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   Table of Contents - Current issue
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July-September 2021
Volume 12 | Issue 3
Page Nos. 215-309

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REVIEW ARTICLE  

Efficacy and safety of potential vaccine candidates against coronavirus disease 2019: A systematic review p. 215
Subodh Kumar, Manoj Kumar Saurabh, Vikas Maharshi
DOI:10.4103/japtr.JAPTR_229_20  
Search for an effective and safe vaccine to prevent transmission of current pandemic is an unmet need. This study reviews and compares the available early phase clinical data of vaccine candidates which have reached phase 3 of clinical development. The latest update of “DRAFT landscape of coronavirus (CoV) disease 2019 candidate vaccines (October 2, 2020)” released by the World Health Organization was accessed to identify the potential vaccine candidates. The full text articles (published and/or preprint) of data of early clinical trials of the selected vaccines were accessed from the links provided in the same document, PubMed and/or medRxiv.com. After extraction and synthesis, the data were critically evaluated for the study efficacy and safety outcomes. Of the total 193 candidate vaccines 10 were found to reach phase 3 of the clinical development. Nine of these were included in the evaluation process. In all of the included studies, immunogenicity and serious adverse events/local or systemic adverse events/laboratory parameters abnormality was considered as efficacy and safety outcomes respectively. Immunogenicity response with most of the vaccines was either higher than or similar to the respective controls except one (recombinant adenovirus type 26 COV2 [Ad26.COV2.S]) for which it was less than that in control. Overall adverse events (related and/or unrelated) were more with vaccines than those with respective control(s) in three studies, in other two, these were similar whereas in one study, the events were less in the vaccine group than in control group and in the rest, data described were descriptive only without any mention for the same for the control. In conclusion all studies showed immunogenic response to target protein of severe acute respiratory syndrome CoV-2 and which was higher than the respective control except for Ad26.CoV2.S. Many of the vaccines caused more adverse events than the controls, however most were mild and transient and/or manageable.
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ORIGINAL ARTICLES Top

Anticancer, antioxidant, and antibacterial activities of the methanolic extract from Sphagneticola trilobata (L.) J. F pruski leaves Highly accessed article p. 222
Vivi Mardina, Syafruddin Ilyas, Halimatussakdiah Halimatussakdiah, Tisna Harmawan, Masitta Tanjung, Faridah Yusof
DOI:10.4103/japtr.JAPTR_131_21  
This study aims to investigate the potential of bioactive secondary metabolites contained in Sphagneticola trilobata (L.) J.F Pruski leaves as novel plant-derived anticancer agent. Qualitative bioactive compound contents in the methanolic extract of S. trilobata leaves were screened using phytochemical method. Antioxidant evaluation was carried out using 2,2-diphenyl-1-picrylhydrazyl assay; antibacterial – using well diffusion method on Escherichia coli and Salmonella typhi; and cytotoxicity – using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay on MCF-7 cell line and Vero Cell. It was found that the methanolic extract exhibited antioxidant activity with an IC50 value of 124.34 μg/mL. The inhibition zone values against E. coli and S. thypi (at extract concentration of 100 mg/mL) were 34.33 and 36 mm, respectively. In vitro MTT assay showed that our extract successfully reached 96% mortality with LC50 = 189.287 μg/mL, where the selective index of 2.5 suggest its selectivity against MCF-7 breast cancer cell line. In conclusion, the data of biological activities suggest the potential development of methanolic extract from S. trilobata leaves as a phytomedicine for breast cancer treatment.
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The effect of rice husk nanosilica hydroxyl compound on dentin biomineralization p. 227
Iffi Aprillia, Endang Suprastiwi, Aryo Megantoro, Luh Putu Trisna, Budi Utami, Sarmayana Yana
DOI:10.4103/japtr.JAPTR_227_21  
Rice husk nanosilica contains hydroxyl for dentin remineralization. The aim of this study was to analyze and correlate the ability of rice husk nanosilica to induce hydroxyapatite dentin. The detachment of hydroxyl from rice husk nanosilica was analyzed using the sol–gel and pyrolysis methods with Fourier transform infrared spectroscopy. Subsequently, exposing of the demineralized dentin to rice husk nanosilica was performed for a comparison. The formation of hydroxyapatite on dentin was analyzed using X-ray diffraction. The amount of hydroxyl released from the two methods was then correlated with the hydroxyapatite that formed at the dentin. The extraction of hydroxyl on rice husk nanosilica with two methods was the same. Analysis of the amount of hydroxyapatite dentin with both the methods corresponds to each other. The correlation test obtains the value of R = 0.656. Rice husk nanosilica has a similar capability to release hydroxyl compound and form hydroxyapatite dentin using two methods. The creation of hydroxyapatite dentin is not only caused by the exposure of rice husk nanosilica but also owing to other factors that might reinforce the process of hydroxyapatite formation.
