Structural interactions of phytoconstituent(s) from cinnamon, bay leaf, oregano, and parsley with SARS‐CoV‐2 nucleocapsid protein: A comparative assessment for development of potential antiviral nutraceuticals

SARS‐CoV‐2 has been responsible for causing 6,218,308 deaths globally till date and has garnered worldwide attention. The lack of effective preventive and therapeutic drugs against SARS‐CoV‐2 has further worsened the scenario and has bolstered research in the area. The N‐terminal and C‐terminal RNA binding domains (NTD and CTD) of SARS‐CoV‐2 nucleocapsid protein represent attractive therapeutic drug targets. Naturally occurring compounds are an excellent source of novel drug candidates due to their structural diversity and safety. Ten major bioactive compounds were identified in ethanolic extract (s) of Cinnamomum zeylanicum, Cinnamomum tamala, Origanum vulgare, and Petroselinum crispum using HPLC and their cytotoxic potential was determined against cancer and normal cell lines by MTT assay to ascertain their biological activity in vitro. To evaluate their antiviral potential, the binding efficacy to NTD and CTD of SARS‐CoV‐2 nucleocapsid protein was determined using in silico biology tools. In silico assessment of the phytocomponents revealed that most of the phytoconstituents displayed a druglike character with no predicted toxicity. Binding affinities were in the order apigenin > catechin > apiin toward SARS‐CoV‐2 nucleocapsid NTD. Toward nucleocapsid CTD, the affinity decreased as apigenin > cinnamic acid > catechin. Remdesivir displayed lesser affinity with NTD and CTD of SARS‐CoV‐2 nucleocapsid proteins than any of the studied phytoconstituents. Molecular dynamics (MD) simulation results revealed that throughout the 100 ns simulation, SARS‐CoV‐2 nucleocapsid protein NTD‐apigenin complex displayed greater stability than SARS‐CoV‐2 nucleocapsid protein NTD‐cinnamic acid complex. Hence, apigenin, catechin, apiin and cinnamic acid might prove as effective prophylactic and therapeutic candidates against SARS‐CoV‐2, if examined further in vitro and in vivo. PRACTICAL APPLICATIONS: Ten major bioactive compounds were identified in the extract(s) of four medicinally important plants viz. Cinnamomum zeylanicum, Cinnamomum tamala, Origanum vulgare and Petroselinum crispum using HPLC and their biological activity was also evaluated against cancer and normal cell lines. Interestingly, while all extract(s) wielded significant cytotoxicity against cancer cells, no significant toxicity was found against normal cells. The outcome of the results prompted evaluation of the antiviral potential of the ten bioactive compounds using in silico biology tools. The present study emphasizes on the application of computational approaches to understand the binding interaction and efficacy of the ten bioactive compounds from the above plants with SARS‐CoV‐2 nucleocapsid protein N‐terminal and C‐terminal RNA binding domains in preventing and/or treating COVID‐19 using in silico tools. Druglikeness and toxicity profiles of the compounds were carried out to check the therapeutic application of the components. Additionally, molecular dynamics (MD) simulation was performed to check the stability of ligand‐protein complexes. The results provided useful insights into the structural binding interaction(s) that can be exploited for the further development of potential antiviral agents targeting SARS‐CoV‐2 especially since no specific therapy is still available to combat the rapidly evolving virus and the existing treatment is more or less symptomatic which makes search for novel antiviral agents all the more necessary and crucial.

Prophylactic and therapeutic potential of selected immunomodulatory agents from Ayurveda against coronaviruses amidst the current formidable scenario: an in silico analysis.

