HTTP 200 OK
Allow: GET, HEAD, OPTIONS
Content-Type: application/json
Vary: Accept
{
"count": 319,
"next": "https://jabet.bsmiab.org/articles/?format=api&page=27",
"previous": "https://jabet.bsmiab.org/articles/?format=api&page=25",
"results": [
{
"id": 202,
"slug": "178-1589274220-prediction-of-immune-pressure-on-hiv-1-regulatory-gene-tat-by-human-host-through-bioinformatics-tools",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1589274220",
"recieved": "2020-04-23",
"revised": null,
"accepted": "2020-06-14",
"published": "2020-06-27",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/55/178-1589274220.pdf",
"title": "Prediction of immune pressure on HIV-1 regulatory gene tat by human host through bioinformatics tools",
"abstract": "<p>In the early establishment of HIV-1 infection, Tat protein plays an essential role in controlling other genes of HIV-1 (e.g., vif, vpr, vpu, nef, gag, pol and env), for viral pathogenesis, while maintaining its polymorphic nature. It is well documented that polymorphism of HIV-1 genome are created to escape immune pressures (e.g. CD8+, CD4+, B, NK-cells and others) by human host, during the course of infection. Over the time those mutations are incorporated or left in the HIV genome as an escaped flag or signature amino acid, and this scenario could be predictable using contemporary bioinformatics tools. Our sequence analysis from global database (LANL) revealed that Tat protein under positive immune pressure as dn/ds >1.5, even though differential immune pressure exists among the HIV-1 subtype. Average entropy score is 0.31, implying the less variable nature of this protein while amino acid variations are higher in C-terminal. Remarkably, the region encompassing by positions 38 to 51 amino acids are relatively conserved, over the year 2009 to 2017 across HIV-1 subtype. Indeed, subtype-specific SNP or signature amino acids were observed in various position of Tat, dominantly in C-terminal end. Our epitope density plot analysis, highlighted CTLs/CD8+ cells play a major role on Tat sequence variation around the globe. Taken together, our analysis illustrated the dynamic nature of polymorphism within HIV-1 proteins, which could be predictable to see the immune-mediated selective pressure by human host on viral genome.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 233-240.",
"academic_editor": "Dr. Md. Abdul Hannan, Dongguk University, South Korea.",
"cite_info": "Hasan Z, Hasan M, et al. Prediction of immune pressure on HIV-1 regulatory gene tat by human host through bioinformatics tools. J Adv Biotechnol Exp Ther. 2020; 3(3): 233-240.",
"keywords": [
"Bioinformatics",
"Entropy",
"Dn/ds",
"HIV-1 tat",
"Immune pressure"
],
"DOI": "10.5455/jabet.2020.d129",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>One of the notorious characteristics of HIV-1 virus is tremendous sequence variation, creating highly divergent circulating strains around the globe [<a href=\"#r-1\">1</a>]. Designing an ideal/universal vaccine against HIV-1 has been threatening to the scientific community due to this genetic variability and ability to escape a myriad of host immune responses [<a href=\"#r-2\">2</a>]. Remarkably, HIV-1 creates several mutations in its genome to escape immune-mediated selective pressure (e.g. CD8+, CD4+, NK, B-cells, host factor and others) by the human hosts during the course of infection [<a href=\"#r-3\">3-6</a>]. Although, over the time those mutations are incorporated or left in the HIV sequence as a flag or signature amino acid, when HIV transmit to other human hosts while facing different immune profile and this scenario could be predictable using contemporary bioinformatics tools [<a href=\"#r-3\">3</a>]. To escape or adapt immune pressure, HIV-1 acquire mutation (i.e. synonymous and nonsynonymous) and try to escape host responses while maintaining their fitness effects as minimal as possible.<br />\r\nIn the early establishment of HIV-1 infection Tat protein plays an important controlling role of other HIV-1 gene expressions (e.g. <em>vif, vpr, vpu, nef, gag, pol</em> and <em>env</em> ) for viral replication and disease pathogenesis [<a href=\"https://www.bsmiab.org/jabet/178-1589274220-prediction-immune-pressure-hiv-1-regulatory-gene-tat-human-host-bioinformatics-tools/#_ENREF_7\">7</a><a href=\"#r-7\">, 8</a>]. Despite its important role, it is evident that Tat is also variable proteins in HIV proteome [<a href=\"#r-9\">9</a>], indicating Tat is under immune pressure and evolving to adapt in the respective host system to act as a functional protein.<br />\r\nSeveral studies have shown that sequence polymorphism of HIV-1 Tat exists among the subtype (B, C, E and BF) of HIV-1 [<a href=\"#r-10\">10</a>]. Interestingly subtype C Tat exhibits greater transcriptional activity in the CD4<sup>+ </sup>T cell line compared with subtypes B and E and this is because of variations in positions 57 (Ser to Arg) and 63 (Glu to Gln), suggesting the Tat sequence variability has a significant advantage on HIV-1 subtype C replication [<a href=\"https://www.bsmiab.org/jabet/178-1589274220-prediction-immune-pressure-hiv-1-regulatory-gene-tat-human-host-bioinformatics-tools/#_ENREF_10\">10</a><a href=\"#r-10\">, 11</a>]. Therefore, the adaptation of sequence polymorphism among various HIV-1 subtypes in different subcontinents, highlighting host immune profile may be one of the driving forces of these polymorphisms, which, however, is one of the setbacks of designing effective vaccine so far. Thus, a comprehensive analysis of the dynamics of polymorphisms in HIV-1 proteins is a powerful tool to reveal actual interactions between HIV-1 and the host immune system. Moreover, to decipher HIV-1 <em>tat</em> gene sequence evolution and possible candidate target in vaccination strategy, it is inevitable to see how this gene being imposed with selective pressure by the human host, which could be predicted using publicly open-access portal of HIV sequence and immunology database along with the contemporary bioinformatics tools from Los Alamos National Laboratory (LANL) (<a href=\"https://www.hiv.lanl.gov/\">https://www.hiv.lanl.gov</a>).</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Sequence database</strong><br />\r\nAll the deposited <em>tat</em> gene nucleotide (intact ORF) sequence from the year 2009 to 2017 was retrieved from the Los Alamos National Laboratory (LANL) HIV sequence database. The HIV database contain comprehensive data on HIV genome sequences and allow to access large number of bioinformatical tools that can be used to analyze and visualize the dataset. In brief, from this website (https://www.hiv.lanl.gov), anyone can choose from the options button to define:- [i] Alignment type (web, filtered, subtype, compendium, and consensus/ancestral), [ii] Organism (HIV-1/2 or SIV), [iii] Region of the genome (env, nef, vif, vpr, cpu, and others), [iv] Subtype (A-K and recombinants), [v] DNA/Protein, [vi] Year, [vii] Format (FASTA, Clustal, Phylip, MEGA and others ) and get alignment file on their gene of interest.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Sequence alignment</strong><br />\r\nRetrieved sequence dataset was edited in MEGA7 (Molecular Evolutionary Genetics Analysis) software, an alignment tool for sequence manipulation [<a href=\"#r-12\">12</a>]. Sequences were spilt-up (n=150 to 200/subtype, each country not more than 10 sequences) based on subtype variation (A, B, C, D, F, G, H CRFs (AE, BC and AG) and CPX. Gaps were removed from the above selected multiple sequences in notepad. Finally, the non-align multiple sequence datasets were aligned again with Clustal W in MEGA7 [<a href=\"#r-13\">13</a>] with respect to consensus reference subtype sequence (latest year 2004) from LANL (<a href=\"https://www.hiv.lanl.gov/content/sequence/NEWALIGN/align.html#consensus\">https://www.hiv.lanl.gov/</a>).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Selective pressure and dn/ds ratio</strong><br />\r\nTo find the natural selection pressure on <em>tat</em> gene, dn/ds ratio was used using SNAP v2.1.1 [(synonymous (ds) vs non-synonymous (dn)] tools from LANL, which calculates dn and ds substitution rates based on a set of codon-aligned nucleotide sequences [<a href=\"#r-1\">1</a>]. In briefly, multiple aligned nucleotide sequence as FASTA files were uploaded in synonymous non-synonymous analysis site (<a href=\"https://www.hiv.lanl.gov/\">https://www.hiv.lanl.gov/</a>) to get the dn and ds values for each 101 position. Subsequently the average value of dn was divided with average ds to get dn/ds ratio. If dn/ds ratio ≥ 1, Positive selection and pathogen try to escape from host immune surveillance while dn/ds ratio < 1, Negative/Purifying selection and pathogen try to adapt to host immune system.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Variability of amino acid sequence of Tat protein</strong><br />\r\nA Shannon entropy score for each position in the Tat protein was calculated to see the extent of amino acid sequence variability [<a href=\"#r-9\">9</a>]. In this application, multiple aligned amino acid sequence as FASTA files from Clustal W (translated to amino acid) were uploaded in the Shannon Entropy-One site to get the frequency of each position as column in a sequence alignment independently. As such, this entropy tool assigns a score to each column that reflects the variability in that column. Finally, all the values for each position of Tat protein (ORF, 110 aa) were processed in excel and prism file for further analysis.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Epitope density plots</strong><br />\r\nHIV molecular immunology database is a summary of all HIV-1 epitopes, which have been reported in the literature, including HIV-1 Cytotoxic (CTL/CD8+), T-helper/CD4+ and Antibody/B-cells epitopes sites. Density plot illustrated the number of reported human HIV-1 (<a href=\"https://www.hiv.lanl.gov/content/immunology/index\">https://www.hiv.lanl.gov/</a>) epitopes spanning each amino acid of the targeted protein. In brief, the number of reported epitopes to each amino acid (aa) position of Tat protein (1-101 aa), from this database was recovered that already been positioning with HIV-1 reference strain HXB2. All the numerical values for T cells (CD4+ and CD8+) and B-cells were transferred to excel file followed by GraphPad Prism to create the figure and analysis, if required.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Statistical analysis</strong><br />\r\nAll the graphs and figures were generated using multivariate statistical package GraphPad Prism software (Version 6.0, La Jolla, CA, United States) and Excel. A <em>p</em> value <0.05 indicated by asterisk as significant, otherwise <em>p</em> value >0.05, non-significant using student’s t-test (two-tailed).</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><strong>Host mediated selective pressure on HIV-1 <em>tat</em> gene</strong><br />\r\nThe overall immune pressure on HIV-1 tat gene across the subtype is positive, dn/ds ratio ≥ 1. The average dn/ds value is more than 1.5 over the year from 2009 to 2017 but varied among the subtypes (<a href=\"#figure1\">Figure 1</a>A and 1B). This result suggests that, <em>tat</em> gene of circulating HIV-1 strains around the globe undergoing immune pressure by the human host, because the rate of substitutions at non-silent sites (dn, experienced selection) are higher than the rate of substitutions at silent sites (ds, presumed neutral). In particular, our analysis suggests that <em>tat</em> may experience substantially stronger selective forces, hence selection pressure and dn/ds is qualitatively different for samples drawn from a single population compared to sampled from various or mixed population [<a href=\"#r-14\">14</a>]. Interestingly, only subtype G showed lower dn/ds ratio (mean=1.1) over the year, while other subtypes did not. In addition, the prevalent subtypes responsible for global HIV/AIDS are B (North America, Europe, and Australia) and C (South Africa and South East Asia) [<a href=\"#r-15\">15</a>], have shown statistically significant difference of dn/ds ratio (B, mean=1.71; C, mean=1.49) over the year 2009 to 2017 on <em>tat</em> gene (<a href=\"#figure1\">Figure 1C</a>). Taken together, our analysis suggests that the differential adaptation of HIV-1 <em>tat</em> gene across the globe, due to the variation of human immune profile and imposed pressure.</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"536\" src=\"/media/article_images/2024/11/14/178-1589274220-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1.</strong> dn/ds ratio among HIV-1 subtype from 2009 to 2017. Autologous sequences of HIV-1 subtype A, B, C, D, F, G, H, AE, AG, BC and Cpx were retrieved from Los Alamos National Laboratory (LANL). (A) dn/ds ratio was calculated among the clades of HIV-1, (B) average dn/ds ratio was compared among the years from 2009 to 2017, (C) comparison of dn/ds ratio between clade B and C over the year 2009 to 2017 which is statistically significant using student’s t-test (p< 0.05).</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Variability of Tat protein and signature amino acid</strong><br />\r\nNext, we analyzed the variability of amino acid along the intact open reading frame (ORF) of Tat protein using Shannon entropy score [<a href=\"#r-9\">9</a>], which assesses the diversity in the population in a cross-sectional sense. Amino acid variations are not evenly distributed, and the average entropy score reached 0.31 over the year (<a href=\"#figure2\">Figure 2A-B</a>), confirming that Tat protein has not as variable as other proteins such as Env and Vpu [<a href=\"#r-9\">9</a>, <a href=\"#r-16\">16</a>]. Less variability was observed in domain I-IV, while higher in domain V-VI, an indication of the functional importance among the domains. Remarkably, position 38 to 51 (domain III and a partial region of domain IV) show relatively conserved (shaded area on<a href=\"#figure2\"> Figure 2A</a>) and these regions of Tat are known to play a role for functional internalization into the host cells [<a href=\"#r-17\">17</a>]. Sequence alignment of various HIV-1 subtype with consensus sequence from LANL database (HIV-1, M group) also revealed that Tat proteins are relatively conserved among the subtypes but contain subtype-specific signature amino acids, for example at position 57, Serine (S) for type C, while Arginine (R) for type B. Interestingly, substantial variation was observed at position 29, over 2009-2017 years, but this position also has signature amino acid for different subtypes, Lysine (K) for type A, B, D, H, Cpx; Histidine (H) for type C; Arginine (R) for type F, BC; Methionine (M) for type G; and Isoleucine (I) for type AE and AG (<a href=\"#figure3\">Figure 3</a>). Certainly, distributions of signature amino acids were more prominent in the C-terminal part of Tat than N-terminal, which is good agreement with our entropy analysis in <a href=\"#figure2\">Figure 2</a>.</p>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"479\" src=\"/media/article_images/2024/11/14/178-1589274220-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2. </strong>Entropy variation of HIV-1 group M from 2009 to 2017. (A) Shannon entropy was used to measure the relative variation in different positions or regions of an aligned protein of Tat from 2009 to 2017. (B) comparison of entropy level over the years of HIV-1 group M Tat protein.</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Immune cells targeted epitopes across Tat protein</strong><br />\r\nTo see how human CTL/CD8+, T-helper/CD4+ and Antibody/B- cells shape HIV-1 Tat protein, we aim to search literature for reported epitopes from HIV molecular immunology database of LANL. The number of reported epitopes on Tat protein is higher by CD8+ rather than CD4+ and B-cells (<a href=\"#figure4\">Figure 4</a>), suggesting that variability of Tat driven by CD8+ cells across the globe regardless of their subtype. We have also noticed the highly variable C-terminal region has less reported epitope by CTLs/CD8+ cells than its relatively conserved N-terminal, but the scenarios are not the same for CD4+ and B-cells.</p>\r\n\r\n<div id=\"figure3\">\r\n<figure class=\"image\"><img alt=\"\" height=\"114\" src=\"/media/article_images/2024/11/14/178-1589274220-Figure3.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 3. </strong>Sequence alignment of Tat protein of HIV-1 subtypes. Consensus Tat protein sequences of different subtypes (A, B, C, D, F, G, H, AE, AG, BC and Cpx) were aligned with consensus HIV-1 M group from 2004 using LANL database.</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"figure4\">\r\n<figure class=\"image\"><img alt=\"\" height=\"218\" src=\"/media/article_images/2024/11/14/178-1589274220-Figure4.jpg\" width=\"400\" />\r\n<figcaption><strong>Figure 4. </strong>Epitope density plots across Tat protein of HIV-1. Density plot of reported epitopes from the literature on Tat protein of HIV-1. Data were adapted from HIV molecular immunology database of LANL</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>Human immunodeficiency virus type-1 (HIV-1) displays extraordinary genetic diversity, which has been a major setback in the development of vaccine and antiretroviral drugs. During host-pathogen interaction, the virus escapes immune pressures while creating a mutation (SNP) or shed certain regions of HIV-1 (i.e. glycosylation), because of its importance for functional and structural conservation to replicate [<a href=\"#r-3\">3</a>, <a href=\"#r-18\">18</a>, <a href=\"#r-19\">19</a>]. Improvements in DNA sequencing technologies, variety of statistical tools and the availability of large sequence datasets of HIV-infected individuals (LANL), allow us to employ population-based genetic association in academia using HIV database tools (https://www.hiv.lanl.gov/content/sequence/HIV/HIVTools.html). In addition, the undertaken entire bioinformatics steps in this current study are summarized in <a href=\"#figure5\">Figure 5</a>.<br />\r\nWe have provided evidence that Tat protein under positive immune pressure by calculating dn/ds ratio (>1.5) using a global sequence database. Although, imposed differential immune pressure exists among the HIV-1 subtype, such as dn/ds ratio for subtype B is 1.71 whereas C is 1.49 over the year 2009 to 2017 (<a href=\"#figure1\">Figure 1</a>A and 1C). The amino acid variations are prominent in the C-terminal of Tat protein (<a href=\"#figure2\">Figure 2A</a>), even though the average entropy is 0.31, suggesting this protein is less variable than other proteins of HIV-1, for example Env, Gag, Pol, and Nef [<a href=\"#r-1\">1</a>]. Remarkably, a region of position 38 to 51, known to play an important role of HIV-1 internalization in host cells, relatively conserved, could be an intriguing target for vaccination. Moreover, certain SNPs or signature amino acids were observed among the subtype in various positions of Tat, dominantly in the C-terminal end (<a href=\"#figure3\">Figure 3</a>). The host immune profile may be one of the confounding factors of these signature amino acids or SNPs [<a href=\"#r-3\">3</a>]. Epitope density plot analysis along with the Tat protein by T- and B-cells, indicate that CTLs/CD8+ cells play a major role in Tat sequence variation, which, however, needs further clarification using viral killing assay.<br />\r\nHence, the development of a vaccine against HIV-1 infection represents the only realistic way to control the global expansion of the HIV-1 pandemic, especially in the developing world. Therefore, this study aimed at finding a delicate point of HIV-1 by quantifying selection pressures, identification of genetic loci undergoing adaptation/single nucleotide polymorphism (SNP) for subtype-specific landscape, finding the conserved region of gene <em>in vivo </em>[<a href=\"#r-3\">3</a>, <a href=\"#r-19\">19</a>]. Indeed, t­­argeting the signature SNP means targeting a very weak point of HIV-1 and this information would be useful for vaccine design focusing on HIV-1 subtype variation in the context of global perspective. Tat protein is a key player in HIV-1 infection since the virus lacking Tat has no infectivity as documented by several studies [<a href=\"#r-20\">20-22</a>]. Interestingly, vaccination with Tat antigen in rodents and monkeys (systemic and mucosal administration) has also shown that it can activate specific humoral and cellular (including CTLs) immune responses [<a href=\"https://www.bsmiab.org/jabet/178-1589274220-prediction-immune-pressure-hiv-1-regulatory-gene-tat-human-host-bioinformatics-tools/#_ENREF_23\">23</a><a href=\"#r-23\">, 24</a>]. Therefore, finding SNPs and/or immunogenic conserved regions may be one of the contemporary approaches to target HIV-1, to boost the current vaccine efficacy.</p>\r\n\r\n<div id=\"figure5\">\r\n<figure class=\"image\"><img alt=\"\" height=\"350\" src=\"/media/article_images/2024/11/14/178-1589274220-Figure5.jpg\" width=\"334\" />\r\n<figcaption><strong>Figure 5. </strong>Flow diagram of entire steps. Graphical representation of complete processes which has been taken in this study are summarized.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 5,
"section_title": "CONCLUSIONS",
"body": "<p>The HIV sequence and immunology databases from Los Alamos National Laboratory is the repository of information about autologous sequences, alignments, epitopes and antibody binding sites along with associated tools for students and researchers studying HIV. Noteworthy, these datasets were generated and published by various laboratories around the world from HIV-1 infected human patients. Therefore, in this current study we have shown here, how to use these information’s from this open resource (e.g. LANL) to draw a prediction map/dynamics (Figure 5) of immune pressure on HIV-1, as a model pathogen.</p>"
},
{
"section_number": 6,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>We like to thank SUST Research Centre for their funding and other technical support all through this study. We also like to thank administrative and technical staffs in Dept. of Biochemistry and Molecular biology, SUST, to set up computer and Internet facilities to complete the sequence analysis. We like to special thanks to Tanvir Hossain, Lecturer, for his kind support in the computer lab and valuable suggestions, Dept. of Biochemistry and Molecular biology, SUST, Sylhet, Bangladesh.</p>"
},
{
"section_number": 7,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>ZH, MH and AA were involved in conception and design of the experiments. MH, AA, RM, MAN and ZSN contributed to perform the sequence analysis. ZH, MH and AA analyzed data. ZH wrote the manuscript and MH and AA help in revising the manuscript. MAH and MWM contributed to revising it critically for important intellectual content. ZH made the final approval of the version to be published.</p>"
},
{
"section_number": 8,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/11/14/178-1589274220-Figure1.jpg",
"caption": "Figure 1. dn/ds ratio among HIV-1 subtype from 2009 to 2017. Autologous sequences of HIV-1 subtype A, B, C, D, F, G, H, AE, AG, BC and Cpx were retrieved from Los Alamos National Laboratory (LANL). (A) dn/ds ratio was calculated among the clades of HIV-1, (B) average dn/ds ratio was compared among the years from 2009 to 2017, (C) comparison of dn/ds ratio between clade B and C over the year 2009 to 2017 which is statistically significant using student's t-test (p< 0.05).",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/11/14/178-1589274220-Figure2.jpg",
"caption": "Figure 2. Entropy variation of HIV-1 group M from 2009 to 2017. (A) Shannon entropy was used to measure the relative variation in different positions or regions of an aligned protein of Tat from 2009 to 2017. (B) comparison of entropy level over the years of HIV-1 group M Tat protein.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/11/14/178-1589274220-Figure3.jpg",
"caption": "Figure 3. Sequence alignment of Tat protein of HIV-1 subtypes. Consensus Tat protein sequences of different subtypes (A, B, C, D, F, G, H, AE, AG, BC and Cpx) were aligned with consensus HIV-1 M group from 2004 using LANL database.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/11/14/178-1589274220-Figure4.jpg",
"caption": "Figure 4. Epitope density plots across Tat protein of HIV-1. Density plot of reported epitopes from the literature on Tat protein of HIV-1. Data were adapted from HIV molecular immunology database of LANL",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/11/14/178-1589274220-Figure5.jpg",
"caption": "Figure 5. Flow diagram of entire steps. Graphical representation of complete processes which has been taken in this study are summarized.",
"featured": false
}
],
"authors": [
{
"id": 890,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Zafrul",
"family_name": "Hasan",
"email": "zafrul-bmb@sust.edu",
"author_order": 1,
"ORCID": null,
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Zafrul Hasan, Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh. E-mail: zafrul-bmb@sust.edu",
"article": 202
},
{
"id": 891,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Mahedi",
"family_name": "Hasan",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 202
},
{
"id": 892,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Arafat Islam",
"family_name": "Ashik",
"email": null,
"author_order": 3,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 202
},
{
"id": 893,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Rumana",
"family_name": "Mahtarin",
"email": null,
"author_order": 4,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 202
},
{
"id": 894,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Md. Ali",
"family_name": "Newaj",
"email": null,
"author_order": 5,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 202
},
{
"id": 895,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Zakia Sultana",
"family_name": "Nishat",
"email": null,
"author_order": 6,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 202
},
{
"id": 896,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Mohammad Abul",
"family_name": "Hasnat",
"email": null,
"author_order": 7,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 202
},
{
"id": 897,
"affiliation": [
{
"affiliation": "Dept. of Biochemistry and Molecular Biology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh."