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Antimutagenic activity of nanoparticles of Rhaphidophora pinnata leaves in mice using micronucleus assay p. 232
Masfria Masfria, Marianne Marianne, Yade Metri Permata, Steven Octavio, Sri Mulyani
DOI:10.4103/japtr.JAPTR_380_20  
Cancer is one of the deadliest diseases in the world. Cancer may occur due to gene mutation. Rhaphidophora pinnata is a plant that has many benefits, especially in the leaves which have been used traditionally to treat cancer. The aim of this research is to test the antimutagenic activity of nanoparticles R. pinnata using the micronucleus method. The mice were induced with cyclophosphamide and then followed with the administration of nanoparticles of R. pinnata at the doses of 50, 100, 200 mg/kg for 7 days. The antimutagenic activity was evaluated at the decrease in the number of micronucleus in 200 polychromatic erythrocytes (PCE) cells of mice bone marrow. The result showed that the reduction of amount of micronucleus in PCE of a negative control group, treatment groups, and normal group is 22.65%, 60.3%, 79.6%, 93.8%, and 100%. These results indicate that the antimutagenic activity of nanoparticle of R. pinnata increases proportionally as the doses were increased. It can be concluded that nanoparticles R. pinnata at the doses of 50, 100, and 200 mg/kg have antimutagenic activity.
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In silico estrogen receptor alpha antagonist studies and toxicity prediction of Melia azedarach leaves bioactive ethyl acetate fraction p. 236
Martha Ervina, Mohammad Rizki Fadhil Pratama, Hadi Poerwono, Juni Ekowati, Retno Widyowati, Katsuyoshi Matsunami, Sukardiman
DOI:10.4103/japtr.JAPTR_198_21  
The estrogen hormone dependent accounts for a major cause in the incidence of women breast cancer. Thus, their receptor, especially the estrogen receptor α (ER-α), is becoming a target in endocrine treatment. These ligand-inducible nuclear functions are regulated by an array of phytochemical and synthetic compounds, such as 17 β-estradiol and tamoxifen (4-hydroxytamoxifen [4OHT]). The Chinaberry (Melia azedarach) leaves are known naturally for relieving internal and external diseases. Previous studies revealed the potency of Melia's ethanolic extract and ethyl acetate fractions as anticancer; furthermore, this study aimed to resolve possible ER-α antagonist's mechanism and safety from M. azedarach leaves ethyl acetate fraction contents. Melia's phytochemical content was analyzed with electrospray ionization liquid chromatography-mass spectrometry, while its ER-α antagonist's potency was investigated by in silico. The computational docking was used to 3ERT (a human ER-α-4OHT binding domain complex) with Autodock Vina and related programs. The results presented Energy binding (ΔG) of Melia's quercetin 3-O-(2'',6''-digalloyl)-β-D-galactopyranoside was similar to 4OHT, and lower than its agonist 17 β-estradiol. Furthermore, the toxicity prediction of these compounds were revealed safer than 4OHT. The Melia's leaves ethyl acetate fraction, therefore, is a potential pharmacological material for further studies.