There is currently a dearth of specific therapies to treat respiratory infections caused by the three related species of coronaviruses viz. SARS-CoV-2, SARS-CoV and MERS-CoV. Prevention from disease is currently the safest and most convenient alternative available. The present study aimed to evaluate the preventive and therapeutic effect of fifteen phytoconstituents from medicinal plants of Ayurveda against coronaviruses by in silico screening. All the phytoconstituents exhibited rapid GI absorption and bioavailability and most of them had no toxicity versus reference drug chloroquine. BAS analyses revealed that most of the phytocomponents had favorable bioactivity scores towards biological target proteins. Principal component analysis revealed that most of the phytoconstituents fell close to chloroquine in 3D projection of chemical space. Affinity of phytoconstituents towards SARS-CoV-2 spike protein-human ACE2 complex decreased as isomeldenin > tinosporaside > EGCG whereas in case of unbound ACE2, the strength of binding followed the order isomeldenin > tinosporaside > ellagic acid. Towards SARS-CoV-2 main and papain-like proteases, the affinity decreased as isomeldenin > EGCG > tinosporaside and EGCG > tinosporaside > isomeldenin, respectively. Most phytoconstituents displayed significant binding kinetics to the selected protein targets than chloroquine. SAR analysis revealed that isomeldenin, tinosporaside, EGCG and ellagic acid bind to viral spike glycoproteins via H-bond, Pi-Pi, Pi-sigma and Pi-alkyl type interactions. Molecular dynamics simulation of isomeldenin and EGCG with SARS-CoV and SARS-CoV-2 spike glycoproteins exhibited low deviations throughout the 100 ns simulation indicating good stability and compactness of the protein-ligand complexes. Thus, the above four phytoconstituents have the potential to emerge as prophylactic and therapeutic agents against coronaviruses if investigated further in vitro and in vivo. Communicated by Ramaswamy H. Sarma.

Modeling structure-activity relationships with machine learning to identify GSK3-targeted small molecules as potential COVID-19 therapeutics.

Coronaviruses induce severe upper respiratory tract infections, which can spread to the lungs. The nucleocapsid protein (N protein) plays an important role in genome replication, transcription, and virion assembly in SARS-CoV-2, the virus causing COVID-19, and in other coronaviruses. Glycogen synthase kinase 3 (GSK3) activation phosphorylates the viral N protein. To combat COVID-19 and future coronavirus outbreaks, interference with the dependence of N protein on GSK3 may be a viable strategy. Toward this end, this study aimed to construct robust machine learning models to identify GSK3 inhibitors from Food and Drug Administration-approved and investigational drug libraries using the quantitative structure-activity relationship approach. A non-redundant dataset consisting of 495 and 3070 compounds for GSK3α and GSK3ß, respectively, was acquired from the ChEMBL database. Twelve sets of molecular descriptors were used to define these inhibitors, and machine learning algorithms were selected using the LazyPredict package. Histogram-based gradient boosting and light gradient boosting machine algorithms were used to develop predictive models that were evaluated based on the root mean square error and R-squared value. Finally, the top two drugs (selinexor and ruboxistaurin) were selected for molecular dynamics simulation based on the highest predicted activity (negative log of the half-maximal inhibitory concentration, pIC50 value) to further investigate the structural stability of the protein-ligand complexes. This artificial intelligence-based virtual high-throughput screening approach is an effective strategy for accelerating drug discovery and finding novel pharmacological targets while reducing the cost and time.

Identifying How E-Service Quality Affects Perceived Usefulness of Online Reviews in Post-COVID-19 Context: A Sustainable Food Consumption Behavior Paradigm

In this study, the concept of online food purchasing is explored where consumers are not required to visit markets to purchase their foods, especially during the COVID-19 pandemic. Thus, the purpose of this study is to investigate the relationship between perceived e-service quality and related customer service outcomes in online shopping context. The influence of sustainable marketing practices in terms of perceived e-service quality (ESQ), perceived usefulness of online reviews (PUO), brand self-connection (BSC), personal innovativeness (PRI), and willingness to pay for online food services (WPO) has largely been neglected in the previous studies. The present study proposes a conceptual model to fill this gap and empirically examines how ESQ affects PUO, BSC, WPO, and e-word-of-mouth (e-WOM) of food delivery service brands. An online questionnaire survey was conducted with 423 customers utilizing the PLS-SEM-based approach to determine product indicators. Empirical results reveal that ESQ significantly influence BSC and PUO. In the same vein, PUO significantly influence BSC, while BSC significantly influence e-WOM and WPO. The results further indicate that PRI moderates the relationship between ESQ and BSC. In a post-pandemic context, our analysis indicates enormous implications for food service delivery brands in emerging economies. Online food service providers should consider reviewers' opinions about the products and services they offer and encourage their customers to write positive reviews of the products and services they offer.

Data-driven analysis and predictive modeling on COVID-19.

The coronavirus (COVID-19) started in China in 2019, has spread rapidly in every single country and has spread in millions of cases worldwide. This paper presents a proposed approach that involves identifying the relative impact of COVID-19 on a specific gender, the mortality rate in specific age, investigating different safety measures adopted by each country and their impact on the virus growth rate. Our study proposes data-driven analysis and prediction modeling by investigating three aspects of the pandemic (gender of patients, global growth rate, and social distancing). Several machine learning and ensemble models have been used and compared to obtain the best accuracy. Experiments have been demonstrated on three large public datasets. The motivation of this study is to propose an analytical machine learning based model to explore three significant aspects of COVID-19 pandemic as gender, global growth rate, and social distancing. The proposed analytical model includes classic classifiers, distinctive ensemble methods such as bagging, feature based ensemble, voting and stacking. The results show a superior prediction performance comparing with the related approaches.