}
],
"first_name": "Md. Waseque",
"family_name": "Mia",
"email": null,
"author_order": 8,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 202
}
],
"views": 631,
"downloads": 100,
"references": [
{
"id": 6806,
"serial_number": 1,
"pmc": null,
"reference": "Korber-Irrgang B. HIV signature and sequence variation analysis. computational analysis of HIV molecular sequences. Allen G Rodrigo and Gerald H Learn, eds Dordrecht, Netherlands: Kluwer Academic Publishers. 2000;Chapter 4:55-72.",
"DOI": null,
"article": 202
},
{
"id": 6807,
"serial_number": 2,
"pmc": null,
"reference": "Mascola JR, Haynes BF. HIV-1 neutralizing antibodies: understanding nature’s pathways. Immunological reviews. 2013;254:225-44.",
"DOI": null,
"article": 202
},
{
"id": 6808,
"serial_number": 3,
"pmc": null,
"reference": "Brumme ZL, John M, Carlson JM, Brumme CJ, Chan D, Brockman MA, et al. HLA-associated immune escape pathways in HIV-1 subtype B Gag, Pol and Nef proteins. PloS one. 2009;4:e6687.",
"DOI": null,
"article": 202
},
{
"id": 6809,
"serial_number": 4,
"pmc": null,
"reference": "Jost S, Altfeld M. Evasion from NK cell-mediated immune responses by HIV-1. Microbes and infection. 2012;14:904-15.",
"DOI": null,
"article": 202
},
{
"id": 6810,
"serial_number": 5,
"pmc": null,
"reference": "Mujib S, Liu J, Rahman A, Schwartz JA, Bonner P, Yue FY, et al. Comprehensive Cross-Clade Characterization of Antibody-Mediated Recognition, Complement-Mediated Lysis, and Cell-Mediated Cytotoxicity of HIV-1 Envelope-Specific Antibodies toward Eradication of the HIV-1 Reservoir. Journal of virology. 2017;91.",
"DOI": null,
"article": 202
},
{
"id": 6811,
"serial_number": 6,
"pmc": null,
"reference": "Zheng N, Fujiwara M, Ueno T, Oka S, Takiguchi M. Strong ability of Nef-specific CD4+ cytotoxic T cells to suppress human immunodeficiency virus type 1 (HIV-1) replication in HIV-1-infected CD4+ T cells and macrophages. Journal of virology. 2009;83:7668-77.",
"DOI": null,
"article": 202
},
{
"id": 6812,
"serial_number": 7,
"pmc": null,
"reference": "Cafaro A, Tripiciano A, Picconi O, Sgadari C, Moretti S, Butto S, et al. Anti-Tat Immunity in HIV-1 Infection: Effects of Naturally Occurring and Vaccine-Induced Antibodies Against Tat on the Course of the Disease. Vaccines. 2019;7.",
"DOI": null,
"article": 202
},
{
"id": 6813,
"serial_number": 8,
"pmc": null,
"reference": "Brady J, Kashanchi F. Tat gets the “green” light on transcription initiation. Retrovirology. 2005;2:69.",
"DOI": null,
"article": 202
},
{
"id": 6814,
"serial_number": 9,
"pmc": null,
"reference": "Yusim K, Kesmir C, Gaschen B, Addo MM, Altfeld M, Brunak S, et al. Clustering patterns of cytotoxic T-lymphocyte epitopes in human immunodeficiency virus type 1 (HIV-1) proteins reveal imprints of immune evasion on HIV-1 global variation. Journal of virology. 2002;76:8757-68.",
"DOI": null,
"article": 202
},
{
"id": 6815,
"serial_number": 10,
"pmc": null,
"reference": "Li L, Dahiya S, Kortagere S, Aiamkitsumrit B, Cunningham D, Pirrone V, et al. Impact of Tat Genetic Variation on HIV-1 Disease. Advances in virology. 2012;2012:123605.",
"DOI": null,
"article": 202
},
{
"id": 6816,
"serial_number": 11,
"pmc": null,
"reference": "Kurosu T, Mukai T, Komoto S, Ibrahim MS, Li YG, Kobayashi T, et al. Human immunodeficiency virus type 1 subtype C exhibits higher transactivation activity of Tat than subtypes B and E. Microbiology and immunology. 2002;46:787-99.",
"DOI": null,
"article": 202
},
{
"id": 6817,
"serial_number": 12,
"pmc": null,
"reference": "Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular biology and evolution. 2016;33:1870-4.",
"DOI": null,
"article": 202
},
{
"id": 6818,
"serial_number": 13,
"pmc": null,
"reference": "Wrabl JO, Grishin NV. Gaps in structurally similar proteins: towards improvement of multiple sequence alignment. Proteins. 2004;54:71-87.",
"DOI": null,
"article": 202
},
{
"id": 6819,
"serial_number": 14,
"pmc": null,
"reference": "Ronsard L, Lata S, Singh J, Ramachandran VG, Das S, Banerjea AC. Molecular and genetic characterization of natural HIV-1 Tat Exon-1 variants from North India and their functional implications. PloS one. 2014;9:e85452.",
"DOI": null,
"article": 202
},
{
"id": 6820,
"serial_number": 15,
"pmc": null,
"reference": "Hemelaar J, Gouws E, Ghys PD, Osmanov S. Global and regional distribution of HIV-1 genetic subtypes and recombinants in 2004. Aids. 2006;20:W13-23.",
"DOI": null,
"article": 202
},
{
"id": 6821,
"serial_number": 16,
"pmc": null,
"reference": "Hasan Z, Carlson JM, Gatanaga H, Le AQ, Brumme CJ, Oka S, et al. Minor contribution of HLA class I-associated selective pressure to the variability of HIV-1 accessory protein Vpu. Biochemical and biophysical research communications. 2012;421:291-5.",
"DOI": null,
"article": 202
},
{
"id": 6822,
"serial_number": 17,
"pmc": null,
"reference": "Schwarze SR, Hruska KA, Dowdy SF. Protein transduction: unrestricted delivery into all cells? Trends in cell biology. 2000;10:290-5.",
"DOI": null,
"article": 202
},
{
"id": 6823,
"serial_number": 18,
"pmc": null,
"reference": "Silver ZA, Antonopoulos A, Haslam SM, Dell A, Dickinson GM, Seaman MS, et al. Discovery of O-Linked Carbohydrate on HIV-1 Envelope and Its Role in Shielding against One Category of Broadly Neutralizing Antibodies. Cell reports. 2020;30:1862-9 e4.",
"DOI": null,
"article": 202
},
{
"id": 6824,
"serial_number": 19,
"pmc": null,
"reference": "Moore CB, John M, James IR, Christiansen FT, Witt CS, Mallal SA. Evidence of HIV-1 adaptation to HLA-restricted immune responses at a population level. Science. 2002;296:1439-43.",
"DOI": null,
"article": 202
},
{
"id": 6825,
"serial_number": 20,
"pmc": null,
"reference": "Zhou M, Deng L, Kashanchi F, Brady JN, Shatkin AJ, Kumar A. The Tat/TAR-dependent phosphorylation of RNA polymerase II C-terminal domain stimulates cotranscriptional capping of HIV-1 mRNA. Proceedings of the National Academy of Sciences of the United States of America. 2003;100:12666-71.",
"DOI": null,
"article": 202
},
{
"id": 6826,
"serial_number": 21,
"pmc": null,
"reference": "Meltzer B, Dabbagh D, Guo J, Kashanchi F, Tyagi M, Wu Y. Tat controls transcriptional persistence of unintegrated HIV genome in primary human macrophages. Virology. 2018;518:241-52.",
"DOI": null,
"article": 202
},
{
"id": 6827,
"serial_number": 22,
"pmc": null,
"reference": "Nicoli F, Gallerani E, Sforza F, Finessi V, Chachage M, Geldmacher C, et al. The HIV-1 Tat protein affects human CD4+ T-cell programing and activation, and favors the differentiation of naive CD4+ T cells. Aids. 2018;32:575-81.",
"DOI": null,
"article": 202
},
{
"id": 6828,
"serial_number": 23,
"pmc": null,
"reference": "Tikhonov I, Ruckwardt TJ, Hatfield GS, Pauza CD. Tat-neutralizing antibodies in vaccinated macaques. Journal of virology. 2003;77:3157-66.",
"DOI": null,
"article": 202
},
{
"id": 6829,
"serial_number": 24,
"pmc": null,
"reference": "Gavioli R, Cellini S, Castaldello A, Voltan R, Gallerani E, Gagliardoni F, et al. The Tat protein broadens T cell responses directed to the HIV-1 antigens Gag and Env: implications for the design of new vaccination strategies against AIDS. Vaccine. 2008;26:727-37.",
"DOI": null,
"article": 202
}
]
},
{
"id": 201,
"slug": "178-1590167174-micropropagation-of-commercially-important-orchid-dendrobium-palpebrae-lindl-through-in-vitro-developed-pseudobulb-culture",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1590167174",
"recieved": "2020-04-27",
"revised": null,
"accepted": "2020-06-14",
"published": "2020-06-26",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/03/178-1590167174.pdf",
"title": "Micropropagation of commercially important orchid Dendrobium palpebrae Lindl. through in vitro developed pseudobulb culture",
"abstract": "<p><strong> </strong><em>Dendrobium palpebrae </em>Lindl. an epiphytic fragrant indigenous orchid species of Bangladesh and distributed in Bangladesh, eastern Himalayas, Myanmar, Thailand, Laos and Vietnam at altitudes of 800 to 2500 meters. From <em>in vitro</em> growing plantlets, upper and lower part of pseudobulb segments were aseptically cultured on MS medium supplemented with auxins (IAA, IBA, NAA, Picloram) and cytokinins (BAP, Kinetin). <em>In vitro </em>grown pseudobulb segments both upper and lower part directly produced multiple shoot buds <em>via</em> organogenesis. Average highest number of MSBs (8.21 ± 0.44/segment in lower part; 6.43 ± 0.40/segment in upper part) produced on MS medium with 1.0 mg/l NAA + 2.0 mg/l BAP followed by (7.24 ± 0.41/segment in lower part; 5.96 ± 0.37/segment in upper part) MS medium supplemented with 1.0 mg/l Picloram + 2.0 mg/l BAP. Highest individual shoot bud increased length was recorded on agar solidified MS medium with 1.0 mg/l Picloram + 1.0 mg/l BAP (3.76 ± 0.14 cm) followed by liquid MS + 0.5 mg/l NAA + 1.0 mg/l BAP (3.11 ± 0.12 cm) medium. Elongation of shoot bud was better in agar solidified media than liquid condition and MS was found better than PM. Shoot bud derived seedlings were best responses as increase in length as well as the number of roots developed on agar solidified MS with 0.5 mg/l NAA (4.82 ± 0.22 cm/shoot bud and 2.75 ± 0.17 no/shoot bud) followed by MS with 0.5 mg/l IBA supplemented medium (4.49 ± 0.12 cm/shoot bud and 2.57 ± 0.08 no/shoot bud). The plantlets were successfully transferred to outside environment by successive phases of acclimatization.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 225-232.",
"academic_editor": "Dr. Akhi Moni, ABEx Bio-Research Center, Bangladesh.",
"cite_info": "Bhowmik TK, Rahman MM. Micropropagation of commercially important orchid Dendrobium palpebrae Lindl. through in vitro developed pseudobulb culture. J Adv Biotechnol Exp Ther. 2020; 3(3): 225-232.",
"keywords": [
"Micropropagation",
"Dendrobium palpebrae",
"MSBs",
"PGRs",
"SPSs"
],
"DOI": "10.5455/jabet.2020.d128",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>Orchids are flowering plants, commercially grown worldwide as cut flower and potted plants in floriculture trade. Orchidaceae is considered as the most advanced and largest among monocotyledons, including most multifariousness family of flowering plants, consisting of more than 35,000 species belonging to 700-800 genera [<a href=\"#r-1\">1</a>]. Due to their ornamental and medicinal importance they demand a very high price in the international market [<a href=\"#r-1\">1</a>]. They are well known for their strange shaped, longevity and beautiful looking flowers. They are cosmopolitan but primarily distributed in tropical areas. They are rare in arctic regions [<a href=\"#r-1\">1</a>]. Many indigenous valued orchids are found in Bangladesh; mainly in Chittagong Hill Tracts, Chittagong, Cox’s Bazar, greater Sylhet, Gazipur and Sundarbans mangrove forest [<a href=\"#r-2\">2</a>].<br />\r\n<em>Dendrobium</em> is one of the largest genera of orchidaceae and contains about 1184 species worldwide [<a href=\"#r-3\">3</a>] and has high horticultural value. They are widely distributed and cultivated in the world ranging from southern Asia to New Guinea and Australia [<a href=\"#r-4\">4</a>]. In Bangladesh, about 27 species of<em> Dendrobium </em>are distributed throughout the country. <em>Dendrobium palpebrae </em>Lindl. distributed in Bangladesh, eastern Himalayas, Myanmar, Thailand, Laos and Vietnam at altitudes of 800 to 2500 meters. This fragrant indigenous epiphytic species blooms from spring to late summer and for commercially important as cut flower [<a href=\"#r-2\">2</a>].<br />\r\nOrchids can be easily propagated through <em>in vitro</em> propagation or tissue culture technique by using different plant parts as explants such as seeds, shoot tip, flower bud segment, lateral bud, young inflorescence, inflorescence, node, root, leaf and pseudobulb [<a href=\"#r-5\">5</a>]. Through micropropagation technique we can produce large number of disease-free plantlets at a very low cost. Micropropagation technique is appropriate for multiplication rather than <em>in vivo </em>[<a href=\"#r-6\">6</a>]. There is a great scope for large scale production of <em>Dendrobium</em> orchid in Bangladesh to meet the demand of international market and to earn foreign currency through export [<a href=\"#r-7\">7</a>]. The current study was intending to develop a reliable, reproducible and efficient protocol for mass multiplication of commercially high demanding orchid <em>D. palpebrae</em>.</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Explant and culture conditions</strong><br />\r\n<em>In vitro </em>developed seedlings of <em>Dendrobium palpebrae </em>Lindl. were used for micropropagation. For agar solidified media, agar (Himedia, India) was heated till is dissolved and finally mixing with stock solution gently. Plant Growth Regulators (Merck, Germany) <em>viz.</em> 6-benzyl Amino Purine (BAP), Kinetin (Kn), Picloram (Pic), Napthelene Acetic Acid (NAA), Indole Acetic Acid (IAA) and Indole Butaric Acid (IBA) were freshly prepared. 100 ml of the media were dispensed into 250 ml culture bottles (Duran, Germany) and autoclaved (HYSC, Korea) at 121<sup>0</sup>C for 20 minutes at 15 lbs pressure. p<sup>H</sup> (Fisons, UK) of the medium was set at 5.8 using 0.1N NaOH or HCl prior to gelling with agar and the culture temperature was maintained at 25±2 <sup>0</sup>C. Humidity level was between 50-60% and light was maintained between 4000-5000 lux illumination from cool white fluorescent to set 14/10 h photoperiod [<a href=\"#r-8\">8</a>]. Cultures were sub-cultured regularly and observed once in a week.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Micropropagation of pseudobulb segments</strong><br />\r\n<em>In vitro</em> grown pseudobulbs were cut 0.5 to 1.0 cm size using sterilized surgical blade and forceps under the laminar air flow cabinet (HYSC, Korea). Then the cuttings of upper and lower part were put into the culture vessel containing 0.8% (w/v) agar solidified MS based micropropagation media supplemented with sixteen different concentrations and combinations of PGRs.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Elongation of multiple shoot buds (MSBs)</strong><br />\r\nEighteen types of elongation media were prepared using full strength [<a href=\"#r-9\">9-10</a>] based solid & liquid media supplemented with different concentrations and combinations of PGRS. Full strength MS and based solid & liquid eighteen types of elongation media were prepared using with different concentrations and combinations of PGRs. 0.8% (w/v) agar was also used in solid media but in liquid media no agar was added.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Rooting of multiple shoot buds derived seedlings</strong><br />\r\nFor <em>in vitro</em> rooting of <em>D.</em> palpebrae, 0.8% (w/v) agar solidified half strength MS0 (Hormone free Murashige and Skoog medium) with 1.5% (w/v) sucrose and nine different types of MS medium supplemented with 3% (w/v) sucrose with three different concentrations of auxins <em>viz</em>. IAA, IBA and NAA were used for induction of strong and stout root system.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Hardening and transplantation</strong><br />\r\nFor hardening 90 days old plantlets with good rooting and 3-4 leaf conditions were selected. A gradual system of hardening was taken place in order to grow healthy plantlets. In this process, cultured vessels were kept open in the culture room for several hours, and then it was exposed to natural light for a day. Further, plantlets were washed by double distilled water to remove the adhering agar. Plants were treated with auxins to induce <em>ex vitro</em> rooting and roots were treated with fungicide. Then the seedlings of <em>D.</em> <em>palpebrae</em> were transferred to plastic pots containing a potting mixture of sterilized small brick, coal pieces, saw dust and peat moss at a ratio of 1: 1: 1: 0.5 and kept in the green house (at 25-30<sup>0</sup>C and RH 60-70%). Transplanted seedlings were watered regularly for about 2-3 months where the seedlings established and grew well.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Computation and presentation of data</strong><br />\r\n<em>In vitro </em>culture experiments were carried out systematically with the use of sufficient no of explant. Different growth parameters were considered to record data on morphogenic responses of explant under different conditions. The data on different parameters from different experiments were recorded after required days of culture.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Statistical analysis</strong><br />\r\nAll experiments were conducted in triplicate and data were presented as means ± standard error (mean ± SE). Standard deviations (SD) was calculated with Microsoft Excel 2013. In the table the mean ( ) data of different replication of each treatment are accompanied by standard error of mean (SE) which was calculated as follows:<br />\r\nStandard error (SE) = SD/root of N<br />\r\nWhere, SD = Standard deviation, N =Number of observations.</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><strong><em>In vitro </em>micropropagation of pseudobulb segments</strong><br />\r\nPseudobulbs were collected from <em>in vitro </em>derived seedlings as a source of explant of <em>Dendrobium palpebrae</em> for rapid micropropagation [<a href=\"#r-11\">11-13</a>] and result is shown on <a href=\"#Table-1\">Table 1</a>. Two types of pseudobulb segments; the upper part and the lower part were cultured on 0.8% (w/v) agar solidified MS media supplemented with various combinations and concentrations of PGRs and produced multiple shoot buds (MBSs) <em>via </em>direct organogenesis. The efficiency of a medium was assessed on the basis of number of shoot buds produced from each explant. Average highest number of MSBs (8.21 ± 0.44/segment in lower part; 6.43 ± 0.40/segment in upper part) produced on MS medium with 1.0 mg/l NAA + 2.0 mg/l BAP (<a href=\"#figure1\">Figure 1a</a>) followed by (7.24 ± 0.41/segment in lower part; 5.96 ± 0.37/segment in upper part) MS medium supplemented with 1.0 mg/l Picloram + 2.0 mg/l BAP. It’s also noted that lower part of pseudobulb segments showed better response than upper part.</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"758\" src=\"/media/article_images/2024/37/14/178-1590167174-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1. </strong>(1a-1f): <em>In vitro</em> micropropagation, shoot bud elongation, rooting, SPSs development and hardening of <em>Dendrobium palpebrae </em>Lindl.Caption</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"Table-1\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1590167174-table1/\">Table-1</a><strong>Table 1. </strong> Development of multiple shoot buds from pseudobulb explant of <em>D. palpebrae </em>when grown on 0.8% (w/v) agar solidified MS media supplemented with different PGRs.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Elongation of multiple shoot buds (MSBs)</strong><br />\r\nThen the multiple shoot buds developed from pseudobulb segment culture were put on elongation media. Eighteen different kinds of solid & liquid media with various combinations and concentrations of PGRs were used for the purpose. Of these, nine were prepared using MS basal medium and the rest nine were prepared using PM basal medium for enhancing elongation of MSBs. In liquid media no agar was added. The efficiency of a medium in terms of enhancing shoot elongation was determined based on the increase in length of shoot system within 30d of culture. Different hormone combinations and culture condition were found to be better for elongation of multiple shoot buds originated tiny plantlets (<a href=\"#Table-2\">Table 2</a>). Highest individual shoot bud increased length was recorded on agar solidified MS medium with 1.0 mg/l Picloram + 1.0 mg/l BAP (3.76 ± 0.14 cm; <a href=\"#figure1\">Figure 1b</a>) followed by liquid MS + 0.5 mg/l NAA + 1.0 mg/l BAP (3.11 ± 0.12 cm; <a href=\"#figure1\">Figure 1c</a>) medium.</p>\r\n\r\n<div id=\"Table-2\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1590167174-table2/\">Table-2</a><strong>Table 2. </strong>Elongation of multiple shoot buds developed from pseudobulb explant of <em>D. papebrae</em> on 0.8% (w/v) agar solidified and liquid media with different kinds of PGRs.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Rooting of individuals shoot bud</strong><br />\r\nThe elongation media are not good enough for root development of young seedlings. So, the developed seedlings were put on rooting media in order to create strong and stout root system. Half strength MS0 and nine different types of PGRs (IAA, IBA, NAA) supplemented MS media were used for induction of strong and stout root system (<a href=\"#Table-3\">Table 3</a>). The efficiency of the rooting media was evaluated based on the increase in length and number of roots developed per seedling within 30d of culture. Increased in length as well as the number of roots developed shoot bud derived seedlings were more on MS medium supplemented with 3% (w/v) sucrose + 0.5 mg/l NAA (4.82 ± 0.22 cm/shoot bud and 2.75 ± 0.17 no/shoot bud and <a href=\"#figure1\">Figure1d</a>) followed by MS with 0.5 mg/l IBA supplemented medium (4.49 ± 0.12 cm/shoot bud and 2.57 ± 0.08 no/shoot bud).</p>\r\n\r\n<div id=\"Table-3\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1590167174-table3/\">Table-3</a><strong>Table 3. </strong>Mean increase in length (cm) and number of roots of <em>D. palpebrae</em> shoot bud originated seedlings* in half strength MS0 and auxin-supplemented MS rooting media.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Development of shoot primordia like structures (SPSs)</strong><br />\r\nMass scale propagation was also done with the use of shoot primodia like structures (SPSs) of D. palpebrae. When the masses of the in vitro grown tiny seedlings of this species were subcultured in MS & PM media both solidified and liquid condition supplemented with different kinds of PGRs those produced SPSs at the base. These SPSs were used for mass scale production of seedlings (<a href=\"#figure1\">Figure 1e</a>).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Hardening and transplantation</strong><br />\r\nThe well-developed plantlets were transferred from culture room to the outside environment through successive phase of acclimatization. Sixty-four per cent of the in vitro grown seedlings survived and continued to grow in pots in the green house. Then, they were finally established in Orchidarium of the Botanical Garden of Chittagong University (<a href=\"#figure1\">Figure 1f</a>).</p>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>Because of long lasting property of <em>Dendrobium palpebrae </em>flowers are commonly used as cut flowers and for that quality these orchids are now used commercially. Some countries such as Thailand, Singapore and Malaysia took the advantage of mass production of commercially important orchids by tissue culture and earn a good amount of foreign currency by exporting it to other countries [<a href=\"#r-12\">12</a>].<br />\r\nMoreover, source of explants, size of explants, media composition, pH and other environmental factors may play a significant role in mass scale clonal propagation of orchids [<a href=\"#r-8\">8</a>]. For micropropagation, noted that lower part of pseudobulb segments showed better response than upper part [<a href=\"#r-13\">13-14</a>]. The cytokines, like BAP and Kn; auxins like IAA, IBA, NAA and Picloram are involved in the process [<a href=\"#r-12\">12</a>]. The requisite of auxins and cytokinins for regeneration of multiple shoot buds and seedlings development has been find out of many orchid species [<a href=\"#r-15\">15-17</a>]. On the other hand, the combinations, concentrations and media type are usually critically important [<a href=\"#r-18\">18</a>]. The ratio of auxin and cytokinins for shoot bud formation varies from species to species. BAP was best for shoot bud formation in <em>Cymbidium gigantean</em>, <em>Vanda spathulata</em> and <em>Dendrobium bensoniae </em>respectively [<a href=\"#r-17\">17-19</a>]<em>.</em><br />\r\nIn elongation of multiple shoot buds (MSBs) was better in agar solidified media than liquid condition. Further MS was found better than PM for elongation of shoot bud [<a href=\"#r-11\">11</a>, <a href=\"#r-20\">20-25</a>]. The elongation rate was different depending on PGR supplements liquid and solid media and solid culture was best for elongation [<a href=\"#r-26\">26-27</a>].<br />\r\nFor induction of strong and stout rooting, auxin supplemented medium was more efficient [<a href=\"#r-28\">28-29</a>]. In some cases, IBA was effective for rooting [<a href=\"#r-30\">30</a>]. Combined effect of BAP (0.5 mg/l) and IBA (1.0 mg/l) was more effective of induced rooting in <em>Dendrobium bensoniae</em> [<a href=\"#r-19\">19</a>]. It is noted that low concentration of auxin is more suitable than high concentration for induction of well-developed root system<em>.</em><br />\r\nLiquid media were more effective in SPSs induction. Comparison of the efficiency of media combination in terms of SPSs induction revealed that liquid media were more effective than agar solidified media. It was further revealed that MS medium was more effective than PM medium for SPSs induction [<a href=\"#r-25\">25-26</a>, <a href=\"#r-31\">31-32</a>].</p>"
},
{
"section_number": 5,
"section_title": "CONCLUSIONS",
"body": "<p>Lower part of pseudobulb segments showed better response than the upper part. MS media supplemented with PGRs like NAA, BAP and Picloram were very effective for MSBs and SPSs development. In elongation media, agar solidified media were better than liquid condition and PGRs supplemented MS media were more effective than PM media. NAA containing MS rooting media were more effective than half strength MS0 and IAA, IBA supplemented full strength MS media. This micropropagation technique can be used by commercial firm for mass scale seedlings production to fulfill demand in international floriculture market<em>.</em></p>"
},
{
"section_number": 6,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>This research received no external funding. We express our heartiest sense of gratitude to Director, National Botanical Garden, Mirpur, Dhaka, Bangladesh for supplying capsules of Dendrobium palpebrae. Specially thanks to the Laboratory of Plant Tissue Culture and Biotechnology, Department of Botany, University of Chittagong, Bangladesh for providing all laboratory facilities during our research.</p>"
},
{
"section_number": 7,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>TKB and MMR were involved in conception and design of the experiments. TKB contributed to perform the experiments. TKB analyzed data. TKB contributed to drafting the article. MMR contributed to revising it critically for important intellectual content. TKB and MMR made the final approval of the version to be published.</p>"
},
{
"section_number": 8,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/37/14/178-1590167174-Figure1.jpg",
"caption": "Figure 1. (1a-1f): In vitro micropropagation, shoot bud elongation, rooting, SPSs development and hardening of Dendrobium palpebrae Lindl.",
"featured": false
}
],
"authors": [
{
"id": 888,
"affiliation": [
{
"affiliation": "Department of Botany, University of Chittagong, Chittagong-4331, Bangladesh."