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Performance and drug deposition of kappa-carrageenan microspheres encapsulating ciprofloxacin HCl: Effect of polymer concentration p. 242
Dewi Melani Hariyadi, Tutiek Purwanti, Dinda Maulydia, Cindy Alicia Estherline, Esti Hendradi, Mahardian Rahmadi
DOI:10.4103/japtr.JAPTR_197_21  
It has been known that in respiratory disease, antibiotic is selected for respiratory diseases or lung infections and this research focused on ciprofloxacin HCl as a model. The aim was to evaluate the effect of kappa-carrageenan polymer concentrations on characteristics, release, and drug deposition in the lung. Ciprofloxacin HCl-carrageenan microspheres were produced with kappa carrageenan (0.75%, 0.50%, and 0.25%) as polymer and KCl (1.5%) as crosslinker. Physical characteristics were included morphology, size, moisture content, swelling index, mucoadhesivity, drug loading, entrapment efficiency, and yield. Freeze-dried microspheres were inhaled by animal, and drug deposition was observed. Results showed that dried, smooth, and spherical microspheres of size of 1.34 to 1.70 μm and loading of 15.63% to 38.72%. Entrapment efficiency and yield were 25.38%–51.61% and 52.53%–63.19%, respectively. Mucoadhesivity was 0.0059–0.0096 kg force, and release in 24 h was 74.38%–81.02%. Release kinetics demonstrated Higuchi mechanism. Increasing carrageenan concentration affected size, loading, and efficiency but did not influence adhesivity, yield, and release. Higher amount of polymer caused the lower deposit on the lungs. Respirable size of ciprofloxacin HCl-kappa carrageenan microspheres was successfully achieved target site and prolonged residence time in lungs.
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Encapsulation mechanism of α-mangostin by β-cyclodextrin: Methods of molecular docking and molecular dynamics p. 250
Dian Triwahyuningtyas, Sandra Megantara, Tai Tze Hong, Muhammad Yusuf, Muchtaridi Muchtaridi
DOI:10.4103/japtr.JAPTR_298_20  
The study aimed to investigate the interaction of host-guest between α-mangostin and β-cyclodextrin (βCD) and also to calculate the energy of the complex system between α-mangostin with βCD for drug delivery using methods of 15 molecular dynamics and molecular docking. Simulation of molecular docking and molecular dynamics was utilized to determine molecular interactions and the complex system's bond energy. The docking simulation results showed that α-mangostin-βCD complex has a Gibbs energy value (ΔG) of −6.69 kcal/mol. The Gibbs energy value (ΔG) of molecular dynamics simulation from MMGBSA calculation showed the binding energy of α-mangostin-βCD – 11.73 kcal/mol.
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Phytochemical screening, antimalarial activities, and genetic relationship of 16 indigenous Thai Asteraceae medicinal plants: A combinatorial approach using phylogeny and ethnobotanical bioprospecting in antimalarial drug discovery p. 254
Desy Liana, Kanchana Rungsihirunrat
DOI:10.4103/japtr.JAPTR_238_21  
Emergence of artemisinin resistance leads the people to discover the new candidate for antimalarial drug. Combinatorial phylogeny and ethnobotanical approach may be useful to minimize the expenditure and time in laboratory testing. Seven hundred and thirty-three ethnomedicinal plants were listed from literature search. Obtained 340 internal transcribed spacer (ITS) sequences of plant list which met criteria were retrieved from GenBank NCBI and analyzed by MUSCLE and maximum likelihood phylogenetic test to generate the phylogenetic tree. Interactive phylogenetic tree was generated by Interactive Tree of Life (ITOL, https://itol.embl.de) and showed strong clustered pattern on Asteraceae. Afterward, 16 species of Asteraceae were selected to investigate the antimalarial activity, phytochemical, and genetic diversity. The presence of phytochemical was determined by standard method. DNA fluorescence-based assay was performed to determine the antimalarial activity against 3D7 Plasmodium falciparum. IC50 μg/mL was used to categorize antimalarial activity. On the other hand, ITS universal primer was used to amplify and sequence the obtained extracted DNA of tested plant by cetyltrimethylammonium bromide method. Phylogenetic analyses were performed by MAFFT and RAxML with automatic bootstrapping. ITOL and Adobe Illustrator were used to generate interactive phylogenetic tree. All species tested showed the presence of phenolics and flavonoids, whereas alkaloids and terpenoids were shown vary among tested extracts. Among 16 species tested, 1 species exhibited good-moderate (Sphaeranthus indicus, IC50 6.59 μg/mL), 4 weak (Artemisia chinensis, Artemisia vulgaris, Tridax procumbens, and Blumea balsamifera), and 3 very weak (Eupatorium capillifolium, Wedelia trilobata, and Vernonia cinerea). Generated phylogenetic tree by ITS data was able to separate the tested species into their tribal classification. In addition, new medicinal properties of A. chinensis were discovered. Combining phylogeny approach with ethnobotanical data is useful to narrow down the selection of antimalarial plants candidate.
