having high binding affinities, Ergotamine, Isosungucine, Sungucine and Strychnine N-oxide could be effective in controlling the SARS-CoV-2 virus replication by blocking the ATP and inhibiting the enzyme function.

Psychological Impacts of COVID-19 in Dental Patients are Moderated and Mediated by Hospital-Infection-Control-Policy and Satisfaction-with-Life: A Prospective Observational Dental-COVID Study.

Introduction: COVID-19 pandemic has imposed nation-wide lock-downs which severely impacted day-to-day lifestyle and caused anxiety, stress and fear among patients taking medical care including dental treatments. These psychological behaviors have also been observed during the strategic relaxation of social restrictions (Unlock). However, potential effect of these psychological behavior in endodontic cases have not been probed. Here, it is of great interest to explore the magnitude and buffering effect of two important psychological resources: satisfaction-with-life, and confidence in hospital-infection-control-policy in relation with COVID-19 risk perception on psychological impact due to fear for COVID-19. Methods: Patients visiting Dental Clinic & Research Center for endodontic procedures were randomly asked to fill survey questionnaire, and were later enrolled as per the study criteria. The study carried out in two phases: from 15 Sept 2020 to 15 Dec 2020 (restrained confinement; Unlock 1.0); and from 16 Dec 2020 to 12 Feb 2021 (mild confinement; Unlock 2.0) with total sample size of 136. We used data collection tools such as fear-of-COVID-19 scale (FCV-19), perceived-stress-scale (PSS), modified-dental-anxiety-scale (MDAS), satisfaction-with-life scale (SWLS), COVID-19 risk perception, and confidence in hospital-infection-control-policy for COVID-19. Results: A double moderation and dual moderated mediation structured model were used to establish the correlation of various parameters using SPSS (version 25.0) software suite. Confidence in hospital-infection-control-policy and SWLS were negatively correlated with FCV-19, MDAS, and PSS. Risk perception of COVID-19 was found to positively associated with FCV-19, MDAS, and PSS. Discussion: Patient's confidence in hospital-infection-control-policy for COVID-19 and SWLS acted as independent moderator for FCV-19 and mental distress. FCV-19 and risk perception were found to be lower during mild confinement (Unlock 2.0), and were also the positive predictors of PSS; and negative predictors of SWLS. The higher SWLS correlated very well with lower COVID-19 risk perception, concerning PSS and MDAS.

Germi-X herbal-based spray disinfects smartphone surfaces: implication on fomite-mediated infection spread.

Inanimate objects/surfaces become fomites upon contacting infectious agents such as disease-causing bacteria, fungi and viruses. Smartphones are one of the most prominent among these fomites. COVID-19 pandemic has raised the awareness on mobile sanitization, as an active measure to curb fomite-mediated viral transmission. Available mobile sanitizers and ultraviolet (UV) ray mediated mobile sanitization have their own sets of pros and cons, often being less user-friendly. This study explored the germicidal efficacy of an herbal-based sanitizer, Germi-X spray, on hands and mobiles, through microbiological techniques of micro-broth dilution and Kirby-Bauer disc diffusion assay, thumb print assay and swab test. Notably, Germi-X spray was found to be 6-67% more effective against surface pathogens, like, Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas fluorescens and Pseudomonas aeruginosa, as compared to a very popular product in the Indian market, which was taken as a control for this study. The observed anti-bacterial activity of the spray from disc-diffusion assay suggests its greater surface retentivity as compared to the control. Germicidal potency of Germi-X spray, when used to sanitize hands, was found to be greater than 80%. There was ~ 17-fold reduction in microbial counts after sanitizing smartphones with Germi-X spray. The novelty of this study lies in providing experimental evidence for this herbal-based surface sanitizer in efficiently disinfecting one of the super contaminated fomite, the smartphones. In conclusion, having an herbal base with a high germicidal efficacy against surface pathogens, together with longer surface retention, Germi-X spray appears to be an eco-friendly and cost-effective sanitizer for the surfaces of electronic gadgets like smartphones.