Therapeutic Targeting of Innate Immune Receptors Against SARS-CoV-2 Infection.

The innate immune system is the first line of host's defense against invading pathogens. Multiple cellular sensors that detect viral components can induce innate antiviral immune responses. As a result, interferons and pro-inflammatory cytokines are produced which help in the elimination of invading viruses. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) belongs to Coronaviridae family, and has a single-stranded, positive-sense RNA genome. It can infect multiple hosts; in humans, it is responsible for the novel coronavirus disease 2019 (COVID-19). Successful, timely, and appropriate detection of SARS-CoV-2 can be very important for the early generation of the immune response. Several drugs that target the innate immune receptors as well as other signaling molecules generated during the innate immune response are currently being investigated in clinical trials. In this review, we summarized the current knowledge of the mechanisms underlying host sensing and innate immune responses against SARS-CoV-2 infection, as well as the role of innate immune receptors in terms of their therapeutic potential against SARS-CoV-2. Moreover, we discussed the drugs undergoing clinical trials and the FDA approved drugs against SARS-CoV-2. This review will help in understanding the interactions between SARS-CoV-2 and innate immune receptors and thus will point towards new dimensions for the development of new therapeutics, which can be beneficial in the current pandemic.

Intelligent Facemask Coverage Detector in a World of Chaos

The recent outbreak of COVID-19 around the world has caused a global health catastrophe along with economic consequences. As per the World Health Organization (WHO), this devastating crisis can be minimized and controlled if humans wear facemasks in public;however, the prevention of spreading COVID-19 can only be possible only if they are worn properly, covering both the nose and mouth. Nonetheless, in public places or in chaos, a manual check of persons wearing the masks properly or not is a hectic job and can cause panic. For such conditions, an automatic mask-wearing system is desired. Therefore, this study analyzed several deep learning pre-trained networks and classical machine learning algorithms that can automatically detect whether the person wears the facemask or not. For this, 40,000 images are utilized to train and test 9 different models, namely, InceptionV3, EfficientNetB0, EfficientNetB2, DenseNet201, ResNet152, VGG19, convolutional neural network (CNN), support vector machine (SVM), and random forest (RF), to recognize facemasks in images. Besides just detecting the mask, the trained models also detect whether the person is wearing the mask properly (covering nose and mouth), partially (mouth only), or wearing it inappropriately (not covering nose and mouth). Experimental work reveals that InceptionV3 and EfficientNetB2 outperformed all other methods by attaining an overall accuracy of around 98.40% and a precision, recall, and F1-score of 98.30%.

Therapeutic significance of nano- and biosensor technology in combating SARS-CoV-2: a review.

The diagnosis of novel coronavirus (COVID-19) has gained the spotlight of the world's scientific community since December 2019 and it remains an important issue due to the emergence of novel variants around the globe. Early diagnosis of coronavirus is captious to prevent and hard to control. This pandemic can be eradicated by implementing suppressing strategies which can lead to better outcomes and more lives being saved. Therefore, the analysis showed that COVID-19 can only be managed by adopting public health measures, such as testing, isolation and social distancing. Much work has been done to diagnose coronavirus. Various testing technologies have been developed, opted and modified for rapid and accurate detection. The advanced molecular diagnosis relies on the detection of SARS-CoV-2 as it has been considered the main causative agent of this pandemic. Studies have shown that several molecular tests are considered essential for the confirmation of coronavirus infection. Various serology-based tests are also used in the detection and diagnosis of coronavirus including point-of-care assays and high-throughput enzyme immunoassays that aid in the diagnosis of COVID-19. Both these assays are time-consuming and have less diagnostic accuracy. Nanotechnology has the potential to develop new strategies to combat COVID-19 by developing diagnostics and therapeutics. In this review, we have focused on the nanotechnology-based detection techniques including nanoparticles and biosensors to obstruct the spread of SARS-CoV-2.