}
],
"first_name": "Tapash Kumar",
"family_name": "Bhowmik",
"email": "tapashptc@gmail.com",
"author_order": 1,
"ORCID": null,
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Tapash Kumar Bhowmik, Department of Botany, University of Chittagong, Chittagong-4331, Bangladesh. Email: tapashptc@gmail.com",
"article": 201
},
{
"id": 889,
"affiliation": [
{
"affiliation": "Department of Botany, University of Chittagong, Chittagong-4331, Bangladesh."
}
],
"first_name": "Md. Mahbubur",
"family_name": "Rahman",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 201
}
],
"views": 1096,
"downloads": 123,
"references": [
{
"id": 6774,
"serial_number": 1,
"pmc": null,
"reference": "Dressler Phylogeny and classification of the orchid family. Cambridge University Press: Cambridge. 1993.",
"DOI": null,
"article": 201
},
{
"id": 6775,
"serial_number": 2,
"pmc": null,
"reference": "Huda MK. An updated enumeration of the family orchidaceae from Bangladesh. J. Orchid Soc. India. 2007; 21(1-2): 35-49.",
"DOI": null,
"article": 201
},
{
"id": 6776,
"serial_number": 3,
"pmc": null,
"reference": "Bhowmik TK, Rahman MM. In vitro seed germination and flowering of Dendrobium palpebrae Orchid. Journal of Global Biosciences. 2017; 6(1): 4748-4757.",
"DOI": null,
"article": 201
},
{
"id": 6777,
"serial_number": 4,
"pmc": null,
"reference": "Puchooa D. Comparison of different culture media for the in vitro culture of Dendrobium (Orchidaceae). J. Agril. Biol. 2004; 6(5): 884-888.",
"DOI": null,
"article": 201
},
{
"id": 6778,
"serial_number": 5,
"pmc": null,
"reference": "Martin KP, Madassery J. Rapid in vitro propagation of Dendrobium hybrids through direct shoot formation from foliar explants and protocorm-like bodies. Horticult. 2006; 108(1): 95–99.",
"DOI": null,
"article": 201
},
{
"id": 6779,
"serial_number": 6,
"pmc": null,
"reference": "Arditti Aspects of the physiology of orchids. Adv. Bot. Res. 1979; 7: 421-655.",
"DOI": null,
"article": 201
},
{
"id": 6780,
"serial_number": 7,
"pmc": null,
"reference": "Mondal FM. Orchid cultivation and export. 3rd edition. Surovi Publishing Company Limited, Dhaka, Bangladesh. 2011.",
"DOI": null,
"article": 201
},
{
"id": 6781,
"serial_number": 8,
"pmc": null,
"reference": "Bhowmik TK, Rahman MM. Effect of Different Basal Media and PGRs on In Vitro Seed Germination and Seedling Development of Medicinally Important Orchid Cymbidium aloifolium (L.) Sw. J. Pharmacogn Phytochem. 2017; 6(1): 167-172.",
"DOI": null,
"article": 201
},
{
"id": 6782,
"serial_number": 9,
"pmc": null,
"reference": "Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 1962; 15: 473-497.",
"DOI": null,
"article": 201
},
{
"id": 6783,
"serial_number": 10,
"pmc": null,
"reference": "Arditti Clonal propagation of orchids by means of tissue culture: A manual In: J Arditti (ed.), Orchid Biology: Reviews and Perspectives, I. University Press, Ithaca, New York. 1977; pp. 114-1255.",
"DOI": null,
"article": 201
},
{
"id": 6784,
"serial_number": 11,
"pmc": null,
"reference": "Bhadra SK, Bhowmik TK. Axenic germination of seeds and rhizome-based micropropagation of an orchid Arundina graminifolia (D. Don) Hochr. Bangladesh J. Bot. 2005; 34(2): 59-64.",
"DOI": null,
"article": 201
},
{
"id": 6785,
"serial_number": 12,
"pmc": null,
"reference": "Aktar S, Nasiruddin KM, Hossain K. Effects of different media and organic additives interaction on in vitro regeneration of Dendrobium J Agric Rural Dev. 2008; 6(1 & 2): 69-74.",
"DOI": null,
"article": 201
},
{
"id": 6786,
"serial_number": 13,
"pmc": null,
"reference": "Bhadra SK, Hossain MM. In vitro germination and micropropagation of Geodorum densiflorum (Lam.) Scheltr., an endangered orchid species. Plant Tissue Cult. & Biotech. 2003; 13(2): 165-171.",
"DOI": null,
"article": 201
},
{
"id": 6787,
"serial_number": 14,
"pmc": null,
"reference": "Kauth PJ, Vendrame WA, Kane ME. Micropropagation of Calopogon tuberosus, a native terrestrial orchid, by shoot production from corm explants. In vitro, Dev Biol. 2006; 42: 23-23.",
"DOI": null,
"article": 201
},
{
"id": 6788,
"serial_number": 15,
"pmc": null,
"reference": "Arditti J, Ernst R. Micropropagation of orchids, Wiley & Sons Inc., New York. 1993.",
"DOI": null,
"article": 201
},
{
"id": 6789,
"serial_number": 16,
"pmc": null,
"reference": "Hossain MM, Pathak P. In vitro propagation of Dendrobium aphyllum (Orchidaceae) seed germination to flowering. J. Plant Biochem. and Biotechnol. 2012; 22: 157-167.",
"DOI": null,
"article": 201
},
{
"id": 6790,
"serial_number": 17,
"pmc": null,
"reference": "Hossain MM, Sharma M, Teixeira da SJA, Pathak P. Seed germination and tissue culture of Cymbidium giganteum Wall ex Lindl. Sci. Hortic. 2010; 123: 479-487.",
"DOI": null,
"article": 201
},
{
"id": 6791,
"serial_number": 18,
"pmc": null,
"reference": "Decruse SW, Gangaprasad A, Seeni S, Menon VS. Micropropagation and ecorestoration of Vanda spathulata, an exquisite orchid. Plant Cell Tissue & Organ Cult. 2003; 72: 199-202.",
"DOI": null,
"article": 201
},
{
"id": 6792,
"serial_number": 19,
"pmc": null,
"reference": "Riva SS, Islam A, Hoque In vitro Regeneration and Rapid Multiplication of Dendrobium bensoniae, an Indigenous Ornamental Orchid. The Agriculturists. 2016; 14(2): 24-31.",
"DOI": null,
"article": 201
},
{
"id": 6793,
"serial_number": 20,
"pmc": null,
"reference": "Bhowmik TK, Rahman MM. In vitro seed germination and rhizome based micropropagation of Calanthe densiflora Lindl: An indigenous terrestrial orchid of Bangladesh. International Journal of Botany Studies. 2017; 2(1): 110-116.",
"DOI": null,
"article": 201
},
{
"id": 6794,
"serial_number": 21,
"pmc": null,
"reference": "Barua AK, Bhadra SK. In vitro micropropagation of Cymbidium aloifoliumSW. and Spathoglottis plicata Bl. Plant Tissue Cult. & Biotechnol. 1999; 9(2): 133-140.",
"DOI": null,
"article": 201
},
{
"id": 6795,
"serial_number": 22,
"pmc": null,
"reference": "Bhadra SK, Hossain MM. In vitro germination and micropropagation of Geodorum densiflorum (Lam.) Scheltr., an endangered orchid species. Plant Tissue Cult. & Biotechnol. 2003; 13(2): 165-171.",
"DOI": null,
"article": 201
},
{
"id": 6796,
"serial_number": 23,
"pmc": null,
"reference": "Bhadra SK, Bhowmik TK. In vitro seed germination of Phaius tankervilleae (Banks ex L’herit) Blume. The Chittagong University J. of Science. 2005; 29(1): 123-127.",
"DOI": null,
"article": 201
},
{
"id": 6797,
"serial_number": 24,
"pmc": null,
"reference": "Clayton D, Cribb P. The Genus Calanthe. Natural History Publications (Borneo), Kota Kinabalu, in Association with Royal Botanic Gardens Kew, London. 2013.",
"DOI": null,
"article": 201
},
{
"id": 6798,
"serial_number": 25,
"pmc": null,
"reference": "Hoque MI, Roy AR, Sarker RH, Haque MM. Micropropagation of Cymbidium bicolor through in vitro Plant Tissue Cult. & Biotechnol. 1994; 4(1): 45-51.",
"DOI": null,
"article": 201
},
{
"id": 6799,
"serial_number": 26,
"pmc": null,
"reference": "Watad AA, Gaba V, Alper Y, Levin R. Liquid culture as a route for high efficiency micropropagation. In vitro Dev Biol. 1995; 31(3): 30A Abstract.",
"DOI": null,
"article": 201
},
{
"id": 6800,
"serial_number": 27,
"pmc": null,
"reference": "Malabadi RB, Mulgund SG, Kallappa N. Micropropagation of Dendrobium nobile from shoot tip sections. J. Plant Physiol. 2005; 162: 473-478.",
"DOI": null,
"article": 201
},
{
"id": 6801,
"serial_number": 28,
"pmc": null,
"reference": "Banerjee B, Mandal AB. In vitro germination of immature Cymbidium seeds for rapid propagation of plantlets in lands. Cell and Chromo. Res. 1999; 21: 1-5.",
"DOI": null,
"article": 201
},
{
"id": 6802,
"serial_number": 29,
"pmc": null,
"reference": "Gangaprasad AN, Decruse WS, Seeni S, Menon S. Micropropagation and restoration of the endangered Malabar daffodil orchid Ipsea malabarica. Lindleyana. 1999; 14(1): 38-48.",
"DOI": null,
"article": 201
},
{
"id": 6803,
"serial_number": 30,
"pmc": null,
"reference": "Pant B, Swar S. Micropropagation of Cymbidium iridioides Don. J. Orchid Soc. India. 2011; 25(1-2): 9-12.",
"DOI": null,
"article": 201
},
{
"id": 6804,
"serial_number": 31,
"pmc": null,
"reference": "Dasgupta P, Al-Forkan M, Bhadra SK. Micropropagation through seed and shoot segments in Dendrobium crepidatum Plant Tissue Cult. & Biotechnol. 1998; 8(1): 1-10.",
"DOI": null,
"article": 201
},
{
"id": 6805,
"serial_number": 32,
"pmc": null,
"reference": "Roy J, Banerjee N. Induction of callus and plant regeneration from shoot-tip explants of Dendrobium fimbriatum var. oculatum Hk. f. Sci. Horticult. 2003; 97(3-4): 333-340.",
"DOI": null,
"article": 201
}
]
},
{
"id": 60,
"slug": "178-1592221000-self-confidence-as-an-immune-modifying-psychotherapeutic-intervention-for-covid-19-patients-and-understanding-of-its-connection-to-cns-endocrine-immune-axis",
"featured": false,
"slider": false,
"issue": "Special Issue",
"type": "editorial_article",
"manuscript_id": "178-1592221000",
"recieved": "2020-05-17",
"revised": null,
"accepted": "2020-06-14",
"published": "2020-06-16",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/38/178-1592221000.pdf",
"title": "Self-confidence as an immune-modifying psychotherapeutic intervention for COVID-19 patients and understanding of its connection to CNS-endocrine-immune axis",
"abstract": "",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(4): 14-17.",
"academic_editor": "Md. Masudur Rahman, PhD;\r\nSylhet Agricultural University, Sylhet 3100, Bangladesh.",
"cite_info": "Hannan MA, Islam MN, Uddin MJ. Self-confidence as an immune-modifying psychotherapeutic intervention for COVID-19 patients and understanding of its connection to CNS-endocrine-immune axis. J Adv Biotechnol Exp Ther. 2020; 3(4): 14-17.",
"keywords": [],
"DOI": "10.5455/jabet.2020.d151",
"sections": [
{
"section_number": 1,
"section_title": "EDITORIAL",
"body": "<p><strong>C</strong>oronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) not only costs millions of human lives across the globe but also causes uncountable damage to mental health. As the incidence of COVID-19 continues to rise, so does the psychological burden. COVID-19 patients often encounter a variety of psychological stresses, including occupational damage, uncertain future of the family, and even fear of death which cause anxiety and distress, compromising the immune system, and influencing the severity of the disease. Beyond this, stressed individuals are more likely to practice unhealthy habits such as indifference to sleep and food that put them at greater risk.<br />\r\nThe CNS (central nervous system)-endocrine-immune axis is known to be crucially implicated in the stress-mediated dysregulation of the immune response [<a href=\"#r-1\">1</a>]. Following CNS stimulation, stressors provoke the release of several hormones, including catecholamines (epinephrine and norepinephrine), adrenocorticotropin (ACTH), cortisol, growth hormone, and prolactin. Excess secretion of these stress hormones results in overactivation of immune cells and dysregulation of an immune response. Moreover, this interaction between the CNS and the immune system is bidirectional [<a href=\"#r-1\">1</a>]. For example, cytokines (such as IL-1) produced by immune cells stimulate the hypothalamus to release corticotrophin-releasing hormones which further trigger secretion of stress hormones, resulting in immune malfunction.<br />\r\nHowever, interventions that can alleviate psychological stress may control an acute immune response. Self-confidence, a function of mental strength and positive attitude of an individual, is such an intervention that may hold a potential capacity to control stress-mediated immune dysfunction. Several factors, including psychological counseling, social supports, spiritual connections, food habit, health supports, control on emotions, positive thoughts, and reasons for living may help develop self-confidence. Substantial evidence suggests an association between psychological well-being and immune response [<a href=\"#r-2\">2</a>]. It has been observed that psychological intervention was associated with an increase in immune responses by natural killer cells and T cells [<a href=\"#r-3\">3</a>]. Zhao et al, (2016) found that enhancing immune response as indicated by levels of CD4+, CD8+, CD4+ / CD8+ ratio, and free cortisol in serum were involved with the psychological intervention [<a href=\"#r-4\">4</a>]. The level of phagocytes was increased following the psychological intervention [<a href=\"#r-5\">5</a>]. An increase in natural killer (NK) cells after psychological treatments indicates an increase in immune responses [<a href=\"#r-3\">3</a>].<br />\r\nLike psychological intervention, spiritual connections and social supports also boost up the immunity. For instance, an increase in immune cells ( CD4<sup>+</sup>) was observed in patients with IBD [<a href=\"#r-6\">6</a>] and AIDS [<a href=\"#r-7\">7</a>] in response to religious coping and social support. An increase in cortisol response and cortisol awakening response is known to increase immune response in healthy adolescents [<a href=\"#r-8\">8</a>]. Fancourt et al, (2016) showed that music could have a better therapeutic effect on immune responses as was evident by increased levels of cortisol, noradrenaline, beta-endorphin, oxytocin, chemokines and cytokines in cancer patients [<a href=\"#r-9\">9</a>]. Love to the universe, a reason to live may improve the immune system against HIV/AIDS [<a href=\"#r-10\">10</a>]. Religious rituals such as intermittent fasting are recognized to have various health benefits, including boosting up immunity, resistance to stress, and suppressing the aging process [<a href=\"#r-11\">11</a>, <a href=\"#r-12\">12</a>]. In our recent reviews, we proposed fasting and pharmacological intervention by <em>Nigella sativa</em> seed as prospective options against COVID-19 [<a href=\"#r-13\">13</a>, <a href=\"#r-14\">14</a>]. Besides, regular exercise and healthy diets rich in functional ingredients and essential micronutrients will help boost immunity and the anti-stress process.<br />\r\nAlthough self-confidence is a proven quality to succeed in many aspects of our daily life including education, business environments, and politics, it is plausible that confidence could be a vital aspect in combating both communicable (such as COVID-19) and non-communicable diseases/states (such as neurodegenerative diseases and depression). These conditions are often further complicated by patients thinking negatively of their capacities. Using brain imaging studies, it has been revealed that if one thinks positively, “the value areas” of the brain regions including the prefrontal cortex and striatum become activated, leading to the person feeling confident [<a href=\"#r-15\">15</a>]. This process engages brain circuits that are involved in rewards and pleasures so that a person feels better. Taken together, self-confidence may enhance the threshold of vulnerability and prevent further deterioration in COVID-19 infection.<br />\r\nSince there is no specific treatment for COVID-19 yet, the only means to survive from this disease is to consolidate the immune system. In addition, as the immune system is closely linked to the neuroendocrine system, any kind of psychological stresses that overwhelm this association causes immune dysfunction. In this context, self-confidence that can mitigate psychological stresses posed by COVID-19 can play an important role in consolidating immune response against this disease (Figure 1). An in-depth understanding of how psychological stresses overactivate the neuroendocrine-immune axis, weakening the immune system, and how self-confidence interferes with the overwhelmed immune response may offer a prospective strategy for the prevention as well as effective management of COVID-19.</p>\r\n\r\n<figure class=\"image\"><img alt=\"/media/article_images/2023/19/31/Figure_1.jpg\" height=\"84\" src=\"/media/article_images/2023/19/31/Figure_1.jpg\" width=\"100\" />\r\n<figcaption><a href=\"/media/article_images/2023/19/31/Figure_1.jpg\" onclick=\"window.open(this.href, '', 'resizable=no,status=no,location=no,toolbar=no,menubar=no,fullscreen=no,scrollbars=no,dependent=no'); return false;\">Figure 1</a>. Possible mechanisms involved in the impact of self-confidence on COVID-19. Developing self-confidence through psychological interventions, social supports, spiritual connections, food habits, health supports, and positive thinking can potentially reduce psychological stress, thereby consolidating immune functions. Besides, self-confidence can help improve non-communicable diseases such as neurodegenerative as well as inflammatory bowel diseases. As a consequence, self-confidence enables an individual to overcome COVID-19 and reduces the risk of contracting the disease. ACTH, adrenocorticotrophic hormone; GH, growth hormone; NK, natural killer, and APCs, antigen presenting cells.</figcaption>\r\n</figure>"
},
{
"section_number": 2,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>This work is acknowledged to the RP-Grant 2020 of Ewha Womans University, Republic of Korea. MAH acknowledges postdoctoral support from Korea Research Fellowship (KRF) Program (#2018H1D3A1A01074712) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT, Republic of Korea.</p>"
},
{
"section_number": 3,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>This work was a collaboration among all the authors. MJU designed outlines and drafted the manuscript. MAH, MNI, and MJU wrote the initial draft of the manuscript. MJU, MAH, and MNI reviewed the scientific contents described in the manuscript. All authors read and approved the final submitted version of the manuscript.</p>"
},
{
"section_number": 4,
"section_title": "CONFLICT OF INTEREST",
"body": "<p>The authors do not declare any conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2023/19/31/Figure_1.jpg",
"caption": "Figure 1. Possible mechanisms involved in the impact of self-confidence on COVID-19. Developing self-confidence through psychological interventions, social supports, spiritual connections, food habits, health supports, and positive thinking can potentially reduce psychological stress, thereby consolidating immune functions. Besides, self-confidence can help improve non-communicable diseases such as neurodegenerative as well as inflammatory bowel diseases. As a consequence, self-confidence enables an individual to overcome COVID-19 and reduces the risk of contracting the disease. ACTH, adrenocorticotrophic hormone; GH, growth hormone; NK, natural killer, and APCs, antigen presenting cells.",
"featured": true
}
],
"authors": [
{
"id": 197,
"affiliation": [
{
"affiliation": "ABEx Bio-Research Center, Azampur, Dakkhinkhan, Dhaka-1230, Bangladesh"
},
{
"affiliation": "Department of Anatomy, Dongguk University College of Medicine, Gyeongju 38066, Republic of Korea"
},
{
"affiliation": "Department of Biochemistry and Molecular Biology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh"
}
],
"first_name": "Md. Abdul",
"family_name": "Hannan",
"email": null,
"author_order": 1,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 60
},
{
"id": 198,
"affiliation": [
{
"affiliation": "Department of Neuroscience, Yamaguchi University Graduate School of Medicine, 1-1-1, Minami Kogushi, Ube 755-8505, Japan"
}
],
"first_name": "Md. Nabiul",
"family_name": "Islam",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 60
},
{
"id": 199,
"affiliation": [
{
"affiliation": "ABEx Bio-Research Center, Azampur, Dakkhinkhan, Dhaka-1230, Bangladesh"
},
{
"affiliation": "Graduate School of Pharmaceutical Sciences, College of Pharmacy, Ewha Womans University, Seoul, Korea"
}
],
"first_name": "Md Jamal",
"family_name": "Uddin",
"email": "hasan800920@gmail.com",
"author_order": 3,
"ORCID": "https://orcid.org/0000-0003-2911-3255",
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Md Jamal Uddin, PhD; \r\nGraduate School of Pharmaceutical Sciences, College\r\nof Pharmacy, Ewha Womans University, Seoul 03760, Republic of Korea\r\nPhone: +821086737008, Email: hasan800920@gmail.com.",
"article": 60
}
],
"views": 2147,
"downloads": 243,
"references": [
{
"id": 1675,
"serial_number": 1,
"pmc": null,
"reference": "Glaser R, Kiecolt-Glaser JK. Stress-induced immune dysfunction: implications for health. Nat Rev Immunol. 2005;5:243-51.",
"DOI": null,
"article": 60
},
{
"id": 1676,
"serial_number": 2,
"pmc": null,
"reference": "Abdurachman, Herawati N. The Role of Psychological Well-Being in Boosting Immune Response: An Optimal Effort for Tackling Infection. Afr J Infect Dis. 2018;12:54-61.",
"DOI": null,
"article": 60
},
{
"id": 1677,
"serial_number": 3,
"pmc": null,
"reference": "Bakke AC, Purtzer MZ, Newton P. The effect of hypnotic-guided imagery on psychological well-being and immune function in patients with prior breast cancer. J Psychosom Res. 2002;53:1131-7.",
"DOI": null,
"article": 60
},
{
"id": 1678,
"serial_number": 4,
"pmc": null,
"reference": "Zhao X, Cui L, Wang W, Su Q, Li X, Wu J. Influence of psychological intervention on pain and immune functions of patients receiving lung cancer surgery. Pak J Med Sci. 2016;32:155-9.",
"DOI": null,
"article": 60
},
{
"id": 1679,
"serial_number": 5,
"pmc": null,
"reference": "Wu CH, Gau BS. Nursing care of a preschool-age child with cellulites induced by phagocyte deficiency. Hu Li Za Zhi. 2010;57:S16-21.",
"DOI": null,
"article": 60
},
{
"id": 1680,
"serial_number": 6,
"pmc": null,
"reference": "Freitas TH, Hyphantis TN, Andreoulakis E, Quevedo J, Miranda HL, Alves GS, et al. Religious coping and its influence on psychological distress, medication adherence, and quality of life in inflammatory bowel disease. Braz J Psychiatry. 2015;37:219-27.",
"DOI": null,
"article": 60
},
{
"id": 1681,
"serial_number": 7,
"pmc": null,
"reference": "Dalmida SG, Koenig HG, Holstad MM, Wirani MM. The psychological well-being of people living with HIV/AIDS and the role of religious coping and social support. Int J Psychiatry Med. 2013;46:57-83.",
"DOI": null,
"article": 60
},
{
"id": 1682,
"serial_number": 8,
"pmc": null,
"reference": "Rickard NS, Chin TC, Vella-Brodrick DA. Cortisol awakening response as an index of mental health and well-being in adolescents. J Happiness Stud. 2016;17:2555–68.",
"DOI": null,
"article": 60
},
{
"id": 1683,
"serial_number": 9,
"pmc": null,