Phytometabolite profiling of Coronil, a herbal medicine for COVID-19, its identification by mass-spectroscopy and quality validation on liquid chromatographic platforms.

Coronil is a tri-herbal medicine consisting of immunomodulatory herbs, Withania somnifera, Tinospora cordifolia, and Ocimum sanctum. The formulation has been developed specifically as the supporting measure for COVID-19. Current investigation is aimed to identify the phytoconstituents in Coronil utilizing ultra-performance liquid chromatography-mass spectrometry coupled with quadrapole time of flight and to establish its quality standardization using high-performance liquid chromatography and high performance thin layer chromatography. Out of 52 identified compounds, cordifolioside A, magnoflorine, rosmarinic acid, palmatine, withanoside IV, withanoside V, withanone, betulinic acid, and ursolic acid were quantified in 15 different batches of Coronil on validated high-performance liquid chromatography method. Similarly, withanoside IV, withaferin A, magnoflorine, palmatine, rosmarinic acid, and ursolic acid were analyzed on high performance thin layer chromatography. Methods were validated as per the International Council for Harmonization guidelines. These methods were specific, reproducible, accurate, precise, linear (r2 > 0.99), and percent recoveries were within the prescribed limits. The content uniformity of Coronil was ascertained using Fourier transform infrared spectroscopy. Results indicated that, validated methods were fit for their intended use and the analytical quality of Coronil was consistent across the batches. Taken together, these developed methods could drive the analytical quality control of herbal medicines such as Coronil, and other formulations containing similar chemical profiles.

Yoga instructors' reported benefits and disadvantages associated with functioning online: A convenience sampling survey.

BACKGROUND: Among numerous changes in response to the COVID-19 pandemic, most yoga classes have repositioned online. However benefits, difficulties and satisfaction of teaching yoga online remain to be studied. With this background the present survey aimed to determine: (i) benefits, disadvantages and satisfaction of teaching yoga online and (ii) their association with characteristics related to (a) socio-demographic, (b) online yoga teaching experience and (c) yoga practice. METHODS: Three hundred and five yoga instructors were invited to take part in the online survey. Of these, 181 (m:f = 98:83) responded to the survey satisfactorily and were included. RESULTS: The three most common benefits of teaching yoga online were: (i) a sense of safety from risk of COVID-19 (93.92%), (ii) cost saving (82.87%) and (iii) wider access to trainees within India (77.90%). The three most common disadvantages were: (i) technical difficulties (74.03%), (ii) missing in-person contact (63.90%) and (iii) concern that online instructions can lead to injury (59.16%). Around 66.30% respondents were satisfied with the monitoring of trainees during online yoga classes while 70.16% respondents were satisfied with the level of attention they could pay to the topic they were teaching during online yoga class. The benefits and disadvantages of teaching yoga online varied with the characteristics of yoga instructors (p < 0.05, χ2 test). CONCLUSIONS: The benefits and disadvantages of teaching yoga online are of relevance during and beyond the pandemic. Characteristics related to (i) socio-demographics, (ii) online yoga teaching and (iii) yoga practice influence reported benefits and disadvantages of teaching yoga online.

Nanotechnology Interventions in the Management of COVID-19: Prevention, Diagnosis and Virus-Like Particle Vaccines.

SARS-CoV-2 claimed numerous lives and put nations on high alert. The lack of antiviral medications and the small number of approved vaccines, as well as the recurrence of adverse effects, necessitates the development of novel treatment ways to combat COVID-19. In this context, using databases such as PubMed, Google Scholar, and Science Direct, we gathered information about nanotechnology's involvement in the prevention, diagnosis and virus-like particle vaccine development. This review revealed that various nanomaterials like gold, polymeric, graphene and poly amino ester with carboxyl group coated magnetic nanoparticles have been explored for the fast detection of SARS-CoV-2. Personal protective equipment fabricated with nanoparticles, such as gloves, masks, clothes, surfactants, and Ag, TiO2 based disinfectants played an essential role in halting COVID-19 transmission. Nanoparticles are used not only in vaccine delivery, such as lipid nanoparticles mediated transport of mRNA-based Pfizer and Moderna vaccines, but also in the development of vaccine as the virus-like particles elicit an immune response. There are now 18 virus-like particle vaccines in pre-clinical development, with one of them, developed by Novavax, reported being in phase 3 trials. Due to the probability of upcoming COVID-19 waves, and the rise of new diseases, the future relevance of virus-like particles is imperative. Furthermore, psychosocial variables linked to vaccine reluctance constitute a critical problem that must be addressed immediately to avert pandemic.