Therapeutic Targeting of Innate Immune Receptors Against SARS-CoV-2 Infection

The innate immune system is the first line of host’s defense against invading pathogens. Multiple cellular sensors that detect viral components can induce innate antiviral immune responses. As a result, interferons and pro-inflammatory cytokines are produced which help in the elimination of invading viruses. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) belongs to Coronaviridae family, and has a single-stranded, positive-sense RNA genome. It can infect multiple hosts;in humans, it is responsible for the novel coronavirus disease 2019 (COVID-19). Successful, timely, and appropriate detection of SARS-CoV-2 can be very important for the early generation of the immune response. Several drugs that target the innate immune receptors as well as other signaling molecules generated during the innate immune response are currently being investigated in clinical trials. In this review, we summarized the current knowledge of the mechanisms underlying host sensing and innate immune responses against SARS-CoV-2 infection, as well as the role of innate immune receptors in terms of their therapeutic potential against SARS-CoV-2. Moreover, we discussed the drugs undergoing clinical trials and the FDA approved drugs against SARS-CoV-2. This review will help in understanding the interactions between SARS-CoV-2 and innate immune receptors and thus will point towards new dimensions for the development of new therapeutics, which can be beneficial in the current pandemic.

Structural interactions of phytoconstituent(s) from cinnamon, bay leaf, oregano, and parsley with SARS-CoV-2 nucleocapsid protein: A comparative assessment for development of potential antiviral nutraceuticals.

SARS-CoV-2 has been responsible for causing 6,218,308 deaths globally till date and has garnered worldwide attention. The lack of effective preventive and therapeutic drugs against SARS-CoV-2 has further worsened the scenario and has bolstered research in the area. The N-terminal and C-terminal RNA binding domains (NTD and CTD) of SARS-CoV-2 nucleocapsid protein represent attractive therapeutic drug targets. Naturally occurring compounds are an excellent source of novel drug candidates due to their structural diversity and safety. Ten major bioactive compounds were identified in ethanolic extract (s) of Cinnamomum zeylanicum, Cinnamomum tamala, Origanum vulgare, and Petroselinum crispum using HPLC and their cytotoxic potential was determined against cancer and normal cell lines by MTT assay to ascertain their biological activity in vitro. To evaluate their antiviral potential, the binding efficacy to NTD and CTD of SARS-CoV-2 nucleocapsid protein was determined using in silico biology tools. In silico assessment of the phytocomponents revealed that most of the phytoconstituents displayed a druglike character with no predicted toxicity. Binding affinities were in the order apigenin > catechin > apiin toward SARS-CoV-2 nucleocapsid NTD. Toward nucleocapsid CTD, the affinity decreased as apigenin > cinnamic acid > catechin. Remdesivir displayed lesser affinity with NTD and CTD of SARS-CoV-2 nucleocapsid proteins than any of the studied phytoconstituents. Molecular dynamics (MD) simulation results revealed that throughout the 100 ns simulation, SARS-CoV-2 nucleocapsid protein NTD-apigenin complex displayed greater stability than SARS-CoV-2 nucleocapsid protein NTD-cinnamic acid complex. Hence, apigenin, catechin, apiin and cinnamic acid might prove as effective prophylactic and therapeutic candidates against SARS-CoV-2, if examined further in vitro and in vivo. PRACTICAL APPLICATIONS: Ten major bioactive compounds were identified in the extract(s) of four medicinally important plants viz. Cinnamomum zeylanicum, Cinnamomum tamala, Origanum vulgare and Petroselinum crispum using HPLC and their biological activity was also evaluated against cancer and normal cell lines. Interestingly, while all extract(s) wielded significant cytotoxicity against cancer cells, no significant toxicity was found against normal cells. The outcome of the results prompted evaluation of the antiviral potential of the ten bioactive compounds using in silico biology tools. The present study emphasizes on the application of computational approaches to understand the binding interaction and efficacy of the ten bioactive compounds from the above plants with SARS-CoV-2 nucleocapsid protein N-terminal and C-terminal RNA binding domains in preventing and/or treating COVID-19 using in silico tools. Druglikeness and toxicity profiles of the compounds were carried out to check the therapeutic application of the components. Additionally, molecular dynamics (MD) simulation was performed to check the stability of ligand-protein complexes. The results provided useful insights into the structural binding interaction(s) that can be exploited for the further development of potential antiviral agents targeting SARS-CoV-2 especially since no specific therapy is still available to combat the rapidly evolving virus and the existing treatment is more or less symptomatic which makes search for novel antiviral agents all the more necessary and crucial.