"reference": "Fancourt D, Williamon A, Carvalho LA, Steptoe A, Dow R, Lewis I. Singing modulates mood, stress, cortisol, cytokine and neuropeptide activity in cancer patients and carers. Ecancermedicalscience. 2016;10:631.",
"DOI": null,
"article": 60
},
{
"id": 1684,
"serial_number": 10,
"pmc": null,
"reference": "SB. Love, Medicine and Miracles. Random House. 1988.",
"DOI": null,
"article": 60
},
{
"id": 1685,
"serial_number": 11,
"pmc": null,
"reference": "Aris JP, Alvers AL, Ferraiuolo RA, Fishwick LK, Hanvivatpong A, Hu D, et al. Autophagy and leucine promote chronological longevity and respiration proficiency during calorie restriction in yeast. Exp Gerontol. 2013;48:1107-19.",
"DOI": null,
"article": 60
},
{
"id": 1686,
"serial_number": 12,
"pmc": null,
"reference": "Rickenbacher A, Jang JH, Limani P, Ungethum U, Lehmann K, Oberkofler CE, et al. Fasting protects liver from ischemic injury through Sirt1-mediated downregulation of circulating HMGB1 in mice. J Hepatol. 2014;61:301-8.",
"DOI": null,
"article": 60
},
{
"id": 1687,
"serial_number": 13,
"pmc": null,
"reference": "Hannan MA, Rahman MA, Rahman MS, Sohag AAM, Das R, Hossian KS, et al. Fasting-mediated priming of host defense against SARS-CoV-2 infection: implication of autophagy and immune response. Preprints. 2020.",
"DOI": null,
"article": 60
},
{
"id": 1688,
"serial_number": 14,
"pmc": null,
"reference": "Islam MN, Hossain MK, Sarker PP, Ferdous J, Hannan MA, Rahman MM, et al. Revisiting pharmacological potentials of Nigella sativa seed: a promising option for COVID-19 prevention and cure. Osfpreprints. 2020.",
"DOI": null,
"article": 60
},
{
"id": 1689,
"serial_number": 15,
"pmc": null,
"reference": "Bault N, Joffily M, Rustichini A, Coricelli G. Medial prefrontal cortex and striatum mediate the influence of social comparison on the decision process. Proc Natl Acad Sci U S A. 2011;108:16044-9.",
"DOI": null,
"article": 60
}
]
},
{
"id": 49,
"slug": "178-1590307525-covid-19-the-catastrophe-of-our-time",
"featured": false,
"slider": false,
"issue": "Special Issue",
"type": "review_article",
"manuscript_id": "178-1590307525",
"recieved": "2020-05-07",
"revised": null,
"accepted": "2020-06-12",
"published": "2020-06-15",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/58/178-1590307525.pdf",
"title": "COVID-19: The catastrophe of our time",
"abstract": "<p>The most discussed topic in today’s world is COVID-19, an acute respiratory disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) because of its contagious transmission pattern, and morbimortality. The virus was originated by bats and in December 2019, first spread to humans by unknown intermediate species in Wuhan, China. The dramatic acceleration of the occurrence and death toll of COVID-19 with no potential medicine and vaccine are enough to explain its severity. This review summarizes multidisciplinary aspects of COVID-19, including origin, epidemiology, symptoms, transmission, pathogenicity, impact on world economy and advances in the use of modern diagnostic procedures and methods. Further, we analyzed extensively for various therapeutic strategies, potential drug options with prospective vaccine candidates and challenges along the way. All data were accumulated through extensive study of recent peer-reviewed publications and authentic reports until June 7, 2020. Collectively, this review would help to shed light on different dimensions of this ongoing pandemic.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(4): 01-13.",
"academic_editor": "Md. Masudur Rahman, PhD\r\nSylhet Agricultural University\r\nBangladesh.",
"cite_info": "Sheam MM, Syed SB, Barman SK, et al. COVID-19: The catastrophe of our time. J Adv Biotechnol Exp Ther. 2020; 3(4): 01-13.",
"keywords": [
"SARS-CoV-2",
"COVID-19",
"Prospective vaccines",
"Therapeutics",
"Economic impact.",
"Pathogenesis"
],
"DOI": "10.5455/jabet.2020.d150",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>The whole world is silenced, and the streets get cleansed in a matter of brief period, and the name responsible for such act is 2019 novel coronavirus (2019-nCoV) or the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). After the emergence of COVID-19 on December 31, 2019, in Wuhan, this outbreak hit all 30 EU/EEA countries and the United Kingdom (UK) within March 15, 2020 [<a href=\"#r-1\">1</a>]. In course of time, April 6, 2020, Spain, Italy, the United States, France and the United Kingdom have been infected catastrophically [<a href=\"#r-2\">2</a>]. According to worldometer (https://www.worldometers.info/coronavirus/), this pandemic spread over 215 countries and territories by June 7, 2020, and confirmed death cases were 402,564 where over 70,000 people died by last two weeks which indicate its ferocity [<a href=\"#r-3\">3</a>]. Developed countries like the United States are now in the shortage of personal protective equipments (PPE) including gloves, face shields, gowns, and hand sanitizer for frontline healthcare workers and high rates of infection are observed among healthcare associates in Italy [<a href=\"#r-4\">4</a>]. As there is no preventive method, lack of intensive care unit (ICU), ventilator, and diagnosing availability could simply raise the mortality rate radically [<a href=\"#r-5\">5</a>].<br />\r\nThe origin of the pandemic is still hypothesized, though the genomic identity of coronavirus isolated from human, and the horseshoe bat <em>Rhinolophus affinis </em>showed enormous similarities [<a href=\"#r-6\">6</a>]. It is reported that human is the only reservoir of the virus. Aside from droplet and contact transmission, some cases suggest aerosol and fecal-oral transmission which needs further verification [<a href=\"#r-7\">7, 8</a>]. After the transmission of SARS-CoV-2, it generally takes 2-14 days for the viral incubation, and symptoms are expressed afterward [<a href=\"#r-9\">9</a>]. As the incubation period, is asymptomatic, the affected person can transmit virus without knowing, which is one of the major reasons for COVID-19 being a pandemic. This evidence of transmission by the asymptomatic carrier has already been found in a case study, described a 20-year-old woman from Wuhan, China, who passed the coronavirus to five family members but never got physically sick herself [<a href=\"#r-10\">10</a>].<br />\r\nIn recent times, the world has reintroduced with some concepts like social/physical distancing, quarantine, lockdown and isolation. As there is yet no validated preventive method, social/physical distancing is the best possible way to limit the infection rate. Through locking down people and contact tracing the infected ones, necessary steps can be taken and that’s why rapid diagnosis in mass scale is a necessity [<a href=\"#r-11\">11</a>]. Though several molecular and serological assays are developed as diagnostics, shortage of reagents, lack of availability of diagnosis, the rapidness of result generation and accuracy of the generated result are creating chaos and anarchy [<a href=\"#r-12\">12</a>].<br />\r\nUntil now, there is no Food and Drug Administration (FDA) licensed drugs for the treatment of COVID-19, but in recent times several drugs, for example, Chloroquine, Remdesivir, Lopinavir–Ritonavir have received attention [<a href=\"#r-13\">13</a>]. Despite the multifaceted challenges, several pharmaceuticals, institutions and universities have embarked on the development of vaccines against SARS-CoV-2, the best possible way to combat this pandemic [<a href=\"#r-14\">14</a>].<br />\r\nGiven the current circumstances, the scientific community is acting rapidly to bring an end to this pandemic. This review aims to provide evidence of early findings, epidemiology, transmission, pathogenesis, risk factors, diagnosis, economic impact as well as suggesting potential drug options, challenges and prospects to develop the COVID-19 vaccine. Additionally, current and upcoming research aspects have also been discussed.</p>"
},
{
"section_number": 2,
"section_title": "ORIGIN AND NAMING",
"body": "<p>The world has witnessed three pandemic outbreaks by the members of the family ‘Coronaviridae’ in the past two decades. In November 2002, first Severe Acute Respiratory Syndrome (SARS) by SARS-Coronavirus (SARS-CoV) had been identified in Guangdong province, Southern China. SARS outbreak continued till July 2003 and there were 8098 confirmed and 774 deaths. The second outbreak, Middle East Respiratory Syndrome (MERS) by MERS-CoV, was first reported in a 60-year-old Saudi Arabian citizen in 2012. MERS was spread to 27 countries and there were confirmed cases and deaths were 2494 and 858, respectively (about 1 in 3). Asian civet cat (<em>Paguma larvata</em>) and dromedary camels were the animal reservoir of SERS and MERS, respectively which is shown in<a href=\"#figure1\"> Figure 1</a>. Recently, the last and deadliest outbreak by the <em>betacoronavirus </em>genus is the COVID-19 by a novel coronavirus where bat is suspected as the zoonotic reservoir. It was first identified in December 2019 in Wuhan, China and WHO declared it as a “Public Health Emergency of International Concern” on 1<sup>st</sup> February 2020 [<a href=\"#r-15\">15</a>] [<a href=\"#r-16\">16</a>]. Till the writing of this article, 215 countries, areas or territories have been affected with 6,999,124 confirmed cases and 402,564 confirmed deaths [<a href=\"#r-3\">3</a>].<br />\r\nIt is presumed that the natural host of SARS-CoV-2 may be the bat <em>Rhinolophus affinis</em>, as the genome nucleotide sequence of coronavirus detected from bat has 96.2% whole genome identity with 2019-nCoV [<a href=\"#r-6\">6</a>]. However, the genetic distance suggests an intermediate in between, which lately presumed as Malayan pangolins. After analyzing 1000 metagenomic samples of pangolins, it was found that 70% of them contained β-CoV and some genome sequence shows maximum 99.92% similarity with the 2019-nCoV [<a href=\"#r-17\">17</a>].<br />\r\nInitially, the newly recognized human pathogen was named as “2019-nCoV” by WHO on January 12, 2020. Later on, the Coronavirus Study Group (CSG) of the International Committee on Virus Taxonomy (ICVT) suggested the name as SARS-CoV-2 as the placement of 2019-nCoV on February 11, 2020 [<a href=\"#r-18\">18</a>, <a href=\"#r-19\">19</a>].</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"625\" src=\"/media/article_images/2024/56/08/178-1590307525-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1.</strong> Origin and transmission model of coronavirus.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 3,
"section_title": "TRANSMISSION AND PATHOGENESIS",
"body": "<p>According to the latest guidelines from Chinese Health Authorities, SARS-CoV-2 generally transmits from person to person via 3 main routes: 1) droplets transmission, 2) contact transmission and 3) aerosol transmission [<a href=\"#r-20\">20</a>]. Infection transmitting droplets varies into different sizes: respiratory droplets are >5-10 µm in diameter, where droplets ranged ≤5μm in diameter is named droplet nuclei. Droplet transmission occurs when a healthy individual is in close contact (1 meter) to COVID-19 affected persons and his/her mucosae (nose, mouth) or conjunctiva (eyes) get exposed to the infective respiratory droplets (via sneezing and/or coughing). In other way around, droplet nuclei are way lighter and stay in the air for longer periods and can transmit to others who are distant more than 1 meter, which should not be mistaken with airborne transmission. To date, no airborne transmission has yet been reported [<a href=\"#r-8\">8</a>].<br />\r\nDirect or indirect contact happens when a subject touches an infected person, a surface or object contaminated by the virus and afterward touches his/ her mouth, nose or eyes which could transmit the virus [<a href=\"#r-20\">20</a>]. Some studies suggested that, through specific medical setup and by procedure specificity, 2019-nCoV could be airborne through fomites and aerosol [<a href=\"#r-8\">8</a>]. It is also noteworthy that, there is evidence of COVID-19 infection leading to intestinal infection and being present in feces [<a href=\"#r-21\">21</a>]. In this case subjects’ nasopharyngeal testing result came negative, consecutive rectal swabs testing came positive all around, indicating the possibility of fecal-oral transmission [<a href=\"#r-7\">7</a>]. However, the aerosol and fecal transmission routes still need to be further studied and confirmed before drawing public attention.<br />\r\nIt should be noted down that though there had been a case report from The New England Journal of Medicine suggesting 2019-nCoV could be spread asymptomatically [<a href=\"#r-22\">22</a>], Kai Kupferschmidt, a correspondent of Science magazine claimed the case report as flawed with significant proves [<a href=\"#r-23\">23</a>].<br />\r\nAfter entering the host through a different portal of entries, Coronavirus (CoV) enters the host cells through binding cell surface receptor by its spike proteins for its replication and finally causing infection [<a href=\"#r-24\">24</a>]. Angiotensin-converting enzyme 2(ACE2) is the spike protein of 2019-nCoV for the entry [<a href=\"#r-25\">25</a>]. By exploiting the host cell machinery, viral proteins are translated from CoV positive-sense RNA, which further undergoes proteolysis mainly by two proteinases namely, coronavirus main proteinase (3CLpro) and the papain-like protease (PLpro) [<a href=\"#r-26\">26</a>]. CoV replicates its genomic material by a replicase which is RNA-dependent RNA polymerase (RdRP) [<a href=\"#r-27\">27</a>]. The above-mentioned spike, RdRP, PLpro and 3CLpro are the possible logical targets for formulating new therapeutics. Figure 2 illustrates the mechanism of pathogenicity and potential therapeutic development of SARS-CoV-2.</p>"
},
{
"section_number": 4,
"section_title": "RISK FACTORS REGARDING COVID-19",
"body": "<p>Reports from the mainland China suggested that most of the confirmed cases were mild or moderate but some of the cases were severe (14%) and critical (5%) [<a href=\"#r-28\">28</a>]. European centre for disease control (ECDC) stated that the COVID-19 risk is moderate for the general people [<a href=\"#r-29\">29</a>] but very high for the older adults and persons who are suffering from chronic diseases [<a href=\"#r-29\">29</a>] [<a href=\"#r-30\">30</a>]. Older age, cardiovascular disease, diabetes, chronic respiratory disease, hypertension, and cancer were all associated with an increased risk of death. Some studies suggested obesity and smoking can increase the risk of severe illness [<a href=\"#r-31\">31</a>]. A study from China found that the deaths between 40-60 year was 16.8%, more likely be to male and having a comorbidity e.g. diabetes, cardiovascular disease, hypertension, or chronic lung disease while who survived were on average 17 years younger [<a href=\"#r-32\">32</a>]. These results have similarities with many other cases in China [<a href=\"#r-31\">31</a>]. Other risk factors could be lack of ICU, ventilator, sampling and testing materials which could accelerate mortality rate [<a href=\"#r-29\">29</a>]. Healthcare personnel are also at risk as they must work in the clinic and hospital. Twenty percent of responding workers were infected in Italy and some of them died [<a href=\"#r-33\">33</a>]. Some countries are even unable to ensure personal protective equipment (PPE) to the health workers and putting them at a great risk of infection which could lead to death [<a href=\"#r-33\">33</a>].</p>"
},
{
"section_number": 5,
"section_title": "SIGNS AND SYMPTOMS",
"body": "<p>The symptoms of COVID-19 are not specific which could be asymptomatic to death causing severe pneumonia. Centre for disease control and prevention (CDC) declared a wide range of symptoms include fever, cough, shortness of breath, chills, repeated shaking with chills, muscle pain, headache, sore throat and new loss of taste or smell. WHO suggested that the COVID-19 symptoms are usually mild and begin gradually [<a href=\"#r-34\">34</a>]. From the previous knowledge of MERS-CoV virus’s incubation period, CDC suggests that the symptoms might take 2-14 days to appear after the exposure [<a href=\"#r-9\">9</a>]. One of the initial studies of 41 patients in Wuhan, China by Chaolin <em>et al</em>. (2020) noted the most common symptoms of the COVID-19 disease i.e. fever (98%), cough (76%), myalgia or fatigue (44%). Some atypical symptoms included as sputum (28%), headache (8%), hemoptysis (5%) and diarrhea (3%). Dyspnea was observed in almost half of the patients and 63% of patients showed lymphocytopenia [35]. Lai <em>et al.</em> (2020) studied three relatively large-scale data of pneumonia patients (278) caused by SARS-CoV-2 in Wuhan [36]. Here, the most common symptoms were fever (92.8%) followed by cough (69.8%), dyspnea (34.5%), myalgia (27.7%), headache (7.2%) and diarrhea (6.1%). Furthermore, a small number of the patient had rhinorrhea (4.0%) and sore throat (5.1%) [36]. Another study by Wei-jie Guan and colleagues with 1099 laboratory-confirmed cases, stated that 43.8% of patients had fever on admission and 88.7% by the time of hospitalization and 67.8% had cough. Additionally, 83.7% of patients had lymphocytopenia and only 3.8% had diarrhea [<a href=\"#r-37\">37</a>]. One recent study reported a new symptom for COVID-19 as the loss of taste and smell. The authors claimed that 30% of positive patients from South Korea experienced anosmia as primary symptom and there is evidence of anosmia in Germany and ageusia in Italy and Switzerland among COVID-19 patients [<a href=\"#r-38\">38</a>]. Data from eight different research groups were accumulated in<a href=\"#Table-1\"> Table 1</a> for getting a clear prediction of the syndrome of COVID-19<strong>,</strong> where the most frequent symptoms are fever (81.44%) followed by cough (64.79%), dyspnea (31.6%), fatigue (28.71%), and myalgia (27.71%). Other mention-worthy signs and symptoms were headache, diarrhea, sputum production, vomiting and nausea, etc.</p>\r\n\r\n<div id=\"Table-1\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1590307525-1/\">Table-1</a><strong>Table 1. </strong>Summarization of the signs and symptoms of COVID-19 in percentage based on the clinical investigations of eight research teams.</p>\r\n</div>"
},
{
"section_number": 6,
"section_title": "DIAGNOSIS OF COVID-19",
"body": "<p>Even if vaccines and other legit therapeutics establishments are underway, there is currently no Food and Drug Administration (FDA) approved vaccines available to conquer the pandemic. That’s why proper diagnosis in mass-scale could minimize the infestation through case identification, isolation and contact tracing [<a href=\"#r-11\">11</a>]. The diagnosis can be indicated as two types based on their availability, result generation speed and reliability. Different serological assays like Rapid Diagnostic Test (RDT), Enzyme-linked Immunosorbent Assay (ELISA) and Neutralization Assay are vastly being used right now.<br />\r\nCompared to ELISA and Neutralization Assay which takes 1-5 h and 3-5 days, respectively, RDT is the most rapid immunoassay as it takes only 10-30 minutes to generate the result [<a href=\"#r-45\">45</a>]. Zhengtu Li and his team developed a new immunoassay, point-of-care testing, which can detect IgM and IgG simultaneously to detect SARS-CoV-2 from the blood and serum samples within 15 minutes [<a href=\"#r-46\">46</a>].<br />\r\nThe test kit has shown some false-negative results as well. It’s hard to determine when the patient exactly gets infected and elevated IgM antibodies disappear after two weeks, which couldn’t meet the required peak of the IgM antibody. The variation of individual antibody production rate can be another reason for the false-negative result exhibition of COVID-19 patients [<a href=\"#r-46\">46</a>].<br />\r\nRather than immunoassay based diagnosis, different commercial and non-commercial serological testing methods still under development which may show even better and rapid viral detection [<a href=\"#r-11\">11</a>]. Nevertheless, molecular testing using real-time Reverse Transcriptase Polymerase Chain Reaction (rRT-PCR) is the most accurate diagnostic procedure because of its ability to target or identify specific pathogens [<a href=\"#r-11\">11</a>]. The samples are collected from the subject’s respiratory tracts. Though upper respiratory samples (nasopharyngeal and oropharyngeal swabs, nasal aspirates) are broadly recommended, lower respiratory samples (sputum, BAL fluid, and tracheal aspirates) are also tested when patients are showing negative test result from upper respiratory samples but exhibiting productive cough and all other COVID-19 symptoms [<a href=\"#r-12\">12</a>]. RNA is extracted from the samples, which is converted to complementary DNA (cDNA) through the reverse transcription process, followed by specific region amplification through primers. The RdRP gene (RNA-dependent RNA polymerase gene) in the ORF1ab region, the E gene (Envelope protein gene) and the N gene (nucleocapsid protein gene) are discovered as the conserved sequence of SARS-CoV-2 and vastly in use as primers in the detection process [<a href=\"#r-47\">47</a>]<strong>.</strong> The assay can then be configured as a two-target system where one primer is responsible for detecting numerous types of coronaviruses including SARS-CoV-2, whereas the other primer is specified only for SARS-CoV-2 detection. After optimizing the assay conditions by taking rigorous steps to set reagent condition, incubation times and temperatures, the PCR test takes place. To ensure the reliability of the test and to detect experimental failures, controls must be carefully chosen [<a href=\"#r-11\">11</a>].<br />\r\nThough viral nucleic acid RT-PCR testing has become the current preferable diagnostic method for SARS-CoV-2 detection, there are some limitations as well. In this crucial time, this testing procedure requires hours to days to generate results, requires complicated laboratory setup, expensive equipment and trained technicians to execute the process. Aside from these, there had been some false-negative cases of RT-PCR generated results [46]. Because of the shortage and time consumption of RT-PCR, the Hubei Province, China used chest Computed Tomography (CT) scans temporarily for diagnosis, where many X-rays are taken from different angles from the patient’s chest to analyze cross-sectional images. The indications of COVID-19 infection include bilateral and peripheral ground-glass opacities (GGO) and consolidations of the lungs [<a href=\"#r-36\">36</a>].</p>"