Application of Humanized Zebrafish Model in the Suppression of SARS-CoV-2 Spike Protein Induced Pathology by Tri-Herbal Medicine Coronil via Cytokine Modulation.

Zebrafish has been a reliable model system for studying human viral pathologies. SARS-CoV-2 viral infection has become a global chaos, affecting millions of people. There is an urgent need to contain the pandemic and develop reliable therapies. We report the use of a humanized zebrafish model, xeno-transplanted with human lung epithelial cells, A549, for studying the protective effects of a tri-herbal medicine Coronil. At human relevant doses of 12 and 58 µg/kg, Coronil inhibited SARS-CoV-2 spike protein, induced humanized zebrafish mortality, and rescued from behavioral fever. Morphological and cellular abnormalities along with granulocyte and macrophage accumulation in the swim bladder were restored to normal. Skin hemorrhage, renal cell degeneration, and necrosis were also significantly attenuated by Coronil treatment. Ultra-high-performance liquid chromatography (UHPLC) analysis identified ursolic acid, betulinic acid, withanone, withaferine A, withanoside IV-V, cordifolioside A, magnoflorine, rosmarinic acid, and palmatine as phyto-metabolites present in Coronil. In A549 cells, Coronil attenuated the IL-1ß induced IL-6 and TNF-α cytokine secretions, and decreased TNF-α induced NF-κB/AP-1 transcriptional activity. Taken together, we show the disease modifying immunomodulatory properties of Coronil, at human equivalent doses, in rescuing the pathological features induced by the SARS-CoV-2 spike protein, suggesting its potential use in SARS-CoV-2 infectivity.

Development and validation of a rapid high-performance thin-layer chromatographic method for quantification of gallic acid, cinnamic acid, piperine, eugenol, and glycyrrhizin in Divya-Swasari-Vati, an ayurvedic medicine for respiratory ailments.

Divya-Swasari-Vati is a calcium containing polyherbal ayurvedic medicine prescribed for the lung-related ailments observed in the current pandemic of Severe Acute Respiratory Syndrome Coronavirus 2 infections. The formulation is a unique quintessential blend of nine herbs cited in Ayurvedic texts for chronic cough and lung infection. Analytical standardization of herbal medicines is the pressing need of the hour to ascertain the quality compliance. This persuaded us to develop a simple, rapid, and selective high-performance thin-layer chromatographic method for Divya-Swasari-Vati quality standardization. The developed method was validated for the quantification of marker components, gallic acid, cinnamic acid, piperine, eugenol and glycyrrhizin, against reference standards in five different batches of Divya-Swasari-Vati. The analytes were identified by visualization at 254 nm, and by matching their retention factor with authentic standards. The developed method was validated as per the guidelines recommended by the International Council for Harmonization for parameters like, linearity, limit of detection, limit of quantification, accuracy, and precision. Therefore, the developed novel high-performance thin-layer chromatographic process could be employed for rapid standardization of Divya-Swasari-Vati and other related herbal formulation, which would aid in quality manufacturing and product development.

Comparative retrospective open-label study of ayurvedic medicines and their combination with allopathic drugs on asymptomatic and mildly-symptomatic COVID-19 patients.