Ethno pharmacology, Phytochemical Analysis, Safety Profile, Prophylactic Aspects, and Therapeutic Potential of Asarum europaeum L. in Unani Medicine: An Evidence-Based Appraisal

Background : A. europaeum is a well mentioned Unani herb, belonging to Aristolochiaceae family and indigenous to the Eastern and Southern Europe. A. europaeum has traditionally been utilised in Unani medicine to treat respiratory ailments, gastrointestinal disorders, neurological disorders, hepatic disorders, genitourinary disorders, snake poisoning, vector-borne infections, and epidemic prophylaxis. This review aims to the compile previous and present available information concerning about the ethno-botanical aspects, phyto-chemical constituents, isolated metabolites, and pharmacological profiling of this herb correlating the principles of Unani doctrine, thereby may offer series of valuable data for researchers and pharmaceuticals to develop novel therapeutic ways. ‏ Material and methods : A detailed literature review has been conducted on A. europaeum in various electronic databases, including Pub Med, Web of Science, Wiley, Science Direct, Elsevier, Google Scholar, ACS publications, Springer Link etc. Furthermore, books (Unani ancient classical books) were consulted in Urdu, Arabic, Persian and English to collate the particulars. Results : The present literature survey incorrigibly illustrated the usage of A. europaeum as a prophylactic drug in several diseases such as chicken pox, plague, and viral fevers, including COVID-19 and therapeutically, aid in managing bronchitis, epilepsy, hepatitis, ascites, amenorrhea, paralysis, sciatica, lumbago, and urolithiasis. It has been well documented its usage in the form of powder, decoction, syrup, extract, and oil as a single drug or in compound formulations with the amalgamation of other herbs. Phyto-chemical analysis from various sections of the plant revealed the presence of varied chemical constituents for example essential oils (with four chemotypes), flavonoids, phenolic compounds, organic acids, vitamins, terpenes and sesquiterpenes. It has also been evaluated through in vitro, in vivo, in silico, preclinical and clinical trial models for diverse pharmacological activities such as antibacterial, anti-diabetic, and anti-proliferative activities, etc. Still, only sporadic herb studies have been published so far. Conclusion : The current paper highlights botany, ethno pharmacology, phyto-chemistry, pharmacology, and toxicology in depth. Preliminary pharmacological studies on different extracts and fractions of A. europaeum support the claim of Unani scholars that it is useful in the treatment of various diseases. However, immediate efforts must be made to determine its mechanism of action, efficacy, dosage, and safety in combating various pathological states. The study will undoubtedly serve as the basis for future research to further demonstrate the ethno medicinal and therapeutic potential for health-care product improvement.

Effect of Chloroquine and Hydroxychloroquine on Cytokine Release Syndrome in Patients with COVID-19.

Objective: To assess the effect of chloroquine and hydroxychloroquine on cytokine release syndrome (CRS) in adult patients with coronavirus disease 2019 (COVID-19) having mild to moderate symptoms.Methods: This blinded, placebo-controlled, randomized study was conducted in the Department of Medicine, Pak Emirates Military Hospital Rawalpindi, from June 1-15, 2020. A total of 150 hospitalized patients were enrolled after diagnoses with COVID-19 through reverse transcription polymerase chain reaction (RT-PCR). They were divided into three groups: hydroxychloroquine plus general care (HGC, n=50), chloroquine plus general care (CGC, n=50); and only general care (OGC, n=50). The HGC group received treatment with hydroxychloroquine 400 mg every 12 hours for day one and 200 mg for the next 4 days. The CGC group received treatment with chloroquine 250 mg every 12 hours for 7 days. The OGC group was kept as a control with only general care. After 12 days, the patients were screened for development of CRS through detection of interleukin 6 (IL-6) in serum samples by using Roche cobas e411 electrochemiluminescence immunoassay analyzer.Results: The mean duration from onset of symptoms to randomization was 7.65 days (SD = 3.287 days; range, 2-15 days). The mean age of patients was 37.57 (range 19-63) years. Results showed that out of a total 150 patients, only 10 patients (6%, mean=1.93; CI=1.89-1.97, P=0.651) developed CRS in all study groups. Four patients (8%) developed CRS in the HGC group, 2 patients (4%) in the CGC group, and 4 patients (8%) in the OGC group. There was no significant difference in the mean level of CRS among study groups.Conclusion: Administration of hydroxychloroquine and chloroquine has no effect in reducing the development of CRS in patients with COVID-19 having mild to moderate symptoms.

Early Fall in C-Reactive Protein (CRP) Level Predicts Response to Tocilizumab in Rapidly Progressing COVID-19: Experience in a Single-Arm Pakistani Center.