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In silico identification of natural products from Centella asiatica as severe acute respiratory syndromecoronavirus 2 main protease inhibitor p. 261
Putu Gita Maya, Widyaswari Mahayasih, Harizal , Herman , Islamudin Ahmad
DOI:10.4103/japtr.JAPTR_297_20  
Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) main protease (S-CoV-2 Mpro) is one of the main targets in designing antiviral against SARS-CoV-2. Centella asiatica contains several triterpenoids, polyacetylenes, and benzoic ester derivative with various biological activities including anti-inflammation and antiviral. Triterpenoids from C. asiatica could act as inhibitors of S-CoV-2 Mpro. The main objective of this study was to identify potential natural products from C. asiatica as S-CoV-2 Mpro inhibitor with better pharmacokinetic through in silico molecular docking method. : As much as 11 compounds from C. asiatica were docked with S-CoV-2 Mpro (PDB ID: 6LU7) using AutoDock v4.2.6. Pharmacokinetic parameters of these compounds were assessed using SwissADME (free access webserver). Molecular docking results of 11 natural products indicated that asiatate 6 and asiatate 10 have strong interaction with quite similar binding free energy compared to native ligand (‒9.00 and‒9.58 kcal/mol compared to ‒9.18 kcal/mol, respectively) with proper interaction to the catalytic dyad (His41 and Cys145). Pharmacokinetic analysis revealed that asiatate 4, asiatate 10, and asiatate 11 have poor pharmacokinetic properties. These results indicated that asiatate 6 could be recommended for further study as S-CoV-2 Mpro inhibitor.
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Development and validation of an automated solid-phase extraction-LC-MS/MS method for the bioanalysis of fluoxetine in human plasma p. 267
Ishtiaque Ahmad, Zabih Ullah, Mohd Ibrahim Khan, Abdullah Khaloofa Alahmar, Mohd Faiyaz Khan
DOI:10.4103/japtr.JAPTR_308_20  
A wide-range, specific, and precise liquid chromatography tandem mass spectrometric (LC-MS/MS) technique for quantifying fluoxetine (FLX) in human plasma was developed using the RapidTrace® automated solid-phase extraction (SPE) method; the analyte and internal standard (IS) were extricated on Oasis MCX SPE cartridges. Acetonitrile and 5 mM ammonium formate buffer (90:10 v/v) were used as mobile phase to achieve chromatographic separation on the reverse phase (C<sub>18</sub> column). The analyte and IS were ionized using +ve electrospray ionization approach which was further traced by multiple-reaction monitoring on a tandem mass spectrometer. To quantify the FLX and FLX-d5, the parent-to-daughter ion transition of m/z of 310.0/44.1 and 315.0/44.0 was used, respectively. The method demonstrated a linear active limit of 0.20–30 ng/ml with recoveries ranging from 63.04% to 79.39% for quality control samples and 61.25% for IS samples. The concentrations over the calibration range demonstrated acceptable precision and accuracy. Due to the high inconsistency of the FLX concentration data, the minimum threshold of the assay was kept at 0.20 ng/ml. The flow rate was maintained at 500 <Symbol>μ</Symbol>L/min, and the time for sample analysis for each injection was 3.5 min. The method was found to be specific, sensitive, and faster with minimum utilization of organic solvents and was utilized further for metabolic and pharmacokinetic studies.
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The color improvement of postdebonding white spot lesions after fluoride and casein phosphopeptide–amorphous calcium phosphate application p. 274
Haru Setyo Anggani, Putri Arifiani, Erwin Siregar
DOI:10.4103/japtr.JAPTR_203_21  
White spots are common side effects of orthodontic treatment, and their presence after debonding appears unesthetic. This study aimed to quantify and compare the visual improvement in postdebonding white spot lesions following fluoride and casein phosphopeptide–amorphous calcium phosphate (CPP–ACP) application. The sample included 42 upper premolars extracted for orthodontic reasons. Universal premolar brackets were bonded to the facial surfaces of the teeth that were exposed to a demineralization solution to create artificial white spot lesions, after which the brackets were debonded. The specimens were randomly allocated to three treatment groups (n = 14, each): acidulated phosphate fluoride (APF) gel application group; CPP–ACP paste application group; and control group. pH cycling was conducted to all groups for 14 days. Quantitative measurements were carried out using a spectrophotometer at the following times: before and after the white spot lesions artificially formed and after treating them. All groups showed significant differences in color change before and after treatment. However, there was no significant difference in the color improvement between the APF gel and CPP–ACP paste application groups. Although CPP–ACP application improved the color of the white spot lesions, it did not differ significantly from that of fluoride application.