INTRODUCTION: Treatment for COVID-19 was ambiguous in the beginning of the pandemic. At that time, the conventional medical system was grappling to cope with the rapidly spreading pandemic. The potential of Ayurveda, one of the branches of traditional Indian medicine (TIM), with a 5000 year old history, employing medicines derived from plants and other natural sources, against COVID-19 has been explored through a comparative retrospective open-label study. METHODS: Reported here are the remedial effects of Ayurvedic medicines alone or in combination with Allopathic treatment on 59 asymptomatic or mildly symptomatic COVID-19 patients, across multiple COVID-19 care centers in Ahmedabad, India. The patients were confirmed for COVID-19 infection through RT-qPCR of nasopharyngeal swabs. With informed consents from the patients, the sourced data was divided into 'Allopathic and Ayurvedic' [AlloAyur] (n = 41) and 'Ayurvedic only'[Ayur] (n = 18) groups, based on the type of treatment the patients decided to receive, that is Ayurvedic medicines with Allopathic treatment or Ayurvedic medicines alone, respectively. Ayurvedic medicines included oral doses and nasal drops; the dosage and regime were decided based on the recommendations from Ayurvedic texts. The Allopathic medicines included Azithromycin, Vitamin-C and anti-histamines. Acetaminophen was also administered when necessary, by the attending physician. The patients were observed for symptomatic improvement. RESULTS: Primary outcome of this study was the symptomatic relief from COVID-19. Data collected over a period of two months, showed that more patients exhibited symptomatic relief in Ayur goup (83.33 %) than in the AlloAyur group (48.78 %) within the first 13 days of treatment. No visible adverse effects were observed. This indicated faster and safe symptomatic resolution among those treated with Ayurvedic medicines alone. CONCLUSION: Patients receiving only Ayurvedic medicines on average were symptomatically relieved faster than those receiving Allopathic and Ayurvedic medicines together.

Influence of Patient-Reported Treatment Satisfaction on Psychological Health and Quality of Life Among Patients Receiving Divya-Swasari-Coronil-Kit Against COVID-19: Findings from a Cross-Sectional "SATISFACTION COVID" Survey.

INTRODUCTION: The correlation among treatment satisfaction with demographic characteristics, health symptoms or psychological health, and quality of life with the prophylactic regime against COVID-19 is rather unexplored. This real-world exploratory study was conducted to determine patient-perspectives regarding their treatment satisfaction receiving Divya-Swasari-Coronil-Kit with correlative impacts on psychological health (PH) and Quality of life (QoL) based on four hypotheses each relating to PH, QoL, Demographic characteristics, and Treatment satisfaction. METHODS: This cross-sectional, web-based survey collected data on demographic characteristics and psychological health with DASS-21; QoL with 5-level 5-dimension EuroQol instrument; and treatment satisfaction using Treatment Satisfaction Questionnaire for Medication (TSQM) V9. Pearson correlation coefficient analysis was used to examine the relation between TSQM and PH and the demographic variables. Factor analysis was used for multi-collinearity tests, and multiple linear regression analysis was used to explore demographic variables and TSQM. RESULTS: Out of 421 initial screenings, 367 patient-participants were included in the analysis. The mean age of included participants was 33.61 ± 9.47 years. Marital status and socio-economic class positively correlated with TSQM. Physical symptoms in patients are positively correlated with depression, anxiety, and stress; and in contrast, negatively with QoL. Global satisfaction with Divya-Swasari-Coronil-Kit medication negatively correlated with depression, anxiety, stress, effectiveness, convenience; whereas global satisfaction correlated positively with QoL. CONCLUSION: Present study (SATISFACTION COVID) indicates that treatment satisfaction due to avaliablity and treatment of Divya-Swasari-Coronil-Kit has constructive and beneficial implications on psychological health, Quality of life and demographic factors. In addition, web-based patient-reported perspectives may well be a feasible way to provide better insights into treatment satisfaction, in relation to psychological health and Quality of life.

Giloy Ghanvati (Tinospora cordifolia (Willd.) Hook. f. and Thomson) Reversed SARS-CoV-2 Viral Spike-Protein Induced Disease Phenotype in the Xenotransplant Model of Humanized Zebrafish.