},
{
"section_number": 7,
"section_title": "POTENTIAL DRUGS AND SUPPORTIVE ASPECTS",
"body": "<p>Although there are no approved drug therapies, common anti-malarial drugs Chloroquine (CQ) and Hydroxychloroquine (HCQ), developed in 1934, have previously gained considerable attention as a possible drug option for COVID-19. Chloroquine can inhibit the viral entry to the host cell by inhibiting viral binding to cell surface receptors and interfere with the viral replication process. Besides, this broad-spectrum antiviral drug can impede post-translational modification of viral proteins [<a href=\"#r-48\">48</a>]. High concentration of Chloroquine analogs has exhibited anti-viral activity against SARS-CoV-2 (EC50=1.13 μM in Vero E6 cells), HIV, dengue, hepatitis C, chikungunya, influenza, Ebola, SARS and MERS viruses <em>in vitro</em> through inhibiting acidification of endosomes [<a href=\"#r-49\">49</a>] [<a href=\"#r-50\">50</a>]. In China, multicenter trials of Chloroquine phosphate against COVID-19 related pneumonia exhibited effectiveness and safety of this drug and is recommended for the treatment of COVID-19 associated pneumonia. More than ten hospitals in China were involved in these trials and results from more than 100 patients exhibited that this drug was successful in inhibiting the severity of pneumonia, relieving viruses and improving the outcome of lung scanning [<a href=\"#r-51\">51</a>]. In April 2020, a trial on 62 COVID-19 patients with HCQ in the Renmin Hospital of Wuhan University was uploaded in the medRxiv server [<a href=\"#r-52\">52</a>]. In this trial, 31 patients in the control group received standard treatment and 31 patients in HCQ treatment group received additional HCQ sulfate tablet (400mg/d) for five days. 80.6% of the HCQ treatment group exhibited improved pneumonia conditions compared to 54.8% of the control group. Also, four patients of the control group proceeded to severe illness, whereas no patient of HCQ treatment group confronted such a situation. In addition, body temperature recovery and cough attenuation period were significantly decreased in the HCQ treatment group compared to the control group [<a href=\"#r-52\">52</a>]. Though the Food and Drug Administration (FDA) authorized temporary use of CQ and HCQ for the treatment of COVID-19 hospitalized patients, on April 24, 2020 FDA reported the cautions against the use of these two drugs due to the serious heart rhythm problems in COVID-19 patients [<a href=\"#r-53\">53</a>]. The study conducted on 368 COVID-19 patients surprisingly supports the findings of FDA as either HCQ or HCQ with Azithromycin could not lower the breathing complications, but the patients treated with only HCQ elevated mortality [<a href=\"#r-54\">54</a>]. Therefore, it needs further investigations regarding the protective roles of HCQ alone and combined with Azithromycin or other antibiotics and other therapeutic molecules. A recent study published in the Nature Cell Research showed Remdesivir was highly efficient in controlling 2019-nCoV in Vero E6 cells and in a human cell line [<a href=\"#r-49\">49</a>]. In Washington, Remdesivir was administrated to a 35-year-old man, the first case of COVID-19 in the United States. Remdesivir exhibited promising outcomes in that case, however, controlled trials are required [<a href=\"#r-21\">21</a>]. A randomized and controlled trial sponsored by the National Institute of Allergy and Infectious Diseases (NIAID) revealed that Remdesivir treatment resulted in faster recovery of hospitalized advanced COVID-19 patients [<a href=\"#r-55\">55</a>]. In particular, remdesivir treatment group exhibited 31% faster recovery (11 days) time than that of placebo group (15 days) [<a href=\"#r-55\">55</a>]. However, in ten hospitals in Hubei, China, randomized, double-blind, placebo-controlled trial of remdesivir on COVID-19 patients didn’t show expected result. Among 155 remdesivir administrated patients, 102 (66%) exhibited adverse effect and in 18 patients (12%) were stopped early due to the unfavorable effects [<a href=\"#r-56\">56</a>]. The status of pneumonia was surprisingly improved within 24 hours in remdesivir administered COVID-19 patients without specific side effects in USA [<a href=\"#r-57\">57</a>,<a href=\"#r-21\">21</a>]. Lopinavir showed <em>in vitro </em>inhibitory activity against SARS-CoV and MARS-CoV and a combination of lopinavir–ritonavir with ribavirin (a guanosine analogue) reduced the death risk in SARS patients. The actions of ribavirin plus both intereferon-α and ciclesonide, a glucocorticoid drug (prescribed for asthma) were also considered as SARS-CoV-2 therapeutic option [<a href=\"#r-58\">58</a>].<br />\r\nAn RdRp inhibitor, favipiravir was used to treat Ebola, emerging influenza in Japan and most recently COVID-19 in China [<a href=\"#r-59\">59</a>, <a href=\"#r-60\">60</a>]. However, a clinical trial on 199 SARS-CoV-2 infected patients treated with lopinavir–ritonavir didn’t show any extra differences or benefits [<a href=\"#r-61\">61</a>]. Furthermore, on February 14, 2020, a clinical trial of Favipiravir (RNA-dependent RNA polymerase inhibitor) against COVID-19 was initiated by the Clinical Medical Research Center of the National Infectious Diseases and the Third People’s Hospital of Shenzhen showed that this drug had more formidable anti SARS-CoV-2 activity than lopinavir/ritonavir without any remarkable adverse effect [<a href=\"#r-59\">59</a>].<br />\r\nMoreover, Azithromycin and Tocilizumab can be prospective adjuvant therapy for COVID-19 [<a href=\"#r-13\">13</a>]. An antibiotic, Teicoplanin prescribed for Methicillin-resistant <em>Staphylococcus aureus</em> (MRSA) which has been reported to be functional against COVID-19 [<a href=\"#r-62\">62</a>]; [<a href=\"#r-63\">63</a>]. The clinical trials of a wide range of therapeutics such as chloroquine/hydroxychloroquine, Bacillus Calmette- Guérin (BCG) vaccine, recombinant human interferon alpha-1b (rhIFNα) nasal drop, lopinavir/ritonavir, chloroquine plus azithromycin, imatinib, favipiravir, telmisartan, colchicine, aspirin, statin and dexamethasone conducted on thousand cases in many countries like as Spain, Canada, Ireland, South Africa, UK, USA, Zambia, China, Singapore, France, Australia, New Zealand, Nigeria, Pakistan and Brazil have got many primary findings but did not conclude absolute perfect one for COVID-19 treatment [<a href=\"#r-64\">64</a>]. These clinical trial-based findings could not establish publicly accepted protocol as well. The effective antiviral impacts of most the tested drugs at extensive level against SARS, MERS or SARS-CoV-2 were observed or the tested drugs were shown to act as the immunomodulatory actions which are believed to decreases the severe lung inflammation at later stages of COVID-19 infection [<a href=\"#r-65\">65-67</a>]. Co-administration of darunavir and umifenovir showed anti-SARS-CoV-2 effects in patient therapies to whom intensive cares were also provided [<a href=\"#r-66\">66</a>]. These findings stressed the scientific communities for designing other more specific antiviral therapeutics for COVID-19 and also pushed the health care providers for exclusive supports for COVID-19 patients.<br />\r\nIt is reported that the nutritional aspects, together with drug interventions not only mitigate CoV infections but also boost up the immunity of the patients. Vitamins A, B, C, D and E, omega‐3 polyunsaturated fatty acids (PUFA) as well as some metals such as selenium, zinc and iron have been reported to possess protective roles against Coronaviruses [<a href=\"#r-68\">68</a>]. Therefore, the single or combined use of them might be very effective nutritional support to combat against SARS-CoV-2. Also, the immune-enhancers, for example, interferons, intravenous gamma globulin, thymosin α‐1, thymopentin, levamisole, cyclosporine A and Chinese medicine exhibited inhibitory activity against SARS‐CoV, MERS‐CoV and avian infectious bronchitis viruses [<a href=\"#r-68\">68</a>]. Potential therapeutic development sites against SARS-CoV-2 have been illustrated in the <a href=\"#figure2\">Figure 2</a>.</p>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"504\" src=\"/media/article_images/2024/56/08/178-1590307525-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2. </strong>Simplified presentation of possible therapeutic development sites for SARS-CoV-2. (a) If the entry of SARS-CoV-2 can be inhibited into the host cells, it is possible to avoid its infections. Monoclonal antibodies, convalescent plasma might be useful to inhibit/modify the spike proteins, ACE2 of SARS-CoV-2 [<a href=\"#r-69\">69</a>]. (b) Positive RNA strand of the virus is directly translated by ribosomes of the host cells. The translated polypeptides undergo proteolysis mainly by two proteinases namely, proteinase 3CLpro and papain-like pro-tease Plpro. Hence these two proteinases might be targeted to inhibit peptide processing after translation. Lopinavir-ritonavir might be useful in this regard. (c) RNA-dependent RNA polymerase (RdRP) transcribes RNA strand from the viral RNA utilizing host cellular machineries. Therefore, inhibition or inactivation of this reverse transcriptase enzyme might be a good option from therapeutic developmental views. For example, Remdisivir is a possible candidate for reverse transcriptase inhibition.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 8,
"section_title": "CHALLENGES AND PROSPECTS TO DEVELOP COVID-19 VACCINE",
"body": "<p><strong>Challenges</strong><br />\r\nIn this current global outbreak of COVID-19, a potential vaccine is a crying need to prevent and combat the disease. Developing a vaccine for infectious disease is usually very time consuming and requires enormous financial support [<a href=\"#r-70\">70</a>]. Safety is the primary concern in developing drugs or vaccines, so it is not recommended to rush in deploying COVID-19 vaccines without maintaining proper safety measures [<a href=\"#r-71\">71</a>]. There are some challenges in developing the vaccine against COVID-19. For instance, Tang <em>et al.</em> (2020) analyzed genomes of SARS-CoV-2 from 103 patients in China and claimed for finding differences in the genome [<a href=\"#r-72\">72</a>], which added a new concern in vaccine development as we might require different vaccines for different genomes. Also, SARS and MERS vaccine candidates intensified the lung disease directly or through antibody-dependent augmentation at preclinical trials. Moreover, to ensure the highest immune response, the most important and critical part is the optimization of antigen design and there is an argument that whether targeting the whole viral protein or just the receptor-binding domain will be the most suitable approach [<a href=\"#r-73\">73</a>]. Besides, a wide range of non-approved drugs for SARS-CoV-2 are being applied to the infected patients though some were effective some were not. As a result, SARS-CoV-2 is getting exposed to different therapeutic compounds which are providing the opportunities to SARS-CoV-2 to be mutated and to be more virulent to human. Changes in the genetic materials might alter the currently available therapeutic targets such as spike protein, RdRP, the main two proteinases PLpro and 3CLpro of SARS-CoV-2. Early studies reported severe illness of SARS-CoV-2 at more than 60 years [<a href=\"#r-74\">74</a>] but recently severe cases have been found in young age groups too, this might be the result of infection of new SARS-CoV-2 strain emerged due to mutation or other reasonable reasons. Besides, the natures of SARS-CoV-2 have not been studied to clearly in response to geographical location and environment too. Moreover, due to the lack of enough data, it is difficult to conclude that people of which age groups are more susceptible to SARS-CoV-2 infection as SARS-CoV and MERS-CoV are strongly related to host conditions including age, biological sex, and overall health [<a href=\"#r-75\">75</a>].</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Prospects</strong><br />\r\nHowever, according to the landscape of COVID-19 candidate vaccines by WHO, until June 2, 2020, there are ten candidate vaccines at clinical evaluation and 123 vaccine candidates are in preclinical evaluation [<a href=\"#r-14\">14</a>]. The University of Oxford and AstraZeneca researchers are the first to begin phase 3 trial of their non-replicating viral vector vaccine and expecting the outcome by summer, 2020. The scientists are engineering a chimpanzee adenovirus to carry DNA for the spike antigen [<a href=\"#r-76\">76</a>]. Other groups using non-replicating viral vector platform are CanSino Biological Inc and the Beijing Institute of Biotechnology, currently in phase 2 of their clinical trial. RNA (mRNA) based approaches reached to clinical trials by Moderna and National Institute of Allergy and Infectious Diseases (phase 2) and BioNTech, Fosun Pharma and Pfizer (phase 1/2) [<a href=\"#r-14\">14</a>]. Similar to maximum COVID-19 vaccines development approaches, Moderna’s candidate are attempting for training the immune system in order to recognising SARS-CoV-2’s spike protein [<a href=\"#r-76\">76</a>]. Moreover, inactivated vaccines from Wuhan Institute of Biological Products/Sinopharm, Beijing Institute of Biological Products/Sinopharm and Sinovac are in the phase 1/2 clinical evaluation. Clinical trial of a protein subunit vaccine (phase 1/2) and a DNA vaccine (phase 1) are conducting by Novavax and Inovio Pharmaceuticals respectively. Furthermore, Institute of Medical Biology of Chinese Academy of Medical Sciences is in phase 1 of their inactivated vaccines’ clinical evaluation [<a href=\"#r-14\">14</a>].<br />\r\nAmong the preclinical vaccines of COVID-19, approaches based on protein subunit are around one third of total evaluation. Sanofi and GlaxoSmithKline, University of Queensland, University of Alberta and some other teams throughout the world are working on protein subunit approach. Live attenuated vaccine is developing by the Serum Institute of India/ Codagenix and Indian Immunologicals Ltd/Griffith University. Furthermore, some companies are working on nucleic acid-based vaccines, for example Inovio Pharmaceuticals (DNA based), Curevac (mRNA based). Live attenuated vaccine is developing by the Serum Institute of India/Codagenix and Indian Immunologicals Ltd/Griffith University. Furthermore, some companies are working on nucleic acid based vaccines, for example Inovio Pharmaceuticals (DNA based), Curevac (mRNA based) [<a href=\"#r-77\">77</a>]. The development of viral vector vaccines against SARS-CoV-2 is in progress by Tonix Pharma and Janseen Pharmaceuticals by adapting the Horepox virus and AdVac® adenoviral vector platform, respectively [<a href=\"#r-78\">78</a>]. The updated list of vaccines against COVID-19 is available at the Draft Landscape of COVID-19 candidate vaccines by WHO.</p>"
},
{
"section_number": 9,
"section_title": "UPCOMING RESEARCH TO COMBAT COVID-19",
"body": "<p>In course of time much more data are getting available to case vitality rates, transmissibility, and overall natural history of COVID-19 infection [<a href=\"#r-79\">79</a>]. Considering the potential warning of the pandemic COVID-19 infections, researchers and physicians are trying their best to understand the new pathophysiology of this novel coronavirus to invent probable treatment strategies, successful therapeutic agents and vaccines [<a href=\"#r-80\">80</a>]. It can be highlighted that antiviral strategies related to small molecules and biologics focusing on complex molecular interactions engaged with coronavirus infection and replication. The drug-repurposing endeavor reported herein emphasized predominantly on the agents proved to be compelling against SARS-CoV, MERS-CoV and other RNA viruses [<a href=\"#r-81\">81</a>].<br />\r\nThe present investigation of coronavirus related biologics such as therapeutic antibodies, cytokines, and nucleic acid-based therapies specially focused on virus gene expression and different types of vaccines [<a href=\"#r-80\">80</a>]. The raised level of inflammatory cytokines IL-6, IL-2, IL-1β, IL-8, IL-17, IFN-γ, TNF-α, IP10, MCP-1, IL-10 and IL-4 resulted cytokine release syndrome (CRS) which might have influential functions in the pathophysiology of COVID-19 [<a href=\"#r-66\">66</a>]. This cytokine aspect should be considered by the scientific societies for the novel therapeutic development of COVID-19. Another important consideration is reinfection of SARS-CoV-2 which is related to cellular immunity and neutralizing antibodies. Hence designed anti-SARS-CoV-2 vaccines should induce rigid cellular immunity and raise the required level of titer of the neutralizing antibodies to make sure no reinfection in the vaccinated populations [<a href=\"#r-82\">82</a>]. Therefore, it is necessary to formulate enzyme-linked immunosorbent assay (ELISA) that enables to quantify anti-receptor binding domain (anti-RBD) antibodies and its correlation with neutralizing antibodies. To generate SARS-CoV specific and neutralizing human monoclonal antibodies (hmAbs), several methods have been applied, for instance, transgenic mice immunization, small chain variable regions’ cloning from immature and convalescent patients as well as convalescent B cells’ immortalization [<a href=\"#r-83\">83</a>].<br />\r\nA study on 129 COVID-19 confirmed cases at mild stage in Wuhan Union Hospital of China suggested the antiviral combined therapies at early stage of infection as it took 7 days less to eradicate viruses compared to late stage of infected cases [<a href=\"#r-84\">84</a>]. Zuo <em>et al.</em> recently demonstrated the shortened SAR-CoV-2 shedding period by administering combined actions of lopinavir/ritonavir plus IFN-α [<a href=\"#r-85\">85</a>]. So, it is another window to carry on research for the development of drugs/vaccines considering the patient physiological situations at early stage of COVID-19 infections. In addition, low dose radiation (1.5 Gy) to COVID-19 pneumonia patient’s both lungs for 10-15 minutes resulted in the improved breathing and recovery on an average 1.5 days [<a href=\"#r-86\">86</a>].<br />\r\nIn current situation, virophage (virus eater) might be a potential research option to destroy this type of notorious virus. It is mention-worthy, because of the emergence of multidrug-resistant bacterial infections, phage therapy has undergone a renaissance [<a href=\"#r-87\">87</a>]. Virophages are double-stranded DNA virus that can infect other viruses and Sputnik was the first isolated virophage which was isolated in 2008 [<a href=\"#r-88\">88,89</a>]. Though virophage has a host range for large-size viruses, future research may isolate prospective virophage with a host range of SARS-CoV-2.</p>"
},
{
"section_number": 10,
"section_title": "IMPACT OF COVID-19 ON GLOBAL ECONOMY",
"body": "<p>The pandemic is playing a devastating role to push the global economy to the verge of great threat, affecting all aspects of economy. According to the United Nations Conference on Trade and Development (UNCTAD) reports, Covid-19 is likely to cost the global economy a minimum of $2 trillion in 2020 [<a href=\"#r-90\">90</a>]. China, where the SARS-Cov-2 virus originated in, is the world’s most populous country and the world’s second biggest economy with a GDP of $13.6 trillion. The country had been fighting the pandemic since December 2019 and taking a great toll in Gross Domestic Product (GDP) which fall to 4.9% in 2020, compared to 6.1% in 2019. The Organisation for Economic Co-operation and Development (OECD) announced that growth prospect for United States of America (USA) the world’s biggest economy, lowered to 1.9% this year after 2.3% in 2019. Consequently, global economic growth is predicted to decline to 2.4% in 2020, compared to 2.9% in 2019 [<a href=\"#r-91\">91</a>].<br />\r\nAccording to the World Travel & Tourism Council (WTTC), tourism is a significant global sector contributing 10.4% of global GDP and 10% of global employments [<a href=\"#r-92\">92</a>]. With the global spread of the virus, the World Tourism Organization (UNWTO) has reported that an estimated downturn in foreign tourist arrivals will be between 20% and 30%, which could result in a drop in international tourism receipts of between 300-450 billion dollars [<a href=\"#r-93\">93</a>]. In view of the deteriorated airline industry, the International Air Transport Association (IATA) estimated losses in the amount of $252 billion and a 44% decrease in lost revenues [<a href=\"#r-94\">94</a>].<br />\r\nGeneral Administration of Customs acknowledged that China, ranked first in goods and services exportation, falls overall exports and imports by 17.2% and 4%, respectively in the first two months of 2020. During that time, industrial production in China dropped by 13.5%, while industrial profit dropped significantly by 38.3% [92]. The International Labour Organization (ILO) has disclosed that the global economic downturn triggered by COVID-19 will eliminate 5.3 to 24.7 million jobs, which means by the end of 2020, a major drop of earnings between $860 billion to $3.4 trillion will happen for the workers [<a href=\"#r-95\">95</a>]. In March 2020, as an example, the unemployment rate in the USA rose by 0.9% to 4.4% [<a href=\"#r-92\">92</a>]. This situation will, in the long term, lead to a deterioration of human assets that has a great effect on countries’ economic development.</p>"
},
{
"section_number": 11,
"section_title": "CONCLUSIONS",
"body": "<p>This study shows not only a universal scenario of the occurrence, distribution, death toll and the aftermath of the current pandemic due to COVID-19 but also the scenario of biomedical research to develop medicine/vaccine. Initially, a lot of studies have been explored based on etiological, epidemiological and diagnostic research. Now research on prevention and control measures have been increasing gradually. Potential studies in this dominion are crying need to control the pandemic soonest possible and to minimize the death toll. The public-private partnership, as well as multinational collaborative research, is essential to eradicate the virus SARS-CoV-2 from the universe like smallpox. This study also recommends the global scientific community and the policymakers for provisioning short-term and long-term public health protection measures to cope up with such type of public health emergency.</p>"