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3,4-dimethoxychalcone novel ultraviolet-A-protection factor in conventional sunscreen cream p. 279
Erlina Fatmasari, Abdul Karim Zulkarnain, Rina Kuswahyuning
DOI:10.4103/japtr.JAPTR_89_21  
UltravioletA (UVA) rays with an intensity of 95% can induce skin cancer due to the activation of reactive oxygen species (ROS). The 3,4-dimethoxychalcone (3,4-DMC) chalcone derivative has a wide wavelength, antioxidant activity, presumed has activity as sunscreen (UVA rays). Topical delivery of water-insoluble 3,4-DMC with log P 3.84 required capable, cream formulation was chosen because it was suitable for application this chemical sunscreen. This study aims to obtain the optimal formulation of 3,4-DMC in a sunscreen cream dosage form as a UVA-protection factor (UVA-PF). This study involves experimental design. The cream 3,4-DMC was evaluated physically for 4 weeks by measuring pH, viscosity, spreadability, adhesion, centrifugation, freeze–thaw, photostability, UVA-PF used TranporeTM tape, and skin irritation test on animals. The result obtained was evaluated statistically using ANOVA (SPSS version 24). The ratio UVA/UVB value of 3,4 DMC sunscreen cream having 5 stars (*****) for all concentrations, shows the product in this study can be used as an anti-UVA agent in sunscreen cream cosmetic products. The stability of the cream has pH 4.0–4.2; spreadability 5–6 cm; viscosity 4.470–5.763; and adhesion <1 s. Freeze-thaw and centrifugation were known did not affect the stability due to the absence of separation. There was no wavelength shift in the photostability test and no skin irritation due to in vivo examination using New Zealand rabbits. The 3,4-DMC as a new agent in conventional sunscreen cream dosage form has good properties as a protection against UVA rays.
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The interaction of alpha-mangostin and its derivatives against main protease enzyme in COVID-19 using in silico methods p. 285
Syahrul Hidayat, Faisal Maulana Ibrahim, Kelvin Fernando Pratama, Muchtaridi Muchtaridi
DOI:10.4103/japtr.JAPTR_299_20  
More than 111 million people worldwide have been affected by the COVID-19 outbreak caused by SARS-CoV-2. The main therapeutic target of COVID-19 is main protease (Mpro). It plays a key role as an enzyme in the SARS-CoV-2 replication and transcription. In this case, the alpha-mangostin potentially has antiviral activity against Mpro by inhibiting this enzyme. Nevertheless, the alpha-mangostin has low solubility and a lack of information about alpha-mangostin activity against the SARS-CoV-2. The aim of this study is to describe the molecular interactions and identify the pharmacokinetics profile between alpha-mangostin and its derivatives. in silico study was conducted by pharmacokinetics and toxicity prediction, molecular docking simulation, and Lipinski's rule of five. FKS9 has a Gibbs free energy value of-10.5 kcal/mol with an inhibition constant of 36.45 μM and an interaction with amino acid His41 residue. Its human intestinal absorption and Caco-2 values were 95.13% and 47.71% while the plasma protein binding and blood-brain barrier values were 96.66% and 6.99%. FKS9 also has no mutagenic and carcinogenic potential. FKS9 as an alpha-mangostin derivative had the best interaction with the Mpro enzyme and its pharmacokinetic profiles was identified.
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Standardization of Leonurus sibiricus L. aerial part and capillary electrophoresis quantitative analysis of its leonurine content p. 291
Maneewan Suwatronnakorn, Somchai Issaravanich, Chanida Palanuvej, Nijsiri Ruangrungsi
DOI:10.4103/japtr.JAPTR_243_21  
The quality parameters of Leonurus sibiricus L. aerial part crude drugs were evaluated. Fifteen crude drugs were collected from various locations throughout Thailand. The transverse section of the stem of L. sibiricus showed quadrangular character highlighted the ribs with angular collenchyma. The epidermis was uniseriate with abundant glandular trichomes distribution. Prismatic calcium oxalate prisms were found in the stem medullary parenchyma. The histological character of crude drug powder showed bordered pitted vessel, fragment of fiber, glandular trichome, prism crystal, spiral vessel, starch granule, and stomata. The loss on drying, total ash, acid-insoluble ash, and moisture contents should be not more than 8.18, 15.28, 4.04, and 8.91 g/100 g dry weight, whereas ethanol and water-soluble extractive values should be not less than 7.67, and 17.21 g/100 g of dry weight, respectively. Leonurine in the crude drugs were analyzed by capillary electrophoresis (CE) with photodiode array detector. The ethanolic extraction performed by Soxhlet apparatus yielded 18.86 ± 4.09 g/100 g dry weight. The electropherogram detected at 277 nm showed the migration time of leonurine at 6.2 min. The developed CE was found to be valid for leonurine quantification in L. sibiricus ethanolic extract. The contents of leonurine in 15 crude drugs ranged from 0.79 to 4.23 mg/g with the average of 2.38 ± 1.10 mg/g dry weight. This study established the pharmacognostic specification of L. sibiricus crude drug in Thailand with special reference to a bioactive compound, leonurine. CE was beneficial technique for the analysis of leonurine in L. sibiricus aerial parts.