The current Severe Acute Respiratory Syndrome disease caused by Coronavirus-2 (SARS-CoV-2) has been a serious strain on the healthcare infrastructure mainly due to the lack of a reliable treatment option. Alternate therapies aimed at symptomatic relief are currently prescribed along with artificial ventilation to relieve distress. Traditional medicine in the form of Ayurveda has been used since ancient times as a holistic treatment option rather than targeted therapy. The practice of Ayurveda has several potent herbal alternatives for chronic cough, inflammation, and respiratory distress which are often seen in the SARS-CoV-2 infection. In this study we have used the aqueous extracts of Tinospora cordifolia (willd.) Hook. f. and Thomson in the form of Giloy Ghanvati, as a means of treatment to the SARS-CoV-2 spike-protein induced disease phenotype in a humanized zebrafish model. The introduction of spike-protein in the swim bladder transplanted with human lung epithelial cells (A549), caused an infiltration of pro-inflammatory immune cells such as granulocytes and macrophages into the swim bladder. There was also an increased systemic damage as exemplified by renal tissue damage and increased behavioral fever in the disease induction group. These features were reversed in the treatment group, fed with three different dosages of Giloy Ghanvati. The resultant changes in the disease phenotype were comparable to the group that were given the reference compound, Dexamethasone. These findings correlated well with various phyto-compounds detected in the Giloy Ghanvati and their reported roles in the viral disease phenotype amelioration.

Comprehensive and Rapid Quality Evaluation Method for the Ayurvedic Medicine Divya-Swasari-Vati Using Two Analytical Techniques: UPLC/QToF MS and HPLC-DAD.

Divya-Swasari-Vati (DSV) is a calcium-containing herbal medicine formulated for the symptomatic control of respiratory illnesses observed in the current COVID-19 pandemic. DSV is an Ayurvedic medicine used for the treatment of chronic cough and inflammation. The formulation has shown its pharmacological effects against SARS-CoV-2 induced inflammation in the humanized zebrafish model. The present inventive research aimed to establish comprehensive quality parameters of the DSV formulation using validated chromatographic analytical tools. Exhaustive identification of signature marker compounds present in the plant ingredients was carried out using ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/QToF MS). This was followed by simultaneous estimation of selected marker components using rapid and reliable high-performance liquid chromatography (HPLC) analysis. Eleven marker components, namely gallic acid, protocatechuic acid, methyl gallate, ellagic acid, coumarin, cinnamic acid, glycyrrhizin, eugenol, 6-gingerol, piperine and glabridin, were selected out of seventy-four identified makers for the quantitative analysis in DSV formulation. Validation of the HPLC method was evaluated by its linearity, precision, and accuracy tests as per the International Council of Harmonization (ICH) guidelines. Calibration curves for the eleven marker compounds showed good linear regression (r2 > 0.999). The relative standard deviation (RSD) value of intraday and interday precision tests were within the prescribed limits. The accuracy test results ranged from 92.75% to 100.13%. Thus, the present inclusive approach is first of its kind employing multi-chromatographic platforms for identification and quantification of the marker components in DSV, which could be applied for routine standardization of DSV and other related formulations.

Withanone from Withania somnifera Attenuates SARS-CoV-2 RBD and Host ACE2 Interactions to Rescue Spike Protein Induced Pathologies in Humanized Zebrafish Model.