Introduction There are conflicting studies regarding the efficacy of tocilizumab use in coronavirus disease 2019 (COVID-19) disease. There is a special need to identify the parameters that could predict its response in early COVID-19 disease. Objective To report our experience with tocilizumab and correlate the magnitude of fall in c-reactive protein (CRP) as a predictor of its response to treatment in early COVID-19 disease. Methods All confirmed COVID-19 cases admitted to a tertiary healthcare hospital in Peshawar Pakistan, receiving ≥1 dose of intravenous tocilizumab, between March and September 2020 were included. Relevant clinical data of the patients were recorded and further divided into two categories based on the relative fall in CRP levels, 48 hours after tocilizumab administration. Adequate response (≥50% fall from baseline CRP), primary outcomes (fall in oxygen requirement and inflammatory biomarkers), and secondary outcome (all-cause mortality at day 28) were recorded. All outcomes were compared based on falls in CRP levels. Results A total of 27 patients were included. Males were 24 (88.8%) while females were three (11.1%). The mean age was 60.9±11.6 years. The mean day of illness at the time of tocilizumab administration was 4.26±3 days. After 48 hours of tocilizumab administration, 17 (62.9%) patients showed clinical improvement, with the mean SaO2/FiO2 ratio prior to treatment significantly increased (p<0.01). A significant reduction in CRP and ferritin levels was seen post-treatment (p <0.01 and p<0.01, respectively). Twenty (74.1%) patients demonstrated adequate response to tocilizumab while seven (25.9%) showed an inadequate response. Patients with adequate response had higher chances of improvement in oxygenation and lower in-hospital mortality (p-value 0.009 and 0.020, respectively). Conclusions Tocilizumab shows clinical improvement in a vast majority of patients. Being an early and sensitive predictor, a fall of ≥50% in CRP at 48 hours can be used to predict the overall response to tocilizumab as a guide to treatment.

Dermatological Manifestations of COVID-19 in Patients Reporting to a Tertiary Care Hospital in Rawalpindi, Pakistan.

Introduction The clinicopathological description of dermatological manifestations of COVID-19 leaves much to be desired. There is a need to determine their association with disease severity, outcome, and other clinical variables. Objectives The objectives of this study are to record and histopathologically examine the cutaneous manifestations of COVID-19 and correlate these to age, disease severity, and mortality. Methods All confirmed COVID-19 patients admitted to a single tertiary healthcare hospital in Rawalpindi, Pakistan, were included. Their diseases were classified as mild, moderate, severe, and critical. The recent onset skin eruptions in these patients were recorded via photographs along with relevant clinical data. The photographs were independently reviewed by a group of three dermatologists without knowledge of the clinical information. The skin manifestations were divided into disease-specific and nonspecific categories using an already defined algorithm. Histopathological examination of skin manifestations was conducted. Results A total of 23% (n=47) had "new" skin manifestations. Specific skin findings were seen in 21.6% (n=44), which consisted of ecchymosis/purpura in 50% (n=22), maculopapular exanthem in 18% (n=8), livedo reticularis in 16.2% (n=7), ischemia/gangrene in 16.2% (n=7), perniosis in 15.9 % (n=7), vesiculo-bullous rash in 9% (n=4) and urticaria in 4% (n=1). Non-specific findings were seen in 6% (n=13) and included bedsores, dermatitis passivata, dryness, herpes labialis, oral ulcerations, and nasogastric tube-induced ulcerations. There was a significant association (p=0.03) between disease severity and specific skin lesions. Ischemia/gangrene was significantly associated with COVID-19 disease severity and mortality. Vesiculobullous lesions were associated with higher mortality, though not with disease severity. Livedo reticularis had a higher-than-expected count in critical disease, albeit statistically insignificant. The association of maculopapular exanthem and ecchymosis/purpura with severe/critical disease was statistically insignificant. Urticaria was significantly associated with low disease severity. Mean age with specific manifestations was 56.86 ± 15.81 and with nonspecific/without any manifestations was 42.58 ± 16.96, a highly significant difference, with p-value < 0.001. Old age (>60 years) was significantly associated with ecchymosis (p=0.038), maculopapular exanthem (p=0.021), and vesiculo-bullous rash (p=0.029). Histopathology varied according to the type of skin lesion. Conclusions Dermatological manifestations coexist in many patients and tend to appear more in severe cases of COVID-19 among the older age group and only minimally in mild/moderate cases. Their presence could help set prognostic criteria of COVID-19 disease in the future.