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Working goal of Brazilein sappan wood as a candidate for SARS-coV-2 antivirus drug against spike (S) glycoprotein, papain-like proteinase, and main protease: In silico study p. 298
Entuy Kurniawan, Dwi Krihariyani
DOI:10.4103/japtr.JAPTR_222_21  
Brazilein sappan wood, played by Spike (S) glycoprotein, Papain-Like proteinase (PLpro), and Main protease (Mpro), is expected to be a candidate for the antiviral drug SARS-CoV-2, which can inhibit viral attachment to the human body, replication, and transcription processes. The aim of this study was to predict in silico, using the comparative drug hydroxychloroquine, the working goal of brazilein sappan wood as a candidate for the antiviral drug SARS-CoV-2 against protein S, PLpro, and Mpro. The approach used is the in silico docking test using the computer program Molegro Virtual Docker. Receptor used by protein S, Protein Data Bank (PDB) code: 6M0J, NAG_601[E] ligand; PLpro, PDB code: 7JIT, Y95_501[A] ligand; and Mpro, PDB code: 1WOF, I12_1145[A] ligand. Data analysis was carried out by comparing the docking bond energies between the ligands at the target receptor. Silico test results for protein S: ligand bond energy NAG_601 [E] = −59.4555, brazilein = −71.5537, hydroxychloroquine = −79.3704; PLpro protein: Ligand bond energy Y95_501 [A] = −129.561, brazilein = −94.9761, hydroxychloroquine = −100.984; Mpro protein: Ligand bond energy I12 1145 [A] = −141.135, brazilein = −96.6169, hydroxychloroquine = −104.88. The above test results indicate that brazilein sappan wood has potential as a SARS-CoV-2 drug candidate, has a stable bond, and that the biological activity of the compound is stronger against S protein than the proteins of PLpro and Mpro.
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Characterization and acute oral toxicity of concentrated minerals of Pamekasan Madura seawater p. 305
Syafika Alaydrus, Abd Kakhar Umar, Sriwidodo Sriwidodo, Ajeng Diantini, Nasrul Wathoni, Riezki Amalia
DOI:10.4103/japtr.JAPTR_250_20  
Indonesia is a maritime country with abundant seawater mineral content. One of the regions with the highest salt production is Pamekasan Madura. Minerals are known to have many roles and benefits for our bodies, such as regulating fluid balance and metabolism. Therefore, this study aimed to characterize the physicochemical and microbial properties of concentrated minerals obtained through solvent evaporation and salt deposition for ± 60 days. Acute oral toxicity examination was performed as a first step in determining the safety of concentrated minerals to be used as a raw material for drugs. Based on the test results, the concentrated mineral has a clear yellow color, salty taste, and a bit bitter, odorless, with a pH of 6.6 ± 0.21. Concentrated minerals have high mineral content with levels of potassium, sodium, magnesium, boron, and calcium being 44734.1598 ± 12950.4633, 33192.1198 ± 2699.3419, 8738.1388 ± 100.4894, 2092.5715 ± 60.3224, and 276.9704 ± 13.1133 mg/Kg, respectively. The results of microbiological analysis of untreated concentrated minerals (without antimicrobials or sterilization) showed that the total plate count was within limits, including coliform and Salmonella. However, the total mold and yeast levels exceed the threshold. Based on the results of acute oral toxicity testing, the concentrated mineral is practically nontoxic. With high mineral content and low toxicity, it can be concluded that the concentrated minerals from Pamekasan Madura seawater is potential to be used as a raw medicinal material.
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