PURPOSE: SARS-CoV-2 engages human ACE2 through its spike (S) protein receptor binding domain (RBD) to enter the host cell. Recent computational studies have reported that withanone and withaferin A, phytochemicals found in Withania somnifera, target viral main protease (MPro) and host transmembrane TMPRSS2, and glucose related protein 78 (GRP78), respectively, implicating their potential as viral entry inhibitors. Absence of specific treatment against SARS-CoV-2 infection has encouraged exploration of phytochemicals as potential antivirals. AIM: This study aimed at in silico exploration, along with in vitro and in vivo validation of antiviral efficacy of the phytochemical withanone. METHODS: Through molecular docking, molecular dynamic (MD) simulation and electrostatic energy calculation the plausible biochemical interactions between withanone and the ACE2-RBD complex were investigated. These in silico observations were biochemically validated by ELISA-based assays. Withanone-enriched extract from W. somnifera was tested for its ability to ameliorate clinically relevant pathological features, modelled in humanized zebrafish through SARS-CoV-2 recombinant spike (S) protein induction. RESULTS: Withanone bound efficiently at the interacting interface of the ACE2-RBD complex and destabilized it energetically. The electrostatic component of binding free energies of the complex was significantly decreased. The two intrachain salt bridge interactions (K31-E35) and the interchain long-range ion-pair (K31-E484), at the ACE2-RBD interface were completely abolished by withanone, in the 50 ns simulation. In vitro binding assay experimentally validated that withanone efficiently inhibited (IC50=0.33 ng/mL) the interaction between ACE2 and RBD, in a dose-dependent manner. A withanone-enriched extract, without any co-extracted withaferin A, was prepared from W. somnifera leaves. This enriched extract was found to be efficient in ameliorating human-like pathological responses induced in humanized zebrafish by SARS-CoV-2 recombinant spike (S) protein. CONCLUSION: In conclusion, this study provided experimental validation for computational insight into the potential of withanone as a potent inhibitor of SARS-CoV-2 coronavirus entry into the host cells.

Coronil, a Tri-Herbal Formulation, Attenuates Spike-Protein-Mediated SARS-CoV-2 Viral Entry into Human Alveolar Epithelial Cells and Pro-Inflammatory Cytokines Production by Inhibiting Spike Protein-ACE-2 Interaction.

PURPOSE: Coronil is a tri-herbal formulation containing extracts from Withania somnifera, Tinospora cordifolia, and Ocimum sanctum. Recently, it was shown that Coronil rescued humanized zebrafish from SARS-CoV-2 induced pathologies. Based on reported computational studies on the phytochemicals present in Coronil, it could be a potential inhibitor of SARS-CoV-2 entry into the host cell and associated cytokines' production. METHODS: Through an ELISA-based biochemical assay, effects of Coronil on interaction between ACE-2 and different mutants of viral spike (S) protein, crucial for viral invasion of host cell, were evaluated. Additionally, using recombinant pseudoviruses having SARS-CoV-2 spike (S) protein in their envelopes and firefly luciferase reporter in their genomes, effects of Coronil on virus entry into human alveolar epithelial cells were evaluated through luciferase assay. UHPLC profiled Coronil also modulated S-protein mediated production of pro-inflammatory cytokines in A549 cells, like interleukin-6 (IL-6), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α), as evaluated through RT-qPCR and ELISA. RESULTS: Coronil effectively inhibited the interaction of ACE-2 not only with the wild-type S protein (SWT) but also with its currently prevalent and more infectious variant (SD614G) and another mutant (SW436R) with significantly higher affinity toward ACE-2. Treatment with Coronil significantly reduced the increased levels of IL-6, IL-1ß, and TNF-α in A549 cells incubated with different S-protein variants in a dose-dependent manner. Likewise, it also prevented the SARS-CoV-2 S-protein pseudotyped vesicular stomatitis virus (VSVppSARS-2S) mediated cytokine response in these cells by reducing entry of pseudoviruses into host cells. CONCLUSION: Coronil prevented SARS-CoV-2 S-protein mediated viral entry into A549 cells by inhibiting spike protein-ACE-2 interactions. SARS-CoV-2 S protein induced inflammatory cytokine response in these cells was also moderated by Coronil.

Randomized placebo-controlled pilot clinical trial on the efficacy of ayurvedic treatment regime on COVID-19 positive patients.