},
{
"section_number": 12,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>We want to acknowledge and grateful to Daniel TA (School of Science, Western Sydney University, Australia) for manuscript language editing service.</p>"
},
{
"section_number": 13,
"section_title": "CONFLICT OF INTEREST",
"body": "<p>The authors do not declare any conflict of interest.</p>"
},
{
"section_number": 14,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>SKB conceived the idea and supervised the project. MMS and SBS performed the database search and literature reviews. SKB, MMS, SBS, SHKB, MRH, RI and DKP wrote the manuscript. SHKB and SBS illustrated the figures. SBS and MMS prepared the table. SKB, DKP and RI critically revised the manuscript. All authors proofread and approved the final manuscript.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/56/08/178-1590307525-Figure1.jpg",
"caption": "Figure 1. Origin and transmission model of coronavirus.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/56/08/178-1590307525-Figure2.jpg",
"caption": "Figure 2. Simplified presentation of possible therapeutic development sites for SARS-CoV-2. (a) If the entry of SARS-CoV-2 can be inhibited into the host cells, it is possible to avoid its infections. Monoclonal antibodies, convalescent plasma might be useful to inhibit/modify the spike proteins, ACE2 of SARS-CoV-2 [69]. (b) Positive RNA strand of the virus is directly translated by ribosomes of the host cells. The translated polypeptides undergo proteolysis mainly by two proteinases namely, proteinase 3CLpro and papain-like pro-tease Plpro. Hence these two proteinases might be targeted to inhibit peptide processing after translation. Lopinavir-ritonavir might be useful in this regard. (c) RNA-dependent RNA polymerase (RdRP) transcribes RNA strand from the viral RNA utilizing host cellular machineries. Therefore, inhibition or inactivation of this reverse transcriptase enzyme might be a good option from therapeutic developmental views. For example, Remdisivir is a possible candidate for reverse transcriptase inhibition.",
"featured": false
}
],
"authors": [
{
"id": 138,
"affiliation": [
{
"affiliation": "Department of Biotechnology and Genetic Engineering, Faculty of Biological Sciences, Islamic University, Kushtia 7003, Bangladesh"
}
],
"first_name": "Md. Moinuddin",
"family_name": "Sheam",
"email": null,
"author_order": 1,
"ORCID": null,
"corresponding": false,
"co_first_author": true,
"co_author": false,
"corresponding_author_info": "",
"article": 49
},
{
"id": 139,
"affiliation": [
{
"affiliation": "Department of Biotechnology and Genetic Engineering, Faculty of Biological Sciences, Islamic University, Kushtia 7003, Bangladesh"
}
],
"first_name": "Shifath Bin",
"family_name": "Syed",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": true,
"co_author": false,
"corresponding_author_info": "",
"article": 49
},
{
"id": 140,
"affiliation": [
{
"affiliation": "School of Science, Western Sydney University, Locked Bag, Penrith NSW, Australia"
}
],
"first_name": "Shital Kumar",
"family_name": "Barman",
"email": null,
"author_order": 3,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 49
},
{
"id": 141,
"affiliation": [
{
"affiliation": "Division of Microbiology, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia"
}
],
"first_name": "Md Rockybul",
"family_name": "Hasan",
"email": null,
"author_order": 4,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 49
},
{
"id": 142,
"affiliation": [
{
"affiliation": "Department of Applied Nutrition and Food Technology, Faculty of Biological Sciences, Islamic University, Kushtia-7003, Bangladesh"
},
{
"affiliation": "Central Laboratory, Islamic University, Kushtia-7003, Bangladesh"
}
],
"first_name": "Dipak Kumar",
"family_name": "Paul",
"email": null,
"author_order": 5,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 49
},
{
"id": 143,
"affiliation": [
{
"affiliation": "Department of Biotechnology and Genetic Engineering, Faculty of Biological Sciences, Islamic University, Kushtia 7003, Bangladesh"
}
],
"first_name": "Rokibul",
"family_name": "Islam",
"email": null,
"author_order": 6,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 49
},
{
"id": 144,
"affiliation": [
{
"affiliation": "Department of Biotechnology and Genetic Engineering, Faculty of Biological Sciences, Islamic University, Kushtia 7003, Bangladesh"
},
{
"affiliation": "Central Laboratory, Islamic University, Kushtia-7003, Bangladesh"
}
],
"first_name": "Sudhangshu Kumar",
"family_name": "Biswas",
"email": "shu_genetics@yahoo.com",
"author_order": 7,
"ORCID": "https://orcid.org/0000-0002-3836-2336",
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Sudhangshu Kumar Biswas, PhD\r\nDepartment of Biotechnology and Genetic Engineering, Faculty of Biological Sciences, Islamic University, Kushtia-7003, Bangladesh, Email: shu_genetics@yahoo.com",
"article": 49
}
],
"views": 1269,
"downloads": 144,
"references": [
{
"id": 7997,
"serial_number": 1,
"pmc": null,
"reference": "Kinross P, Suetens C, Gomes Dias J, Alexakis L, Wijermans A, Colzani E, et al. Rapidly increasing cumulative incidence of coronavirus disease (COVID-19) in the European Union/European Economic Area and the United Kingdom, 1 January to 15 March 2020. Eurosurveillance 2020; 25:1–5.",
"DOI": null,
"article": 49
},
{
"id": 7998,
"serial_number": 2,
"pmc": null,
"reference": "WHO. Coronavirus disease 2019 (COVID-19) Situation Report – 77. 2020.",
"DOI": null,
"article": 49
},
{
"id": 7999,
"serial_number": 3,
"pmc": null,
"reference": "Covid-19 Coronavirus Pandemic. (2020). WorldOmeter. Retrieved June 7, 2020, retrived from https://www.worldometers.info/coronavirus/",
"DOI": null,
"article": 49
},
{
"id": 8000,
"serial_number": 4,
"pmc": null,
"reference": "Ranney ML, Griffeth V, Jha AK. Critical Supply Shortages — The Need for Ventilators and Personal Protective Equipment during the Covid-19 Pandemic. N Engl J Med 2020 (382:e41); 5–7.",
"DOI": null,
"article": 49
},
{
"id": 8001,
"serial_number": 5,
"pmc": null,
"reference": "European center for disease prevention and control. (2020). Coronavirus disease 2019 (COVID-19) pandemic: increased transmission in the EU/EEA and the UK – seventh update, 2019 (March).",
"DOI": null,
"article": 49
},
{
"id": 8002,
"serial_number": 6,
"pmc": null,
"reference": "Peng X, Xu X, Li Y, Cheng L, Zhou X, Ren B. Transmission routes of 2019-nCoV and controls in dental practice. Int J Oral Sci 2020;12:1–6.",
"DOI": null,
"article": 49
},
{
"id": 8003,
"serial_number": 7,
"pmc": null,
"reference": "Xu Y, Li X, Zhu B, Liang H, Fang C, Gong Y, et al. Characteristics of pediatric SARS-CoV-2 infection and potential evidence for persistent fecal viral shedding. Nat Med 2020;26:502-505.",
"DOI": null,
"article": 49
},
{
"id": 8004,
"serial_number": 8,
"pmc": null,
"reference": "WHO. Modes of transmission of virus causing COVID-19 : implications for IPC precaution recommendations. 2020.",
"DOI": null,
"article": 49
},
{
"id": 8005,
"serial_number": 9,
"pmc": null,
"reference": "Symptoms of Coronavirus. (2020). Centers for Disease Control and Prevention (CDC). Retrieved April 8, 2020, from https://www.cdc.gov/coronavirus/2019-ncov/symptoms-testing/symptoms.html",
"DOI": null,
"article": 49
},
{
"id": 8006,
"serial_number": 10,
"pmc": null,
"reference": "Bai Y, Yao L, Wei T, Tian F, Jin D-Y, Chen L, et al. Presumed Asymptomatic Carrier Transmission of COVID-19. J Am Med Assoc 2020;323:1407.",
"DOI": null,
"article": 49
},
{
"id": 8007,
"serial_number": 11,
"pmc": null,
"reference": "Udugama B, Kadhiresan P, Kozlowski HN, Malekjahani A, Osborne M, Li VYC, et al. Diagnosing COVID-19: The Disease and Tools for Detection. ACS Nano 2020; 14: 3822–3835",
"DOI": null,
"article": 49
},
{
"id": 8008,
"serial_number": 12,
"pmc": null,
"reference": "WHO. Laboratory testing for coronavirus disease 2019 (COVID-19) in suspected human cases. Interim Guid 2020:1–7.",
"DOI": null,
"article": 49
},
{
"id": 8009,
"serial_number": 13,
"pmc": null,
"reference": "Smith T, Bushek J, Prosser T. COVID-19 Drug Therapy – Potential Options. Clin Solut 2020.",
"DOI": null,
"article": 49
},
{
"id": 8010,
"serial_number": 14,
"pmc": null,
"reference": "World Health Organization. Draft landscape of COVID-19 candidate vaccines – 2 June, 2020.",
"DOI": null,
"article": 49
},
{
"id": 8011,
"serial_number": 15,
"pmc": null,
"reference": "Peeri NC, Shrestha N, Rahman MS, Zaki R, Tan Z, Bibi S, et al. The SARS, MERS and novel coronavirus (COVID-19) epidemics, the newest and biggest global health threats: what lessons have we learned? Int J Epidemiol 2020:1–10.",
"DOI": null,
"article": 49
},
{
"id": 8012,
"serial_number": 16,
"pmc": null,
"reference": "Wilder-Smith A, Chiew CJ, Lee VJ. Can we contain the COVID-19 outbreak with the same measures as for SARS? Lancet Infect Dis 2020; e102–07",
"DOI": null,
"article": 49
},
{
"id": 8013,
"serial_number": 17,
"pmc": null,
"reference": "Lam TT-Y, Shum MH-H, Zhu H-C, Tong Y-G, Ni X-B, Liao Y-S, et al. Identifying SARS-CoV-2 related coronaviruses in Malayan pangolins. Nature 2020.",
"DOI": null,
"article": 49
},
{
"id": 8014,
"serial_number": 18,
"pmc": null,
"reference": "Jiang S, Shi Z, Shu Y, Song J, Gao GF, Tan W, et al. A distinct name is needed for the new coronavirus. Lancet 2020;395:949.",
"DOI": null,
"article": 49
},
{
"id": 8015,
"serial_number": 19,
"pmc": null,
"reference": "Wang C, Horby PW, Hayden FG, Gao GF. A novel coronavirus outbreak of global health concern. Lancet 2020;395:470–473.",
"DOI": null,
"article": 49
},
{
"id": 8016,
"serial_number": 20,
"pmc": null,
"reference": "Adhikari SP, Meng S, Wu YJ, Mao YP, Ye RX, Wang QZ, et al. Epidemiology, causes, clinical manifestation and diagnosis, prevention and control of coronavirus disease (COVID-19) during the early outbreak period: A scoping review. Infect Dis Poverty 2020;9:1–12.",
"DOI": null,
"article": 49
},
{
"id": 8017,
"serial_number": 21,
"pmc": null,
"reference": "Holshue ML, DeBolt C, Lindquist S, Lofy KH, Wiesman J, Bruce H, et al. First case of 2019 novel coronavirus in the United States. N Engl J Med 2020;382:929–36.",
"DOI": null,
"article": 49
},
{
"id": 8018,
"serial_number": 22,
"pmc": null,
"reference": "Rothe C, Schunk M, Sothmann P, Bretzel G, Froeschl G, Wallrauch C, et al. Transmission of 2019-NCOV infection from an asymptomatic contact in Germany. N Engl J Med 2020;382",
"DOI": null,
"article": 49
},
{
"id": 8019,
"serial_number": 23,
"pmc": null,
"reference": "Kupferschmidt K. Study claiming new coronavirus can be transmitted by people without symptoms was flawed. Science 2020.",
"DOI": null,
"article": 49
},
{
"id": 8020,
"serial_number": 24,
"pmc": null,
"reference": "Li F, Li W, Farzan M, Harrison SC. Structural biology: Structure of SARS coronavirus spike receptor-binding domain complexed with receptor. Science 2005;309:1864–1868.",
"DOI": null,
"article": 49
},
{
"id": 8021,
"serial_number": 25,
"pmc": null,
"reference": "Dong N, Yang X, Ye L, Chen K, Chan EW-C, Chen S. Genomic and protein structure modelling analysis depicts the origin and pathogenicity of 2019-nCoV, a new coronavirus which caused a pneumonia outbreak in Wuhan, China. F1000Research 2020;9:121.",
"DOI": null,
"article": 49
},
{
"id": 8022,
"serial_number": 26,
"pmc": null,
"reference": "Ziebuhr J, Snijder EJ, Gorbalenya AE. Virus-encoded proteinases and proteolytic processing in the Nidovirales. J Gen Virol 2000;81:853–79.",
"DOI": null,
"article": 49
},
{
"id": 8023,
"serial_number": 27,
"pmc": null,
"reference": "Xu X, Liu Y, Weiss S, Arnold E, Sarafianos SG, Ding J. Molecular model of SARS coronavirus polymerase: Implications for biochemical functions and drug design. Nucleic Acids Res 2003;31:7117–30.",
"DOI": null,
"article": 49
},
{
"id": 8024,
"serial_number": 28,
"pmc": null,
"reference": "Wu Z, McGoogan JM. Characteristics of and Important Lessons from the Coronavirus Disease 2019 (COVID-19) Outbreak in China: Summary of a Report of 72314 Cases from the Chinese Center for Disease Control and Prevention. JAMA – J Am Med Assoc 2020; 323:1239-1242.",
"DOI": null,
"article": 49
},
{
"id": 8025,
"serial_number": 29,
"pmc": null,
"reference": "ECDC. Coronavirus disease 2019 (COVID-19) pandemic: increased transmission in the EU/EEA and the UK – seventh update. 2020.",
"DOI": null,
"article": 49
},
{
"id": 8026,
"serial_number": 30,
"pmc": null,
"reference": "Zhou F, Yu T, Du R, Fan G, Liu Y, Liu Z, et al. Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study. Lancet 2020;395: 1054-1062.",
"DOI": null,
"article": 49
},
{
"id": 8027,
"serial_number": 31,
"pmc": null,
"reference": "Jordan RE, Adab P, Cheng KK. Covid-19: risk factors for severe disease and death. Bmj 2020;1198:m1198.",
"DOI": null,
"article": 49
},
{
"id": 8028,
"serial_number": 32,
"pmc": null,
"reference": "Chen T, Wu D, Chen H, Yan W, Yang D, Chen G, et al. Clinical characteristics of 113 deceased patients with coronavirus disease 2019: retrospective study. Bmj 2020;1091:m1091. h",
"DOI": null,
"article": 49
},
{
"id": 8029,
"serial_number": 33,
"pmc": null,
"reference": "Remuzzi A, Remuzzi G. COVID-19 and Italy: what next? Lancet 2020;2:10–3.",
"DOI": null,
"article": 49
},
{
"id": 8030,
"serial_number": 34,
"pmc": null,
"reference": "Coronavirus- Symptoms. (2020). World Health Organization (WHO). Retrieved April 8, 2020, from https://www.who.int/health-topics/coronavirus#tab=tab_3",
"DOI": null,
"article": 49
},
{
"id": 8031,
"serial_number": 35,
"pmc": null,
"reference": "Wu D, Wu T, Liu Q, Yang Z. The SARS-CoV-2 outbreak: what we know. Int J Infect Dis 2020; 94: 44-48.",
"DOI": null,
"article": 49
},
{
"id": 8032,
"serial_number": 36,
"pmc": null,
"reference": "Lai CC, Shih TP, Ko WC, Tang HJ, Hsueh PR. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and coronavirus disease-2019 (COVID-19): The epidemic and the challenges. Int J Antimicrob Agents 2020;55:105924.",
"DOI": null,
"article": 49
},
{
"id": 8033,
"serial_number": 37,
"pmc": null,
"reference": "Guan W, Ni Z, Hu Y, Liang W, Ou C, He J, et al. Clinical characteristics of 2019 novel coronavirus infection in China. MedRxiv 2020.",
"DOI": null,
"article": 49
},
{
"id": 8034,
"serial_number": 38,
"pmc": null,
"reference": "Gautier J-F, Ravussin Y. A New Symptom of COVID-19: Loss of Taste and Smell. Obesity 2020; 28.",
"DOI": null,
"article": 49
},
{
"id": 8035,
"serial_number": 39,
"pmc": null,
"reference": "Wang D, Hu B, Hu C, Zhu F, Liu X, Zhang J, et al. Clinical Characteristics of 138 Hospitalized Patients with 2019 Novel Coronavirus-Infected Pneumonia in Wuhan, China. JAMA – J Am Med Assoc 2020;323:1061–9.",
"DOI": null,
"article": 49
},
{
"id": 8036,
"serial_number": 40,
"pmc": null,
"reference": "Chen N, Zhou M, Dong X, Qu J, Gong F, Han Y, et al. Epidemiological and clinical characteristics of 99 cases of 2019 novel coronavirus pneumonia in Wuhan, China: a descriptive study. Lancet 2020;395:507–13.",
"DOI": null,
"article": 49
},
{
"id": 8037,
"serial_number": 41,
"pmc": null,
"reference": "Song F, Shi N, Shan F, Zhang Z, Shen J, Lu H, et al. Emerging 2019 novel coronavirus (2019-NCoV) pneumonia. Radiology 2020;295:210–7.",
"DOI": null,
"article": 49
},
{
"id": 8038,
"serial_number": 42,
"pmc": null,
"reference": "Huang C, Wang Y, Li X, Ren L, Zhao J, Hu Y, et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet 2020;395:497–506.",
"DOI": null,
"article": 49
},
{
"id": 8039,
"serial_number": 43,
"pmc": null,
"reference": "Lei C, Huiguo L, Wei L, Jing L, Kui L, Jin S, et al. Analysis of clinical features of 29 patients with 2019 novel coronavirus pneumonia. Chinese J Tuberc Respir Dis 2020; 43:E005.",
"DOI": null,
"article": 49
},
{
"id": 8040,
"serial_number": 44,
"pmc": null,
"reference": "Chang D, Lin M, Wei L, Xie L, Zhu G, Cruz CS Dela, et al. Epidemiologic and Clinical Characteristics of Novel Coronavirus Infections Involving 13 Patients Outside Wuhan, China. JAMA – J Am Med Assoc 2020;323:1093.",
"DOI": null,
"article": 49
},
{
"id": 8041,
"serial_number": 45,
"pmc": null,
"reference": "Serology-based tests for COVID-19. (2020). Center for Health Security, John Hopkins University. Retrieved April 8, 2020, retrived from http://www.centerforhealthsecurity.org/resources/COVID-19/Serology-based-tests-for-COVID-19.html",
"DOI": null,
"article": 49
},
{
"id": 8042,
"serial_number": 46,
"pmc": null,
"reference": "Li Z, Yi Y, Luo X, Xiong N, Liu Y, Li S, et al. Development and Clinical Application of A Rapid IgM‐IgG Combined Antibody Test for SARS‐CoV‐2 Infection Diagnosis. J Med Virol 2020;1–7.",
"DOI": null,
"article": 49
},
{
"id": 8043,
"serial_number": 47,
"pmc": null,
"reference": "Corman VM, Landt O, Kaiser M, Molenkamp R, Meijer A, Chu DK, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill 2020;25:1–8.",
"DOI": null,
"article": 49
},
{
"id": 8044,
"serial_number": 48,
"pmc": null,
"reference": "Devaux CA, Rolain J-M, Colson P, Raoult D. New insights on the antiviral effects of chloroquine against coronavirus: what to expect for COVID-19? Int J Antimicrob Agents 2020:105938.",
"DOI": null,
"article": 49
},
{
"id": 8045,
"serial_number": 49,
"pmc": null,
"reference": "Wang M, Cao R, Zhang L, Yang X, Liu J, Xu M, et al. Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro. Cell Res 2020;30:269–71.",
"DOI": null,
"article": 49
},
{
"id": 8046,
"serial_number": 50,
"pmc": null,
"reference": "Al-Bari MAA. Targeting endosomal acidification by chloroquine analogs as a promising strategy for the treatment of emerging viral diseases. Pharmacol Res Perspect 2017;5:1–13.",
"DOI": null,
"article": 49
},
{
"id": 8047,
"serial_number": 51,
"pmc": null,
"reference": "Gao J, Tian Z, Yang X. Breakthrough: Chloroquine phosphate has shown apparent efficacy in treatment of COVID-19 associated pneumonia in clinical studies. Biosci Trends 2020;14:72–73.",
"DOI": null,
"article": 49
},
{
"id": 8048,
"serial_number": 52,
"pmc": null,
"reference": "Chen Z, Hu J, Zhang Z, Jiang S, Han S, Yan D, et al. Efficacy of hydroxychloroquine in patients with COVID-19: results of a randomized clinical trial. MedRxiv 2020; https://doi.org/10.1101/2020.03.22.20040758.",
"DOI": null,
"article": 49
},
{
"id": 8049,
"serial_number": 53,
"pmc": null,
"reference": "FDA cautions against use of hydroxychloroquine or chloroquine for COVID-19 outside of the hospital setting or a clinical trial due to risk of heart rhythm problems. Food Drug Adm 2020.",
"DOI": null,
"article": 49
},
{
"id": 8050,
"serial_number": 54,
"pmc": null,
"reference": "Magagnoli J, Narendran S, Pereira F, Cummings T, Hardin JW, Sutton SS, et al. Outcomes of hydroxychloroquine usage in United States veterans hospitalized with Covid-19. MedRxiv 2020.https://doi.org/10.1101/2020.04.16.20065920.",
"DOI": null,
"article": 49
},
{
"id": 8051,
"serial_number": 55,
"pmc": null,
"reference": "NIH clinical trial shows Remdesivir accelerates recovery from advanced COVID-19. (n.d.). National Institute of Health. Retrieved June 7, 2020, from https://www.nih.gov/news-events/news-releases/nih-clinical-trial-shows-remdesivir-accelerates-recovery-advanced-covid-19",
"DOI": null,
"article": 49
},
{
"id": 8052,
"serial_number": 56,
"pmc": null,
"reference": "Wang Y, Zhang D, Du G, Du R, Zhao J, Jin Y, et al. Remdesivir in adults with severe COVID-19: a randomised, double-blind, placebo-controlled, multicentre trial. Lancet 2020;0:1569–78.",
"DOI": null,
"article": 49
},
{
"id": 8053,
"serial_number": 57,
"pmc": null,
"reference": "Ko W, Rolain J, Lee N, Chen P, Huang C. Arguments in favour of remdesivir for treating SARS-CoV-2 infections. J Antimicrob Agents 2020;55: 105933",
"DOI": null,
"article": 49
},
{
"id": 8054,
"serial_number": 58,
"pmc": null,
"reference": "Development of coronavirus treatment advancing in Japan with existing meds. (n.d.). The Mainichi. Retrieved June 6, 2020, retrived from https://mainichi.jp/english/articles/20200317/p2a/00m/0na/026000c",
"DOI": null,
"article": 49
},
{
"id": 8055,
"serial_number": 59,
"pmc": null,
"reference": "Dong L, Hu S, Gao J. Discovering drugs to treat coronavirus disease 2019 (COVID-19). Drug Discov Ther 2020;14:58–60..",
"DOI": null,
"article": 49
},
{
"id": 8056,
"serial_number": 60,
"pmc": null,
"reference": "Nagata T, Lefor AK, Hasegawa M, Ishii M. Favipiravir: A New Medication for the Ebola Virus Disease Pandemic. Disaster Med Public Health Prep 2015;9:79–81.",
"DOI": null,
"article": 49
},
{
"id": 8057,
"serial_number": 61,
"pmc": null,
"reference": "Cao B, Wang Y, Wen D, Liu W, Wang J, Fan G, et al. A Trial of Lopinavir–Ritonavir in Adults Hospitalized with Severe Covid-19. N Engl J Med 2020:1–13.",
"DOI": null,
"article": 49
},
{
"id": 8058,
"serial_number": 62,
"pmc": null,
"reference": "Ramos-Martín V, Johnson A, McEntee L, Farrington N, Padmore K, Cojutti P, et al. Pharmacodynamics of teicoplanin against MRSA. J Antimicrob Chemother 2017;72:3382–9.",
"DOI": null,
"article": 49
},
{
"id": 8059,
"serial_number": 63,
"pmc": null,
"reference": "Baron SA, Devaux C, Colson P, Raoult D, Rolain JM. Teicoplanin: an alternative drug for the treatment of COVID-19? Int J Antimicrob Agents 2020;55:105944.",
"DOI": null,
"article": 49
},
{
"id": 8060,
"serial_number": 64,
"pmc": null,
"reference": "Davis JS, Ferreira D, Denholm JT, Tong SY. Clinical trials for the prevention and treatment of Coronavirus Disease 2019 (COVID-19): The current state of play. Med J Aust 2020.",
"DOI": null,
"article": 49
},
{
"id": 8061,
"serial_number": 65,
"pmc": null,
"reference": "Yao X, Ye F, Zhang M, Cui C, Huang B, Niu P, et al. In Vitro Antiviral Activity and Projection of Optimized Dosing Design of Hydroxychloroquine for the Treatment of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Clin Infect Dis 2020;2:1–25.",
"DOI": null,
"article": 49
},
{
"id": 8062,
"serial_number": 66,
"pmc": null,
"reference": "Costanzo M, De Giglio MAR, Roviello GN. SARS CoV-2: Recent Reports on Antiviral Therapies Based on Lopinavir/Ritonavir, Darunavir/Umifenovir, Hydroxychloroquine, Remdesivir, Favipiravir and Other Drugs for the Treatment of the New Coronavirus. Curr Med Chem 2020;27.",