Methylprednisolone for COVID-19 Patients Admitted to a Tertiary Care Hospital: A Single-Centre Study.

BACKGROUND: The role of various corticosteroids in the management of COVID-19 is evolving. Following an initial lack of evidence, the relatively novel data, supporting the survival benefit to severe and critical COVID-19 patients, is of limited scale. MATERIALS AND METHODS: This retrospective study observed medical records and disease outcomes of 200 patients with moderate, severe and critical COVID-19 receiving methylprednisolone (MP). The dose of methylprednisolone was 0.5 to 2 mg per kg in these patients. RESULTS: Median age of presentation was 59 years. The median duration of symptoms at presentation was five days. The most common presenting symptoms were cough (77.5%), fever (67.5%) and shortness of breath (63.5%). Majority of patients (85%) presented in the first week of illness. One or more comorbidities were present in 75% of patients. Complications seen in the study cohort were cytokine release syndrome (CRS) 92 (46%), acute respiratory distress syndrome (ARDS) 44 (22%) and multi-organ dysfunction 17 (8.5%). The median time for initiation of corticosteroid therapy was four hours. Overall survival (OS) in patients receiving methylprednisolone was 83.5%. The OS for patients with moderate, severe and critical diseases was 97.8%, 86.2% and 62%, respectively (p<0.001). CONCLUSION: Steroids like methylprednisolone are useful in COVID-19 admitted patients and provide excellent survival outcomes.

Cognitive behavioral crisis intervention model for health care professionals in Pakistan

The health care professionals (HCPs) in Pakistan are vulnerable to the negative psychological impact of the current COVID - 19 pandemic. The shortage of trained HCPs in Pakistan coupled with vulnerable infrastructure and depleted resources make the situation a source of psychological reactions like fear, anger, anxiety, and depression for HCPs during COVID - 19 outbreak. These psychological reactions are produced by the preceding thoughts and emotions according to the cognitive behavioral model. Therefore, a cognitive behavioral crisis intervention model (CBCIM) is proposed with aim of helping HCPs deal with these psychological reactions efficiently. The common components of CBCIM include the cognitive restructuring of the negative thoughts, teaching of relaxation and mindfulness exercises, the teaching of ACT - ADD approach and use of coping cards, district wise team - based act ion plan and the provision of these services to HCPs regularly even after the end of COVID - 19 pandemic.

Exploring the Binding Mechanism of PF-07321332 SARS-CoV-2 Protease Inhibitor through Molecular Dynamics and Binding Free Energy Simulations.

The novel coronavirus disease, caused by severe acute respiratory coronavirus 2 (SARS-CoV-2), rapidly spreading around the world, poses a major threat to the global public health. Herein, we demonstrated the binding mechanism of PF-07321332, α-ketoamide, lopinavir, and ritonavir to the coronavirus 3-chymotrypsin-like-protease (3CLpro) by means of docking and molecular dynamic (MD) simulations. The analysis of MD trajectories of 3CLpro with PF-07321332, α-ketoamide, lopinavir, and ritonavir revealed that 3CLpro-PF-07321332 and 3CLpro-α-ketoamide complexes remained stable compared with 3CLpro-ritonavir and 3CLpro-lopinavir. Investigating the dynamic behavior of ligand-protein interaction, ligands PF-07321332 and α-ketoamide showed stronger bonding via making interactions with catalytic dyad residues His41-Cys145 of 3CLpro. Lopinavir and ritonavir were unable to disrupt the catalytic dyad, as illustrated by increased bond length during the MD simulation. To decipher the ligand binding mode and affinity, ligand interactions with SARS-CoV-2 proteases and binding energy were calculated. The binding energy of the bespoke antiviral PF-07321332 clinical candidate was two times higher than that of α-ketoamide and three times than that of lopinavir and ritonavir. Our study elucidated in detail the binding mechanism of the potent PF-07321332 to 3CLpro along with the low potency of lopinavir and ritonavir due to weak binding affinity demonstrated by the binding energy data. This study will be helpful for the development and optimization of more specific compounds to combat coronavirus disease.