BACKGROUND: Specific treatment for COVID-19 is still an unmet need. Outcomes of clinical trials on repurposed drugs have not been yielding success. Therefore, it is necessary to include complementary approaches of medicine against COVID-19. PURPOSE: This study was designed to evaluate the impact of traditional Indian Ayurvedic treatment regime on asymptomatic patients with COVID-19 infection. STUDY DESIGN: It is a placebo controlled randomized double-blind pilot clinical trial. METHODS: The study was registered with Clinical Trial Registry-India (vide Registration No. CTRI/2020/05/025273) and conducted at the Department of Medicine in National Institute of Medical Sciences and Research, Jaipur, India. 1 g of Giloy Ghanvati (Tinospora cordifolia) and 2 g of Swasari Ras (traditional herbo-mineral formulation) and 0.5 g each of Ashwagandha (Withania somnifera) and Tulsi Ghanvati (Ocimum sanctum) were given orally to the patients in treatment group twice per day for 7 days. Medicines were given in the form of tablets and each tablet weighed 500 mg. While, Swasari Ras was administered in powdered form, 30 min before breakfasts and dinners, rest were scheduled for 30 min post-meals. Patients in the treatment group also received 4 drops of Anu taila (traditional nasal drop) in each nostril every day 1 h before breakfast. Patients in the placebo group received identical-looking tablets and drops, post randomization and double blinded assortments. RT-qPCR test was used for the detection of viral load in the nasopharyngeal and oropharyngeal swab samples of study participants during the study. Chemiluminescent immunometric assay was used to quantify serum levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and high sensitivity C-reactive protein (hs-CRP) on day 1 and day 7 of the study. RESULTS: By day 3, 71.1 % and 50.0 % patients recovered in the treatment and placebo groups, respectively. Treatment group witnessed 100 % recovery by day 7, while it was 60.0 % in the placebo group. Average fold changes in serum levels of hs-CRP, IL-6 and TNF-α in treatment group were respectively, 12.4, 2.5 and 20 times lesser than those in the placebo group at day 7. There was 40 % absolute reduction in the risk of delayed recovery from infection in the treatment group. CONCLUSIONS: Ayurvedic treatment can expedite virological clearance, help in faster recovery and concomitantly reduce the risk of viral dissemination. Reduced inflammation markers suggested less severity of SARS-CoV-2 infection in the treatment group. Moreover, there was no adverse effect observed to be associated with this treatment.

Calcio-Herbal Medicine Divya-Swasari-Vati Ameliorates SARS-CoV-2 Spike Protein-Induced Pathological Features and Inflammation in Humanized Zebrafish Model by Moderating IL-6 and TNF-α Cytokines.

PURPOSE: Severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection has grown into a pandemic and without a specific cure, disease management is the need of the hour through symptomatic interventions. Studies with severe acute respiratory syndrome-coronavirus (SARS-CoV) have highlighted the role of herbal medicines either in combination with antiviral drugs or by themselves in curtailing the severity of infection and associated inflammation. Divya-Swasari-Vati is an Indian ayurvedic formulation used in the treatment of chronic cough and lung inflammation, which is one of the first symptoms of SARS-CoV-2 infections. METHODS: In this study, we used a A549 cell xenotransplant in the swim bladder of zebrafish and modeled the SARS-CoV-2 infection by injecting the fish with a recombinant spike protein. The different groups were given normal feed or feed mixed with either dexamethasone (as the control drug) or Divya-Swasari-Vati. The changes in behavioral fever, infiltration of pro-inflammatory cells in the swim bladder, degeneration or presence of necrotic cells in the kidney, and gene expression of pro-inflammatory cytokines were studied to determine the rescue of the diseased phenotype. RESULTS: Challenge with the spike protein caused changes in the swim bladder cytology with infiltrating pro-inflammatory cells, skin hemorrhage, and increase in behavioral fever. This was also accompanied by increased mortality of the disease control fish. Treatment with Divya-Swasari-Vati reversed most of the disease symptoms including damage to the kidney glomerulocytes, and complete reversal of behavioral fever. Dexamethasone, used as a comparator, was only able to partly rescue the behavioral fever phenotype. Divya-Swasari-Vati also suppressed the pro-inflammatory cytokines, IL-6 and TNF-α, levels in a dose-dependent manner, under in vivo and in vitro conditions. CONCLUSION: The study showed that the A549 xenotransplanted zebrafish injected with the recombinant spike protein of SARS-CoV-2 is an efficient model for the disease; and treatment with Divya-Swasari-Vati medicine rescued most of the inflammatory damage caused by the viral spike protein while increasing survival of the experimental fish.