"DOI": null,
"article": 49
},
{
"id": 8063,
"serial_number": 67,
"pmc": null,
"reference": "Liu B, Li M, Zhou Z, Guan X, Xiang Y. Can we use interleukin-6 (IL-6) blockade for coronavirus disease 2019 (COVID-19)-induced cytokine release syndrome (CRS)? J Autoimmun 2020;111:102452.",
"DOI": null,
"article": 49
},
{
"id": 8064,
"serial_number": 68,
"pmc": null,
"reference": "Zhang L, Liu Y. Potential interventions for novel coronavirus in China: A systematic review. J Med Virol 2020;92:479–90..",
"DOI": null,
"article": 49
},
{
"id": 8065,
"serial_number": 69,
"pmc": null,
"reference": "Wang C, Li W, Drabek D, Okba NMA, Haperen R van, Osterhaus ADME, et al. A human monoclonal antibody blocking SARS-CoV-2 infection. BioRxiv 2020.",
"DOI": null,
"article": 49
},
{
"id": 8066,
"serial_number": 70,
"pmc": null,
"reference": "Ahn D-G, Shin H-J, Kim M-H, Lee S, Kim H-S, Myoung J, et al. Current Status of Epidemiology, Diagnosis, Therapeutics, and Vaccines for Novel Coronavirus Disease 2019 (COVID-19). J Microbiol Biotechnol 2020;30:313–24.",
"DOI": null,
"article": 49
},
{
"id": 8067,
"serial_number": 71,
"pmc": null,
"reference": "Shibo Jiang. Don’t rush to deploy COVID-19 vaccines and drugs without sufficient safety guarantees. Nature 2020.",
"DOI": null,
"article": 49
},
{
"id": 8068,
"serial_number": 72,
"pmc": null,
"reference": "Tang X, Wu C, Li X, Song Y, Yao X, Wu X, et al. On the origin and continuing evolution of SARS-CoV-2. Natl Sci Rev 2020; 0:1-12.",
"DOI": null,
"article": 49
},
{
"id": 8069,
"serial_number": 73,
"pmc": null,
"reference": "Lurie N, Saville M, Hatchett R, Halton J. Developing Covid-19 Vaccines at Pandemic Speed. N Engl J Med 2020:1–5.",
"DOI": null,
"article": 49
},
{
"id": 8070,
"serial_number": 74,
"pmc": null,
"reference": "Gralinski LE, Menachery VD. Return of the coronavirus: 2019-nCoV. Viruses 2020;12:1–8.",
"DOI": null,
"article": 49
},
{
"id": 8071,
"serial_number": 75,
"pmc": null,
"reference": "Fehr AR, Channappanavar R, Perlman S. Middle East Respiratory Syndrome: Emergence of a Pathogenic Human Coronavirus. Annu Rev Med 2017;68:387–99.",
"DOI": null,
"article": 49
},
{
"id": 8072,
"serial_number": 76,
"pmc": null,
"reference": "Mullard A. World Report COVID-19 vaccine development pipeline gears up. Lancet 2020;395:1751–2..",
"DOI": null,
"article": 49
},
{
"id": 8073,
"serial_number": 77,
"pmc": null,
"reference": "Corey L, Mascola JR, Fauci AS, Collins FS. A strategic approach to COVID-19 vaccine R&D. Science 2020;368:948–50.",
"DOI": null,
"article": 49
},
{
"id": 8074,
"serial_number": 78,
"pmc": null,
"reference": "Zhang J, Zeng H, Gu J, Li H, Zheng L, Zou Q. Progress and Prospects on Vaccine Development against SARS-CoV-2. Vaccines 2020, Vol 8, Page 153 2020;8.",
"DOI": null,
"article": 49
},
{
"id": 8075,
"serial_number": 79,
"pmc": null,
"reference": "Quinn JM, Haggenmiller C, Wilson JM, McNamara T, Goebbels S, Hansen J-C, et al. Conference Report: Global Health Security Alliance (GLoHSA), a Product of the World Health Summit. Disaster Med Public Health Prep 2019:1–3.",
"DOI": null,
"article": 49
},
{
"id": 8076,
"serial_number": 80,
"pmc": null,
"reference": "Liu C, Zhou Q, Li Y, Garner L V., Watkins SP, Carter LJ, et al. Research and Development on Therapeutic Agents and Vaccines for COVID-19 and Related Human Coronavirus Diseases. ACS Cent Sci 2020;6:315-331.",
"DOI": null,
"article": 49
},
{
"id": 8077,
"serial_number": 81,
"pmc": null,
"reference": "Gorbalenya AE, Baker SC, Baric RS, Groot RJ De, Gulyaeva AA, Haagmans BL, et al. Severe acute respiratory syndrome-related coronavirus: The species and its viruses – a statement of the Coronavirus Study Group. BioRxiv 2020.",
"DOI": null,
"article": 49
},
{
"id": 8078,
"serial_number": 82,
"pmc": null,
"reference": "Manners C, Bautista EL, Sidoti H, Lopez OJ. Protective Adaptive Immunity Against Severe Acute Respiratory Syndrome Coronaviruses 2 (SARS-CoV-2) and Implications for Vaccines. Cureus 2020;2:6–13.",
"DOI": null,
"article": 49
},
{
"id": 8079,
"serial_number": 83,
"pmc": null,
"reference": "Coughlin MM, Prabhakar BS. Neutralizing human monoclonal antibodies to severe acute respiratory syndrome coronavirus: target, mechanism of action, and therapeutic potential. Rev Med Virol 2012;22:2–17.",
"DOI": null,
"article": 49
},
{
"id": 8080,
"serial_number": 84,
"pmc": null,
"reference": "Yu T, Tian C, Chu S, Zhou H, Zhang Z, Luo S, et al. COVID-19 patients benefit from early antiviral treatment: a comparative, retrospective study. J Med Virol 2020.",
"DOI": null,
"article": 49
},
{
"id": 8081,
"serial_number": 85,
"pmc": null,
"reference": "Zuo Y, Liu Y, Zhong Q, Zhang K, Xu Y, Wang Z. Lopinavir/ritonavir and interferon combination therapy may help shorten the duration of viral shedding in patients with COVID-19: a retrospective study in two designated hospitals in Anhui, China. J Med Virol 2020.",
"DOI": null,
"article": 49
},
{
"id": 8082,
"serial_number": 86,
"pmc": null,
"reference": "Hess C. B., Buchwald Z. S., Stokes W., Switchenko J. M., Nasti T. H., Weinberg, B. D., Khan M. K. (2020). Low-Dose Whole-Lung Radiation for COVID-19 Pneumonia: Planned Day-7 Interim Analysis of an Ongoing Clinical Trial. medRxiv 2020.",
"DOI": null,
"article": 49
},
{
"id": 8083,
"serial_number": 87,
"pmc": null,
"reference": "Oliveira H, Sillankorva S, Merabishvili M, Kluskens LD, Azeredo J. Unexploited opportunities for phage therapy. Front Pharmacol 2015;6:1–4.",
"DOI": null,
"article": 49
},
{
"id": 8084,
"serial_number": 88,
"pmc": null,
"reference": "Sobhy H. Virophages and their interactions with giant viruses and host cells. Proteomes 2018;6.",
"DOI": null,
"article": 49
},
{
"id": 8085,
"serial_number": 89,
"pmc": null,
"reference": "La Scola B, Desnues C, Pagnier I, Robert C, Barrassi L, Fournous G, et al. The virophage as a unique parasite of the giant mimivirus. Nature 2008;455:100–4.",
"DOI": null,
"article": 49
},
{
"id": 8086,
"serial_number": 90,
"pmc": null,
"reference": "The Coronavirus Shock: A Story of Another Global Crisis Foretold and Policymaker Should Be Doing About It. (n.d.). Geneva, Switzerland.",
"DOI": null,
"article": 49
},
{
"id": 8087,
"serial_number": 91,
"pmc": null,
"reference": "OECD. Coronavirus: The world economy at risk. OECD Interim Econ Assess 2020:1–15.",
"DOI": null,
"article": 49
},
{
"id": 8088,
"serial_number": 92,
"pmc": null,
"reference": "Açikgöz Ö, Günay A. The early impact of the Covid-19 pandemic on the global and Turkish economy. Turkish J Med Sci 2020;50:520–6.",
"DOI": null,
"article": 49
},
{
"id": 8089,
"serial_number": 93,
"pmc": null,
"reference": "International Tourist Arrival Could Fall By 20-30% In 2020. World Tour Organ 2020.",
"DOI": null,
"article": 49
},
{
"id": 8090,
"serial_number": 94,
"pmc": null,
"reference": "IATA Economics. COVID-19 Cash burn analysis. IATA Econ 2020.",
"DOI": null,
"article": 49
},
{
"id": 8091,
"serial_number": 95,
"pmc": null,
"reference": "International Labor Organization. COVID-19 and the world of work: Impact and policy responses 2020:1–15.",
"DOI": null,
"article": 49
}
]
},
{
"id": 200,
"slug": "178-1590260727-biological-investigations-of-the-methanol-extract-of-tetrastigma-leucostaphylum-dennst-alston-ex-mabb-vitaceae-in-vivo-and-in-vitro-approach",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1590260727",
"recieved": "2020-04-07",
"revised": null,
"accepted": "2020-06-12",
"published": "2020-06-14",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/31/178-1590260727.pdf",
"title": "Biological investigations of the methanol extract of Tetrastigma leucostaphylum (Dennst.) Alston ex Mabb. (Vitaceae) : In vivo and in vitro approach",
"abstract": "<p><em>Tetrastigma leucostaphylum </em>(Family: Vitaceae) is popular for its medicinal value in Bangladeshi tribal communities. This study aims to investigate several pharmacological values of methanol extract of <em>T. leucostaphylum</em> (METL). <em>In vivo</em> analgesic and anti-inflammatory researches have been implemented by using acetic acid-induced writhing and formalin-induced paw licking test protocols in mice. Furthermore, <em>in vitro</em> thrombolytic and anthelmintic studies have been performed by following the blood clot lysis method and nematode mortality measurement method. In the <em>in vivo</em> study, METL did not minimize the acetic acid-induced writhes prominently but significantly attenuate both the peripheral and inflammatory pain in mice in a dose-dependent manner. In early and late phase, METL 400 (mg/kg, b.w; p.o) showed 39.63 % and 48.73 % paw licking inhibition. Again, METL (100 µL) reflected 56.62 % clot lysis in thrombolytic research. Besides, METL causes the death of nematodes in a dose-dependent manner. The bioassay of the methanol extract of <em>T. leucostaphylum</em> justified the analgesic, anti-inflammatory, thrombolytic and anthelmintic activities of the crude extract and finally suggests the test extract as a wellspring of anti-inflammatory, thrombolytic and anthelmintic agents as a crude drug source.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 216-224.",
"academic_editor": "Dr. Masud Parvez, Washington State University, USA.",
"cite_info": "Rudra S, Sawon MSU, et al. Biological investigations of the methanol extract of Tetrastigma leucostaphylum (Dennst.) Alston ex Mabb. (Vitaceae) : In vivo and in vitro approach. J Adv Biotechnol Exp Ther. 2020; 3(3): 216-224.",
"keywords": [
"Tetrastigma leucostaphylum",
"Tubifex tubifex",
"Formalin",
"Writhing",
"Clot lysis"
],
"DOI": "10.5455/jabet.2020.d127",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>Pain is a troubling perception often resulting from severe or harmful stimuli [<a href=\"#r-1\">1</a>] where inflammation is the part of a complex physiological reaction to harmful stimuli including pathogenes, injured cells, or irritants by body tissue. It is a defensive response involving immune cells and blood vessels [<a href=\"#r-2\">2</a>]. In spite of conducting several types of researches to develop pain therapy, still, the place of operative and promising analgesic candidates especially for the management of chronic pain is not entirely up for grabs [<a href=\"#r-3\">3</a>]. Phytoconstituents derived from herbal plants are considered as a noteworthy source of novel bioactive molecules exhibiting promising therapeutic activities [<a href=\"#r-4\">4</a>]. Phytochemical studies based on folkloric usage of the medicinal plant to alleviate pain have become rational and promising to discover newer analgesic agents [<a href=\"#r-5\">5</a>]. Alkaloid (morphine), one of the vastly used pain relievers is widespread in treating intense and remorseless pain including injury or cancer-induced pain though it has few severe side effects and thus the attention of pharmaceutical companies is grabbed to evaluate different medicinal plant extractives to bring newer pain killers with less side effect and more specificity to light [<a href=\"#r-6\">6</a>]. Studies, stratified into various types of pain tests reflect remarkable anti-nociception activity achieved by systemic administration of several alkaloids, terpenoids, and flavonoids such as rutin, quercetin, pectolinarin, and gossypin [<a href=\"#r-3\">3</a>]. Intense breakdown of homeostasis like injury, infection and/or exposure to contaminants can provoke innate immune receptors by pointing out the pathogens with a target of eliminating those from bodies is known as inflammation categorized by redness, pain, heat, swelling, and loss of function in the affected zone [<a href=\"#r-7\">7, 8</a>]. In recent practice, both NSAIDs (Nonsteroidal Anti-Inflammatory Drugs) and steroidal drugs are taken under consideration to treat pain and inflammation through the use of NSAIDs for a prolonged period. But it can impose several side effects and organ damage including liver and gastrointestinal tract along with initiation of cardiovascular complications and renal failure [<a href=\"#r-9\">9, 10</a>]. Thus, to alleviate pain bioactive constituents isolated from plants have been inspected for centuries to establish phytochemicals as anti-inflammatory agents [<a href=\"#r-11\">11</a>]. Besides, phytomedicines acquired from plant extracts may satisfy the desire of nontoxic, more potent, fruitful, and safe drugs to cure pain and inflammation more efficiently [<a href=\"#r-8\">8</a>].<br />\r\nThrombosis is a fatal disease categorized by thrombus formation in the circulatory system [<a href=\"#r-12\">12</a>]. Alteplase, anistreplase, streptokinase, urokinase, and tissue plasminogen activator (tPA) are abundantly used anti-thrombus agents that are administered to dissolve blood clot characterized as thrombolysis process [<a href=\"#r-13\">13, 14</a>]. Though aspirin and heparin are safe to use, they are not profoundly effective to lysis clots and to prevent occlusion. Again, Thrombin inhibitors and antiplatelet agents that are more selective and are supposed to be more potent are having dubious safety [<a href=\"#r-15\">15</a>]. Streptokinase and urokinase are frequently used first-generation drugs to treat thrombosis which possess serious side effects including anaphylactic reaction, systemic fibrinolysis, and hemorrhage due to their weak substrate specificity [<a href=\"#r-16\">16</a>]. Giving streptokinase can also trigger immunogenicity which can confine numerous other treatments for a patient [<a href=\"#r-17\">17</a>]. Besides, selective third-generation thrombolytic agents including monoplane, tenecteplase, reteplase, etc. are contributing with notable angiographic potency in patients with acute myocardial infarction in spite of showing parallel mortality rates with those few agents that have been evaluated in large-scale trials [<a href=\"#r-14\">14</a>, <a href=\"#r-18\">18</a>]. Thus, persistent research is ongoing to offer advanced thrombolytic agents with utmost coronary arterial thrombolysis with nominal bleeding [<a href=\"#r-19\">19</a>] as currently almost all available synthetically derived thrombolytic agents are possessing notorious drawbacks [<a href=\"#r-20\">20</a>]. In accordance with a research, almost 30 % of all manufactured pharmaceuticals are plant-derived which are less toxic and safe from adverse effects than synthetically derived ones [<a href=\"#r-21\">21</a>]. Herbs and natural food sources along with their supplements having anticoagulant and antiplatelet activity can provide protection from detrimental coronary events and strokes [<a href=\"#r-17\">17</a>]. This incidence accredits extensive researches to ascertain new natural plant-derived sources of thrombolytic agents, as well as antimitotic agents [<a href=\"#r-22\">22</a>].<br />\r\nWith the ability to make humans and cattle vulnerable to fungal and bacterial infections, helminths have become the most familiar infectious agents in developing countries and less developed countries which mainly impose a great threat to public health especially to millions of school-going children [<a href=\"#r-23\">23</a>]. Synthetic anthelmintics with several drawbacks make the desired competence apocryphal including resistance [<a href=\"#r-24\">24</a>]. Previous research conducted on the efficacy of anthelmintics reflects that albendazole, levamisole, tetramisole, and ivermectin all can be compromised with resistance issues [<a href=\"#r-25\">25</a>]. One significant approach to embellish cheaper and efficient anthelmintics is to focus on herbal remedies. Thus, evaluation of anthelmintics derived from natural sources is becoming popular day by day in the treatment of parasite infections [<a href=\"#r-4\">4</a>]. Treatments with synthetic regional formulations have several adverse effects and are not available to low income people due to higher drug costs. Demand of medicinal plants is increasing gradually in the search for new medicinal drug for the treatment of several disorders around the world [<a href=\"#r-26\">26</a>].<br />\r\n<em>Tetrastigma leucostaphylum </em>is a woody climber belonging to the family of Vitaceae. Leaves of <em>T. leucostaphylum</em> (in wild) can coexist as simple, defoliate, and trifoliate patterns in the same plant [<a href=\"#r-27\">27</a>]. The genus <em>Tetrastigma</em> is comprised of 100 species [<a href=\"#r-28\">28</a>] which are seemingly found in the tropical or subtropical areas of Asian, and Australian undistributed rainforest [<a href=\"#r-29\">29</a>]. Parasitic plants of the Raffesiaceae family, producing the largest flower in the world utilize <em>Tetrastigma </em>as sole host [<a href=\"#r-30\">30</a>]. In Malaysia, Indonesia, and Vietnam leaf poultice or extracts of <em>Tetrastigma</em> plants are applied externally or internally to combat headaches and fever [<a href=\"#r-31\">31</a>]. Few species of <em>Tetrastigma</em> are well-known to treat sore throat, asthma, pneumonia, rheumatism, diarrhea, hepatitis, febrile convulsion, menstrual disorders, scrofula, immune system disorders, and cancer [<a href=\"#r-28\">28</a>]. The local name of <em>T. leucostaphylum</em> is Horina lata and has folkloric importance in tribal vicinity of Bangladesh (Chakma, Marma, and Tripura). Previously conducted research suggests that extract of <em>T. leucostaphylum</em> is rich in alkaloids, cardiac glycosides, carbohydrates, diterpenes, reducing sugars, phytosterols, saponin, fixed oils and fats and can be used as anesthetics and CNS stimulants due to high alkaloid content [<a href=\"#r-32\">32</a>]. Traditional practice of this plant also includes preventing fecundity, blood feeding, hatching of eggs along with diarrhea and dysentery management [<a href=\"#r-33\">33, 34</a>]. Pharmacological investigations of different plant extracts reveals the medicinal properties of many botanicals, but yet a lot of medicinal plants remain out of investigations. Hence isolation and characterization of healing compounds from these medicinal plants are needed [<a href=\"#r-35\">35</a>]. The research on the medicinal plants should be extended with the identification of the active principles in the plants. Scientific exploration of the remedies is composed of standardization and quality control of the products which is an evaluation procedure to get approval to be used in primary health care by ensuring their safety. Such research activities can also lead to the development of new drugs as in the future [<a href=\"#r-36\">36</a>]. As the best knowledge there is no investigations have been performed regarding the pain, thrombus, and anthelmintic management of this plant extract, this study has been intended to perform the analgesic, anti-inflammatory, thrombolysis and anthelmintic investigation of methanol extract of <em>T. leucostaphylum</em>.</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Collection and identification of the plant</strong><br />\r\nLeaves of <em>T. leucostaphylum</em> were collected from Hathazari hill, Chittagong hill region, Chittagong, Bangladesh in September 2018. Superior steps were taken to avoid contamination and healthy, fresh products have been reserved. The plant specimen was identified by Sajib Rudra, taxonomist, Bangladesh, and listed for ease of future reference (accession no: CTGUH SR7912) and stored in the Herbarium in the Chittagong University.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Extraction process</strong><br />\r\nAround 500 g of the dried powdered plant materials were taken in separate clean glass bottles and soaked in 2.5 liters of methanol. The container with its materials was fixed by aluminum foil with a container lid and kept for a period of 14 days at 23 ± 2⁰ C going with frequent shaking and mixing. The entire mixture was then filtered by cotton followed by the number 1 Whatman filter paper (Bibby RE200, Sterilin Ltd., UK) and the filtrates were condensed on a water bath at 40⁰ C, in this manner solvent can be evaporated [<a href=\"#r-37\">37</a>]. Finally, 12 g of the crude methanol extracts w</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Drugs and chemicals</strong><br />\r\nAnalytical grade chemicals and drugs have been used in this research and were purchased from BDH (UK) Laboratory Supplies and Sigma (St. Louis, MO, USA), Sanofi Bangladesh Ltd (Tongi, Bangladesh), Square Pharmaceuticals Ltd (Dhaka, Bangladesh) and Beximco Pharmaceuticals Ltd (Dhaka, Bangladesh).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Experimental animals</strong><br />\r\nStandard guidelines for the use and treatment of laboratory animals are extended to the handling and care of animals for the study [<a href=\"#r-38\">38</a>]. Swiss albino mice (weighing about 25–30 g), had been purchased from Jahangir Nagar University, Savar, Bangladesh. The animals received standard laboratory diet and water (<em>ad libitum</em>) and followed sufficient ventilation within the room following the normal day-night cycle. All the experiments were carried out in a separate and quiet state. For 10 days before the test, the animals have been adapted to the research laboratory environs. The designed protocol was approved by the P & D (Pharm P&D – 09/18-193) committee of Department of Pharmacy, International Islamic University Chittagong, Chittagong – 4318, Bangladesh.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong><em>In vivo</em></strong><strong> investigations</strong><br />\r\n<em>Acute toxicity test</em><br />\r\nPreviously described method approved by Organization for Economic Cooperation and Development (OECD) [<a href=\"#r-33\">33</a>, <a href=\"#r-39\">39</a>] with slight modifications has been used to investigate oral acute toxicity of the plant extract. Group of 10 mice received oral doses of 1000 mg/kg, 2000 mg/kg and 3000 mg/kg of METL while the control group received only the vehicle (water). For 48 hours, the groups were monitored. Every day the animals were weighed, and changes were noted in their typical behavior and lack of any signs of harmfulness.