Browsing by Author "Shrestha, Asmita"

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    COVID-19 Global Pandemic: Analysis of Secondary Metabolites from Natural Products to Aid the Engineering of Drugs
    (Institute of Science and Technology, 2024-08) Shrestha, Asmita; Dr. Salyan Bhattarai, Dr. Khaga Raj Sharma
    SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2), the pathogenic agent of Coronavirus (CoV) disease-19 (COVID-19), has wreaked havoc around the world since the end of 2019, underscoring the need for effective COVID-19 treatment. Even though COVID-19 is no longer a pandemic, research should still be done to find effective treatments for these potentially lethal CoVs because new and deadly CoVs can appear at any time and represent a threat of spreading new CoV pandemics. The frequent unfolding of new mutants of the virus may impose serious concerns again regarding the efficacy of therapeutic agents, therefore any advancement toward the development of potent antiviral medications is always critical. Since secondary metabolites have been used to cure several ailments, they could be useful in managing COVID-19. Quantitative understanding and characterization of dynamic molecular processes are key issues in modern biology and need sophisticated computational experimental methodologies to get molecular-level information on ligand-binding configurations, interactions, and dynamic conformational landscapes. To better understand the basis of the activity of secondary metabolites with biological activity, this research incorporated the pharmacokinetics study, molecular docking, molecular dynamics simulations, binding free energy (BFE) calculation, and density functional theory (DFT) analyses utilizing different computational tools followed by spike protein assays. The study began with the selection of a diverse library of 191 metabolites relying on their antiviral properties known for their potential bioactivities. These metabolites were screened against key viral proteins: S1-RBD of spike (S) protein and RNA-dependent-RNA-polymerase (RdRp); involved in the viral entry, replication, and infection processes of SARS-CoV-2. Based on in silico investigation, cordifolioside A was identified to be the most potent of the 26 secondary metabolites selected from Tinospora Species. It showed proper binding free energy of -25.09 kcal/mol, significant GOLD fitness score (58.27), reactivity, suitable pharmacokinetic profile along with considerable stability with target S1-RBD of SARS-CoV-2. Furthermore, the in vitro experiment using Tinospora cordifolia crude extract, which was conducted because it contains an appropriate quantity of cordifolioside A, revealed that it inhibits the binding of human angiotensin converting enzyme 2 (hACE2) with S1-RBD of SARS-CoV-2 by 50 % at a concentration of approximately 1.25 mg/mL. Of the 36 flavonoids, cyanidin was determined to be the more potent due to its adequate pharmacokinetic profile, binding free energy (-25.09 kcal/mol) with proper interactions, GOLD fitness score of 51.91, appropriate stability of its complex with S1-RBD, and sufficient reactivity. Additionally, as cyanidin is present in red grapes (Vitis vinifera), in vitro investigation is carried out on it and the findings indicate that it significantly interferes by 50 % at a concentration of approximately 1.25 mg/mL to the interaction of hACE2 with S1-RBD of SARS-CoV-2. Neferine, an alkaloid, was found to be an inhibitor of RdRp and S1-RBD of SARS-CoV-2 among screened 79 selected compounds. Neferine exhibited the appropriate stability and reactivity when complexed with RdRp, exhibiting an S-score of -5.083 and a binding energy of -9.1 kcal/mol. Neferine also demonstrated notable interactions with key amino acid residues in S1-RBD of the SARS-CoV-2 Omicron variant, with a binding energy of -7.6 kcal/mol and in vitro assay of neferine shows its acceptable binding with S1-RBD accompanied by IC50 