An Evolutionary Fake News Detection Method for COVID-19 Pandemic Information

As the COVID-19 pandemic rapidly spreads across the world, regrettably, misinformation and fake news related to COVID-19 have also spread remarkably. Such misinformation has confused people. To be able to detect such COVID-19 misinformation, an effective detection method should be applied to obtain more accurate information. This will help people and researchers easily differentiate between true and fake news. The objective of this research was to introduce an enhanced evolutionary detection approach to obtain better results compared with the previous approaches. The proposed approach aimed to reduce the number of symmetrical features and obtain a high accuracy after implementing three wrapper feature selections for evolutionary classifications using particle swarm optimization (PSO), the genetic algorithm (GA), and the salp swarm algorithm (SSA). The experiments were conducted on one of the popular datasets called the Koirala dataset. Based on the obtained prediction results, the proposed model revealed an optimistic and superior predictability performance with a high accuracy (75.4%) and reduced the number of features to 303. In addition, by comparison with other state-of-the-art classifiers, our results showed that the proposed detection method with the genetic algorithm model outperformed other classifiers in the accuracy.

SNPs at 3'UTR of APOL1 and miR-6741-3p target sites associated with kidney diseases more susceptible to SARS-COV-2 infection: in silco and in vitro studies.

Acute Kidney Injury (AKI) is a common manifestation of COVID-19 and several cases have been reported in the setting of the high-risk APOL1 genotype (common genetic variants). This increases the likelihood that African American people with the high-risk genotype APOL1 are at increased risk for kidney disease in the COVID-19 environment. Single-nucleotide polymorphisms (SNPs) are found in various microRNAs (miRNAs) and target genes change the miRNA activity that leads to different diseases. Evidence has shown that SNPs increase/decrease the effectiveness of the interaction between miRNAs and disease-related target genes. The aim of this study is not only to identify miRSNPs on the APOL1 gene and SNPs in miRNA genes targeting 3'UTR but also to evaluate the effect of these gene variations in kidney patients and their association with SARS-COV-2 infection. In 3'UTR of the APOL1 gene, we detected 96 miRNA binding sites and 35 different SNPs with 10 different online software in the binding sites of the miRNA (in silico). Also we studied gene expression of patients and control samples by using qRT-PCR (in vitro). In silico study, the binding site of miR-6741-3p on APOL1 has two SNPs (rs1288875001, G > C; rs1452517383, A > C) on APOL1 3'UTR, and its genomic sequence is the same nucleotide as rs1288875001. Similarly, two other SNPs (rs1142591, T > A; rs376326225, G > A) were identified in the binding sites of miR-6741-3p at the first position. Here, the miRSNP (rs1288875001) in APOL1 3'UTR and SNP (rs376326225) in the miR-6741-3p genomic sequence are cross-matched in the same binding region. In vitro study, the relative expression levels were calculated by the 2-ΔΔCt method & Mann-Whitney U test. The expression of APOL1 gene was different in chronic kidney patients along with COVID-19. By these results, APOL1 expression was found lower in patients than healthy (p < 0.05) in kidney patients along with COVID-19. In addition, miR-6741-3p targets many APOL1-related genes (TLR7, SLC6A19, IL-6,10,18, chemokine (C-C motif) ligand 5, SWT1, NFYB, BRF1, HES2, NFYB, MED12L, MAFG, GTF2H5, TRAF3, angiotensin II receptor-associated protein, PRSS23) by evaluating online software in the binding sites of the miR-6741-3p. miR-6741-3p has not previously shown any association with kidney diseases and SARS-COV-2 infection. It assures that APOL1 can have a significant consequence in kidney-associated diseases by different pathways. Henceforth, this study represents and demonstrates an effective association between miR-6741-3p and kidney diseases, i.e., collapsing glomerulopathy, chronic kidney disease (CKD), acute kidney injury (AKI), and tubulointerstitial lesions susceptibility to SARS-COV-2 infection via in silico and in vitro exploration and recommended to have better insight.

Non-Hodgkin lymphoma of bone of the femur and humerus: a case report and review of the literature.

Lymphoma of bone is a rare neoplasm composed of malignant lymphoid cells, producing a tumefactive lesion within bone. We report a 13-year-old male who presented with progressively increasing swellings at the right shoulder and right mid-thigh for one month. Radiological images revealed lytic destructive lesions associated with soft tissue masses in both sites and a pathological fracture on the right humerus. The patient had no significant medical history. Histological, immunohistochemical and fluorescent in-situ hybridization assessment of biopsies from the lesions confirmed the diagnosis of primary non-Hodgkin lymphoma of bone. Unfortunately, due to coronavirus disease 2019 outbreak, the patient was unable to follow-up treatment and died shortly after establishment of the diagnosis. Delay in diagnosis and treatment is of serious concern when it comes to improve the prognosis of patients with this disease.