GermiX: A skin friendly hand sanitizer with prolonged effectivity against pathogenic bacteria.

COVID-19 pandemic has almost made hand sanitization a ritual resulting in a steep increase in the frequency of hand sanitization and an unprecedented surge in demand for hand sanitizers. In fact, several governments had to ration hand sanitizers in the retail outlets and over the counter chemist shops. Additionally, Indian government has put a cap on the prices of hand sanitizers. Currently, large sections of global and Indian population are grappling under financial crises. Therefore, mandatory hand sanitization has made an unwelcoming, yet unavoidable addition to the already-hard-to-maintain-grocery-list. Here, we have compared the anti-microbial efficacy of Patanjali Hand Sanitizer (PHS), developed and marketed by Patanjali Ayurved Ltd. (an India-based food and herbal medicine company) with one of the topmost hand sanitizers currently used under clinical set-ups. PHS has anti-microbial efficacy comparable to that of the standard hand sanitizer. Besides, disc diffusion and time-dependent thumb print assays showed that PHS has longer retentivity on the applied surfaces, suggesting lesser consumption of the sanitizer and concomitant relaxation on the monthly grocery budget. Observed anti-bacterial potency of PHS is attributed to the disruption of bacterial cell membrane, as employed by alcohol-based hand sanitizers. A rough estimation revealed that PHS is ~ 4.3 times cost effective than the standard hand sanitizer used as the positive control in this study. Taken together, PHS is a suitable alternative for existing hand sanitizers available in the market that can relax the demand-supply strain and soften significantly the burden of monthly expenditure on hand sanitizers.

Computational Evidences of Phytochemical Mediated Disruption of PLpro Driven Replication of SARS-CoV-2: A Therapeutic Approach against COVID-19.

BACKGROUND: COVID-19 caused by SARS-CoV-2 has been declared as a global pandemic by WHO. Comprehensive analysis of this unprecedented outbreak may help to fight against the disease and may play a pivotal role in decreasing the mortality rate linked with it. Papain-like protease (PLpro), a multifunctional polyprotein, facilitates the replication of SARS-CoV-2 and evades it from the host immunological response by antagonizing cytokines, interferons and may be considered as a potential drug target to combat the current pandemic. METHODS: Natural moieties obtained from medicinal plants were analysed for their potency to target PLpro of SARS-CoV-2 by molecular docking study and were compared with synthetic analogs named as remdesivir, chloroquine and favipiravir. The stability of complexes of top hits was analysed by MD Simulation, and interaction energy was calculated. Furthermore, average RMSD values were computed and deepsite ligand-binding pockets were predicted using Play Molecule. Drug-like-abilities of these moieties were determined using ADMET and bond distance between the ligand and active site was assessed to predict the strength of the interaction. RESULTS: Nimbocinol (-7.6 Kcal/mol) and sage (-7.3 Kcal/mol) exhibited maximum BA against PLpro SARS-CoV-2 as evident from molecular docking study, which was found to be even better than remdesivir (-6.1 Kcal/mol), chloroquine (-5.3 Kcal/mol) and favipiravir (-5.7 Kcal/mol). Both nimbocinol- PLpro and sage-PLpro SARS-CoV-2 complex exhibited stable conformation during MD Simulation of 101ns at 310 K, and potential, kinetic and electrostatic interaction energies were computed, which was observed to be concordant with results of molecular docking study. RMSD average values were found to be 0.496 ± 0.015 Å and 0.598 ± 0.023 Å for nimbocinol and sage, respectively, thus revealing that both the deviation and fluctuations during MD Simulation were observed to be least. Deepsite prediction disclosed that both compounds occupied cryptic pockets in receptor and non-bond distance analysis revealed the formation of hydrogen bonds during ligand-receptor interaction. ADMET exploration further validated the drug-like properties of these compounds. CONCLUSION: Present study revealed that active constituents of Azadirachta indica and Salvia officinalis can be potentially used to target SARS-CoV-2 by hindering its replication process.