of 99.60 ± 0.328 μg/mL. Among the 50 terpenes, cryptotanshinone was discovered to be a strong inhibitor of the S1-RBD Omicron variant, exhibiting stability, proper pharmacokinetic profile, and good binding affinity (-7.6 kcal/mol) with target protein. With an IC50 of 25.5 ± 0.463 µg/mL, cryptotanshinone was able to inhibit the binding of hACE 2 with S1-RBD of the SARS-CoV-2 Omicron variant, validating the results of the in silico analysis. In conclusion, this thesis offers a comprehensive computational study for the identification of secondary metabolites with potential inhibitory effects against SARS-CoV-2. The results present a significant contribution to the search for effective and safe treatments for COVID-19. Further preclinical investigation is necessary to validate and optimize these promising metabolites as potential COVID-19 therapeutics. कोभिड–१९ -COVID–19_ रोगको सङक्रमण सार्स–कोभ–२ -SARS–CoV–2_ नामक भाइरसको कारणले हुन्छ । जसले विश्वमा सन् २०१९ को अन्त्यबाट जटिल स्वास्थ्य समस्या सिर्जना गरेको थियो । पछिल्लो समय कोभिड–१९ को जोखिम विश्वमा कम भएता पनि यसको पुर्ण निवारण वा निर्मुल भएको अवस्था छैन । विभिन्न कारणले गर्दा सार्स–कोभ–२ मा म्युटेशन (एमिनो एसिडमा बदलाब हुने रसायनिक क्रिया) भई यसको नयाँ स्वरूपहरु -Variants_ संसारमा देखा परिरहेका छन् । भाइरसको नयाँ स्वरूपले यसबाट हुने सङक्रमणको जोखिम कायम नै राखेको छ । त्यसैले यस विषयमा थप वैज्ञानिक अनुसन्धानको आवश्यकता रहेको देखिन्छ । सार्स–कोभ–२ मा हुने यस्तो म्युटेसनले चिकित्साशास्त्रका क्षेत्रमा थप चिन्ता र चासो थपेको छ । त्यसैले भरपर्दो एन्टिभाइरल औषधी वा खोपको विकास तर्फ अझै प्रगती गर्नु पर्ने देखिएको छ । वनस्पतीबाट भाइरस नियन्त्रणको लागि औषधीजन्य गुण भएका यौगिक पत्ता लगाउनु यस अनुसन्धानको प्रमुख उदेश्य हो किनकी वैज्ञानिकहरूले गरेका यस अगाडीका शोधले पनि त्यस्तो संभावना प्रष्टाएको छ । त्यस्ता यौगिकले सार्स–कोभ–२ को प्रोटिनमा के कस्तो रसायनिक गुण देखाउछ, सोको गहन खोजमा यो अध्ययन केन्द्रित गरिएको छ । जसको लागि यस अनुसन्धानमा फार्माकोकाइनेटिक्स अध्ययन -Pharmacokinetics study_, मलिक्युलर डकिङ् -Molecular docking_, मलिक्युलर सिमुलेशन -Molecular Dynamics Simulation_, र डेन्सिटी फङसन सिद्धान्त -DFT_ मा आधारित विश्लेषण विभिन्न कम्प्यूटेशनल सफ्टवेयरको माध्यमबाट गरिएको छ । यस अनुसन्धानमा वनस्पतीबाट निकालिएका १९१ वटा यौगिकमा सार्स–कोभ–२ को स्पाइक प्रोटिन -Spike Protein_ र आरएनए–डिपेन्डेन्ट–आरएनए–पोलिमरेज -RdRp_ मा कम्प्यूटेशनल अध्ययन गरिएको छ । मलिक्युलर सिमुलेशन नामक कम्प्युटेशन अनुसन्धानले यसको थप पुष्टी पनि गरेको छ । यस्तो सैध्दान्तिक अनुसन्धानलाई थप वैज्ञानिक आधारहरू प्रदान गर्न प्रयोगशालामा आधारित प्रोटिन एस्से -Protein assay_पनि गरिएको छ । इन सिलिको -In silico_ अनुसन्धानको आधारमा, टिनोस्पोरा प्रजातिहरूबाट -Tinospora species_चयन गरिएका २६ secondary मेटाबोलाइटहरूमध्ये कर्डिफोलियोसाइड ए लाई -cordifolioside A_ सबैभन्दा शक्तिशाली मानिएको छ । यसले (-@%=)( kcal/mol) को उचित binding free energy, महत्त्वपूर्ण गोल्ड -GOLD_ फिटनेस स्कोर, प्रतिक्रियाशीलता, उपयुक्त फार्माकोकाइनेटिक प्रोफाइल -pharmacokinetic profile_ र SARS–CoV–2 को लक्ष्य S1–RBD सँग पर्याप्त स्थिरता देखाएको छ । यस बाहेक, Tinospora cordifolia कच्चा एक्स्ट्र्याक्ट प्रयोग गरी इन भिट्रो (in vitro) सञ्चालन गरिएको थियो किनभने यसमा कर्डिफोलियोसाइड ए को उपयुक्त मात्रा हुन्छ, यसले SARS–CoV–2 को S1–RBD सँग मानव एन्जियाटेन्सिन रूपान्तरण गर्ने इन्जाइम 2 (ACE2) लाई लगभग !.@% mg/mL को एकाग्रतामा ५० % ले बाइन्ड गर्न रोक्छ । ३६ फ्लेभोनोइडहरू मध्ये, cyanidin, यसको पर्याप्त फार्माकोकाइनेटिक प्रोफाइल, उचित अन्तरक्रियाको साथ binding free energy (-@%=)( kcal/mol) , ५१.९१ को गोल्ड फिटनेस स्कोर, S1–RBD संग यसको कम्प्लेक्सको उपयुक्त स्थिरता, र पर्याप्त प्रतिक्रियाशीलता। थप रूपमा, रातो अंगूर -Vitis vinifera_ मा साइनिडिन (cyanidin) पाइने भएकोले, यसले SARS–CoV–2 को S1–RBD सँग मानव एन्जियोटेन्सिन रूपान्तरण गर्ने इन्जाइम 2 (ACE2) लाई लगभग !.@% mg/mL को एकाग्रतामा ५० % ले बाइन्ड गर्न रोक्छ। स्क्रिन गरी चयन गरिएका ७९ एल्कालोइडहरूमध्ये नेफेरिन -neferine_ SARS–CoV–2 को RdRp र S1–RBD को अवरोधक भएको पाइयो । RdRp संग bind हुँदा neferine ले उपयुक्त स्थिरता र प्रतिक्रियाशीलता प्रदर्शन गर्यो, –५.०८३ को S–स्कोर र –(=! kcal/mol को binding energy प्रदर्शन गर्दै नेफेरिनले SARS–CoV–2 Omicron भेरियन्टको S1–RBD मा मुख्य एमिनो एसिड अवशेषहरूसँग (amino acid residues) –&=^ kcal/mol को binding energy को साथ उल्लेखनीय अन्तरक्रिया पनि प्रदर्शन ग¥यो र नेफेरिनको इन भिट्रो परीक्षणले S1-RBD सँग ९९.६० + ०.३२८ μg/mL को IC50 सँग स्वीकार्य बाइन्डिङ देखाउँछ । ५० ट्रपिनहरू -terpenes_ मध्ये, क्रिप्टोटानशिनोन -cryptotanshinone_ S1–RBD ओमिक्रोन भेरियन्टको बलियो अवरोधकको रूपमा पत्ता लगाइएको छ, जसले स्थिरता, उचित फार्माकोकाइनेटिक प्रोफाइल, र लक्ष्य प्रोटीनको साथ राम्रो बन्धन सम्बन्ध (–७.६ kcal/mol_ प्रदर्शन गर्दछ । इन सिलिको विश्लेषणको नतिजालाई प्रमाणित गर्दै २५.५ + ०.४६३ µg/mL को IC50 सँग, क्रिप्टोटान्सिनोनले SARS–CoV–2 Omicron भेरियन्टको S1–RBD सँग hACE 2 को बाइन्डिङ्लाई रोक्न सक्षम छ । यस अनुसन्धानले कोभिड–१९ को औषधी विकास गर्न प्रभावकारी र सुरक्षित उपचारहरूको खोजीमा महत्वपूर्ण योगदान प्रस्तुत गरेको छ ।
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    Maternal and Child Health Care Practice Among Newar and Non-Newar Women in Jhaukhel VDC Of Bhaktapur District
    (Department of Health Education, 2012) Shrestha, Asmita
    The study entitled “Maternal and Child Health Care Practice among Newar and Non-newar Women in Jhaukhel VDC of Bhaktapur District” was carried out in order to identify the practice of maternal and child health care. The study was mainly based on primary data collected from 2 wards (5 & 7). The married women were interviewed to collect the information. The main objectives of the study were to find out the socio-economic & demographic characteristics and the MCH care practice of the Jhaukhel VDC. The study was based upon the data collected from primary sources among 100 households (43 Newar and 57 Non-newar). The researcher used the interview schedule to collect the data. The collected data were tabulated for analysis and descriptive method was used to analyze and interpret the tables. According to this study, most of the Newar respondents (74.42%) were more educated than Non-newar respondents (71.93%). The ANC visit was satisfactory. Most of the respondents had taken the iron tablet, however they were not regular visit. 51 percent of Newar and 49 percent Non-newar respondents had received the TT vaccine and iron tablet. About 80 percent Newar respondents had given birth at hospital whereas 81 percent Non-newar respondents had given birth at hospital which is favorable aspect for the mother’s health. The pregnant women in Non-newar were assisted by 45.45 percent mother in laws during delivery at home followed by Newar (44.44%). Similarly, most of Newar respondents (95.35%) used the safe delivery kit to cut the cord whereas only 77.19 percent Non-newar respondents used the safe delivery kit to cut the cord. Most of the women had suffered from health problem during delivery period like swelling of hands, face and legs; back paining and bleeding etc. Majority of the Newar mothers (97.67%) fed the first milk after the delivery followed by Non-newar (94.74%). Majority of respondents in both had used the foods containing ghee and meat during postnatal period. More than 85 percent Newar respondents took vitamin A capsule during postnatal period whereas Non-newar by only 82.46 percent. Similarly, most of the Newar respondents (48.84%) did not provide bath to their babies in fixed time followed by Non-newar respondents (45.61%). According to this study, 83.72 percent of Newar respondents and 78.95 percent of Non-newar respondents used family planning devices. 63.16 percent children had been suffered from different kinds of diseases in Non-newar community but only 58.14 percent children had been suffered from different kinds of diseases in Newar community. The contribution of husband and family members in antenatal, natal and postnatal period needs to be more effective which helps to reduce the maternal and child mortality. Therefore, there should have advocacy, awareness campaign and income based programs for the Jhaukhel community.