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Analgesic and anti-inflammatory investigations</strong><br />\r\n<em>Acetic acid-induced writhing test</em><br />\r\nThe peripheral nociceptive effect of the mice sample was determined by following the acetic acid-induced writhing test. This research has been performed by following the previously established method mentioned by Du, Junrong, et al. [<a href=\"#r-40\">40</a>]. Mice were divided into 4 groups and each group contained 5 mice. The first group was treated as a control group. They were administered orally with vehicle (1% Tween 80 in distilled water). In the second group, mice were treated with diclofenac-sodium (10 mg/kg, b.w; p.o). Third and fourth groups were administered orally with METL 200 and 400 (mg/kg, b.w; p.o) respectively 30 minutes before intraperitoneal injection of 0.6 % acetic acid solution at a dose of 10 mL/kg body weight. After 5 minutes, each group of mice observed for 30 minutes to count the number of writhing responses.<br />\r\nInhibition (%) =(Number of writhings by control-Number of writhings by test sample)/(Number of writhings by control)×100</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>Formalin induced paw licking test</em><br />\r\nTo evaluate the anti-inflammatory response of METL formalin mediated inflammatory studies were followed [<a href=\"#r-41\">41</a>]. At first, control (1% Tween 80), standard (ibuprofen 10 mg/kg), and test samples (METL 200 mg/kg, and 400 mg/kg) were administered orally. One hour later, 0.2 µL of 2.5% formalin was injected into the sub-plantar of the right-hand paw. Following formalin injection, the mice were immediately kept in a 40 cm<sup>3</sup> diameter jar, and licking time was observed. Licking of the paw was reported and observations expressed as a total licking time in the early stage (0-5 minutes) and the late stage (15-30 minutes) which was representing as neurogenic and inflammatory pain responses respectively.<br />\r\nInhibition (%) =(Number of lickings by control-Number of lickings by test sample)/(Number of lickings by control)× 100</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong><em>In vitro</em></strong><strong> investigations</strong><br />\r\n<strong><em>Thrombolytic assay</em></strong><br />\r\n<em>Streptokinase (SK) solution preparation</em><br />\r\n5 mL of sterile distill waters have been properly mixed into the commercially produced lyophilic SK vial (15,00,000 I.U) (PolaminWerk GmbH, Herdecque). This suspension was used as a stock solution for the <em>in vitro</em> thrombolysis investigation and finally, 30,000 I.U solutions were used from the stock solution.</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>Specimen for thrombolytic test</em><br />\r\nThe procedure referred to by Emon, et al. [<a href=\"#r-42\">42</a>] was followed in order to perform this analysis. Volunteers’ venous blood (500 μL / tube) was held in the preselected sterile Eppendorf tube, and incubating at a temperature of 37 ° C for 45 minutes. The serum was completely separated after the development of the coagulation. Each tube reweighed to verify the weight of the clot. After removing the clot, 100 μL of METL has been correctly applied to each Eppendorf. Finally, each tube was incubated again for the 90 minutes at 37 °C coagulation. After incubation, the fluid found was detached and tubes were again weighed to detect the change in weight after clot disruption. The weight difference before and after clot lysis was considered to the final percentage of clot lysis. Water and streptokinase have been used as the positive and the negative and positive controls respectively. The following formula has been followed to determine the percent of clot lysis.<br />\r\n% clot lysis =(Weight of the lysis clot)/(Weight of clot before lysis)×100.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong><em>Anthelmintic assay</em></strong><br />\r\nPreviously described method [<a href=\"#r-43\">43</a>] with slight modification, the anthelmintic research has been implemented. Nearly 6-8 worms (<em>Tubifex tubifex</em>) were taken in each of six Petri dishes, and METL at different concentrations (10 mg/mL, 5 mg/mL, 2.5 mg/mL) were added in 3 Petri dishes and standard levamisole at different concentrations (1 mg/mL, 0.8 mg/mL, 0.5 mg/mL) were added in rest 3 Petri dishes. Then the beginning of the paralysis and the time of the worms’ death were observed carefully. In two distinct parameters, ‘time for paralysis’ and ‘time for death’ of the worms were determined to distinguish the anthelmintic effect of the sample. Paralysis time was recorded if no movement could be detected. Upon confirmation, time taken to kill worms was noted if the worms did not react either when shaken vigorously or when plunged into the slightly warm water.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Statistical analysis</strong><br />\r\nStatistical analysis was interpreted as mean ± standard error (SEM) and the data were analyzed using GraphPad Prism 5.2 (GraphPad Software, Inc., La Jolla, CA, United States). Statistical significance was determined by a one-way variance analysis (ANOVA) followed by Dunnett’s test, where *<em>P</em> < 0.5, **<em>P</em> < 0.01, and ***<em>P</em> < 0.001 was considered statistically significant.</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><strong>Effect of extracts on oral acute toxicity test</strong><br />\r\nDuring the observations, there was no lethality, no behavioral change (sedation, excitability) or no allergies reaction was appeared after the oral administration of METL.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Effect of the extracts on acetic acid-induced writhing test</strong><br />\r\nPeripheral analgesic activity determination by the acetic acid-induced writhing study was measured on the basis of the average number of abdominal constrictions shown during the writhing test by extending a hind paw to animals (mice). Inhibition was observed for 20 min in the writhing of the test extract. In contrast to the control group, a dose of 200 mg/kg inhibited 8.28% of writhes. Dose of 400 mg/kg inhibited 31.31 % (<em>P</em> < 0.05) of writhes while the standard drug diclofenac-sodium exhibited 66.58 % (<em>P</em> < 0.001) of inhibition. The overall results of the control and test samples have been narrated in <a href=\"#Table-1\">Table 1</a>. Comparing with standard drug diclofenac-sodium, the therapeutic activity of METL was less than the standard drug diclofenac sodium.</p>\r\n\r\n<div id=\"Table-1\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1590260727-table1/\">Table-1</a><strong>Table 1.</strong> Effect of test samples on the analgesic study of the METL on the acetic acid-induced writhing test in mice.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Effect of extracts on formalin-induced paw licking test</strong><br />\r\nThe formalin test exhibited moderate anti-inflammatory activity. When METL was administered orally at 200 and 400 mg/kg according to body weight, the licking time in both the early and late phases was decreased in a dose-dependent manner. METL at 200 mg/kg caused significant inhibition of licking in both early phase (28.34 %) (<em>P</em> < 0.01) and late phase (33.72 %) (<em>P</em> < 0.01). METL at 400 mg/kg also caused significant inhibition of licking in early phase (39.63%) (<em>P</em> < 0.01) and late phase (48.73%) (<em>P</em> < 0.001). Standard drug diclofenac-sodium inhibited paw licking 53.39 % (<em>P</em> < 0.001) in early phase and 54.67% (<em>P</em> < 0.001) in late phase when compared with control (1% tween 80) group animals. The summary of the findings has been enumerated in <a href=\"#figure1\">Figure 1</a>.</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"314\" src=\"/media/article_images/2024/09/14/178-1590260727-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1. </strong>Effect of test samples on the anti-inflammatory investigation of the METL on the formalin-induced licking test. The values were presented as mean ± SEM; One-way analysis of variance (ANOVA) was followed by Dunnett’s test. *<em>P</em> < 0.05, **<em>P </em>< 0.01 and ***<em>P</em> < 0.001, where # was designated as control. METL = methanol extract of <em>Tetrastigma leucostaphylum</em> leaves, TWN 80 = 1% Tween 80, and IBU = Ibuprofen.</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Effect of the extracts on thrombolytic activity</strong><br />\r\nIn the <em>in vitro</em> thrombolytic assay, the extract exerted 26.08 % (<em>P</em> < 0.001) lysis of the blood clot while standard streptokinase was obtained 56.62 % (<em>P</em> < 0.001) blood clot lysis. The summary of the findings has been enumerated in <a href=\"#figure2\">Figure 2</a>.</p>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"357\" src=\"/media/article_images/2024/09/14/178-1590260727-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2.</strong> Clot lysis effects by Saline water, Streptokinase, and METL. The values were presented as mean ± SEM (n = 10); One-way analysis of variance (ANOVA) was followed by Dunnett’s test. ***<em>P</em> < 0.001 was considered as significant compared with the control, where # is designated as control. METL = methanol extract of <em>Tetrastigma leucostaphylum</em> leaves, SW = saline water, and STK = streptokinase.</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Effect of extracts on </strong><strong>anthelmintic activity</strong><br />\r\nA methanol extract of <em>T. leucostaphylum </em>leaves was used to determine the anthelmintic activity and the extract showed the effect in a dose-dependent manner. METL took 9.3 ± 1.16 minutes for paralysis and 32.45 ± 2.35 minutes for death at the maximum concentration (10 mg/mL) while standard levamisole took 4.5 ± 1.38 minutes for paralysis and 8.55 ± 1.33 minutes for death at a concentration of 1 mg/mL (<a href=\"#figure3\">Figure 3</a>).</p>\r\n\r\n<div id=\"figure3\">\r\n<figure class=\"image\"><img alt=\"\" height=\"330\" src=\"/media/article_images/2024/09/14/178-1590260727-Figure3.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 3. </strong>Effect of various test doses of the METL and LMS on nematodes (n = 6) in the anthelmintic study. Values were presented as mean ± SEM. METL = methanol extract of <em>Tetrastigma leucostaphylum</em> leaves, and LMS = levamisole.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>The study was performed to investigate the analgesic, anti-inflammatory, thrombolytic and anthelmintic significance of methanol extract of <em>T. leucostaphylum</em> leaves. The experiments showed negligible anti-nociception activity after the oral administration of METL and not significantly decreased abdominal writhing in mice. Acetic acid-induced abdominal restriction response is an established procedure in peripheral analgesic agent research. Prostaglandin receptors are thought to mediate this response [<a href=\"#r-44\">44</a>]. METL has generated less anti-nociceptive behavior and therefore suggests that the prostaglandin receptor secretion could not be attenuated strongly by the components of METL. Throughout the scientific studies, METL has shown dose-dependent activity in anti-inflammatory research. For the justification of pain and analgesia the formalin-induced pain approach is quite worthwhile [<a href=\"#r-45\">45</a>]. The formalin-induced pain experiment includes two stages of painful exposure [<a href=\"#r-46\">46</a>]. The first stage is the early (neurogenic pain) stage which usually results from peripheral stimulation through C-fibre [<a href=\"#r-47\">47</a>]. This stage begins immediately after the injection of the formalin solution and takes 5-10 minutes. The 2nd (late) stage (inflammatory pain) is intensified by inflammation of local tissues and physiological alteration of the dorsal horn of the spinal cord which begins about 20 minutes after the formalin injection. These process has been inhibited by opioid and anti-inflammatory medicines [<a href=\"#r-41\">41</a>]. Therefore, central analgesics such as morphine often block early and late phases while medications like adrenaline (dexamethasone), or NSAIDs (aspirin), often inhibit pain in late-phase [<a href=\"#r-48\">48</a>]. In formalin study, it was assumed that METL substantially (<em>P</em> < 0.01 and <em>P</em> < 0.001) attenuated the hyperalgesia in both phases which has been mediated throughout the formalin injection. Disruption of hemostasis can cause a blood clot (thrombus) in the circulatory system and results in vascular blockage followed by detrimental consequences in thrombolytic diseases including acute myocardial or cerebral infarction which may lead to death [<a href=\"#r-1\">1</a>].<br />\r\nThrombolysis is the sum of several detrimental cascades related to arterial diseases connected with acute coronary disorders such as pulmonary emboli, deep vein thrombosis, strokes, heart attacks, and venous thromboembolic complications which can result in abrupt morbidity and mortality [<a href=\"#r-12\">12</a>]. In the physiological system, vascular barricade results from thrombosis, and recuperating events can lead to fatal consequences including cerebral or myocardial infarction followed by death [<a href=\"#r-20\">20</a>]. The hypothesis of cascade and waterfall [<a href=\"#r-49\">49, 50</a>] shows that the coagulation process takes place in three stages: the formation of the thrombin, activation of prothrombin, and formation of fibrin. The results of this study have shown that METL has enabled lysis to the extrinsic coagulation which may act as the driving force of antithrombotic and thrombolytic action. Furthermore, the low dose intake of crude extract has no or fewer side effects and the body’s coagulation system will not be hampered. For long-term thrombosis prevention, low dose METL is therefore recommended.<br />\r\nIn the anthelminthic study, METL increased the paralysis and death of <em>Tubifex tubifex</em> within a short time in a dose-dependent manner. Helminths, worm-like organisms reside inside the living host and assemble nourishment from it which results in disruption in hosts nutrient absorption mechanism. Through skin or gastrointestinal tract juvenile forms of the parasites can invade human beings and go under maturation to form adult worms by arresting nutrition [<a href=\"#r-51\">51</a>]. Anthelmintics, a group of antiparasitic drugs perform locally to expel worms from the gastrointestinal tract [<a href=\"#r-52\">52</a>] or systematically to restrict helminths from doing any noteworthy damage to host [<a href=\"#r-53\">53</a>]. METL’s anthelmintic behavior may be exhibited due to the involvement of the phytoconstituents of the plant extract. Phytoconstituents can interfere with the generation of energy by de-aligning oxidative phosphorylation, or due to the interfere of phytoconstitents with the glycoproteins of the cell surface.</p>"
},
{
"section_number": 5,
"section_title": "CONCLUSIONS",
"body": "<p>The study revealed significant anti-inflammatory, thrombolytic, and anthelmintic potentiality of methanol extract of<em> T. leucostaphylum</em> leaves and can be considered as a prominent candidate to be a lead compound for drug discovery and drug designing. Chemical dynamics of the plant extract has been presumed to trigger the biological activity of the plant extract. Finally, further investigations following chemical profiling are recommended to confirm the biological activity of the plant extract.</p>"
},
{
"section_number": 6,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>The authors are thankful to the Department of Pharmacy, International Islamic University Chittagong, Chittagong – 4318, Bangladesh and “The Article (Road to Drug Discovery)” for the laboratory, and other aids required for this research. The authors also would like to thank Mrs. Sultana Razia Shorna (Chittagong college, Chittagong – 4203, Bangladesh) for her heartfelt cooperation. No particular grant was received from public, private or non-profit funding agencies for this research.</p>"
},
{
"section_number": 7,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>SR, SUS, and NUE conceptualized, designed, and prepared the research protocols. SUS and NUE performed the laboratory experiments. MMRK, AMSC, ANH, and MS curated the data. SAS, SR, SA and NUE interpreted the data. SA, NUE and SUS drafted the manuscript. SMT and MAS supervised while MNI monitored the research. Finally, all the authors revise the final draft and agreed to publish the manuscript as an original research article.</p>"
},
{
"section_number": 8,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/09/14/178-1590260727-Figure1.jpg",
"caption": "Figure 1. Effect of test samples on the anti-inflammatory investigation of the METL on the formalin-induced licking test. The values were presented as mean ± SEM; One-way analysis of variance (ANOVA) was followed by Dunnett’s test. *P < 0.05, **P < 0.01 and ***P < 0.001, where # was designated as control. METL = methanol extract of Tetrastigma leucostaphylum leaves, TWN 80 = 1% Tween 80, and IBU = Ibuprofen.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/09/14/178-1590260727-Figure2.jpg",
"caption": "Figure 2. Clot lysis effects by Saline water, Streptokinase, and METL. The values were presented as mean ± SEM (n = 10); One-way analysis of variance (ANOVA) was followed by Dunnett’s test. ***P < 0.001 was considered as significant compared with the control, where # is designated as control. METL = methanol extract of Tetrastigma leucostaphylum leaves, SW = saline water, and STK = streptokinase.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/09/14/178-1590260727-Figure3.jpg",
"caption": "Figure 3. Effect of various test doses of the METL and LMS on nematodes (n = 6) in the anthelmintic study. Values were presented as mean ± SEM. METL = methanol extract of Tetrastigma leucostaphylum leaves, and LMS = levamisole.",
"featured": false
}
],
"authors": [
{
"id": 876,
"affiliation": [
{
"affiliation": "Department of Botany, Faculty of Biological Science, University of Chittagong, Chittagong - 4331, Bangladesh"
}
],
"first_name": "Sajib",
"family_name": "Rudra",
"email": null,
"author_order": 1,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 877,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Md. Solaman Uddin",
"family_name": "Sawon",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 878,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Nazim Uddin",
"family_name": "Emon",
"email": "nazim7emon@gmail.com",
"author_order": 3,
"ORCID": "https://orcid.org/0000-0001-7567-4796",
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Nazim Uddin Emon, Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh, E-mail: nazim7emon@gmail.com",
"article": 200
},
{
"id": 879,
"affiliation": [
{
"affiliation": "Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Dhaka, Dhaka-1000, Bangladesh."
}
],
"first_name": "Safaet",
"family_name": "Alam",
"email": null,
"author_order": 4,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 880,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Syed Mohammed",
"family_name": "Tareq",
"email": null,
"author_order": 5,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 881,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Mohammad Nazmul",
"family_name": "Islam",
"email": null,
"author_order": 6,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 882,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Mir Md. Rokib",
"family_name": "Uddin",
"email": null,
"author_order": 7,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 883,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Abdullah Md Sazid",
"family_name": "Chowdhury",
"email": null,
"author_order": 8,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 884,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Abu Nazer",
"family_name": "Hasbe",
"email": null,
"author_order": 9,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 885,
"affiliation": [
{
"affiliation": "Department of Botany, Faculty of Biological Science, University of Chittagong, Chittagong - 4331, Bangladesh"
}
],
"first_name": "Mohammad",
"family_name": "Shakil",
"email": null,
"author_order": 10,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 886,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Shahenur Alam",
"family_name": "Sakib",
"email": null,
"author_order": 11,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
},
{
"id": 887,
"affiliation": [
{
"affiliation": "Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong - 4318, Bangladesh"
}
],
"first_name": "Mohammed Aktar",
"family_name": "Sayeed",
"email": null,
"author_order": 12,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 200
}
],
"views": 603,
"downloads": 70,
"references": [
{
"id": 6721,
"serial_number": 1,
"pmc": null,
"reference": "Viana F. Nociceptors: thermal allodynia and thermal pain. Handbook of clinical neurology: Elsevier; 2018. p. 103-19.",
"DOI": null,
"article": 200
},
{
"id": 6722,
"serial_number": 2,
"pmc": null,
"reference": "Ferrero‐Miliani L, Nielsen O, Andersen P, Girardin S. Chronic inflammation: importance of NOD2 and NALP3 in interleukin‐1β generation. Clinical & Experimental Immunology. 2007;147:227-35.",
"DOI": null,
"article": 200
},
{
"id": 6723,
"serial_number": 3,
"pmc": null,
"reference": "Calixto JB, Beirith A, Ferreira J, Santos AR, Filho VC, Yunes RA. Naturally occurring antinociceptive substances from plants. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2000;14:401-18.",
"DOI": null,
"article": 200
},
{
"id": 6724,
"serial_number": 4,
"pmc": null,
"reference": "Farnsworth N. Screening Plants for New Medicines. Biodiversity. Part II. Washington: National Academy Press; 1989.",
"DOI": null,
"article": 200
},
{
"id": 6725,
"serial_number": 5,
"pmc": null,
"reference": "Elekwa I. Preliminary phytochemical screening and gas chromatographic FID evaluation of Garcinia kola seed extracts. Journal of Pharmacognosy and Phytochemistry. 2014;2.",
"DOI": null,
"article": 200
},
{
"id": 6726,
"serial_number": 6,
"pmc": null,
"reference": "Musa Y, Haruna A, Ilyas M, Yaro A, Ahmadu A, Usman H. Phytochemical, analgesic and anti-inflammatory effects of the ethylacetate extract of the leaves of Pseudocedrella kotschyii. African Journal of Traditional, Complementary and Alternative Medicines. 2008;5:92-6.",
"DOI": null,
"article": 200
},
{
"id": 6727,
"serial_number": 7,
"pmc": null,
"reference": "de Melo MS, Quintans JdSS, Araújo AAdS, Duarte MC, Bonjardim LR, Nogueira PCdL, et al. A systematic review for anti-inflammatory property of clusiaceae family: a preclinical approach. J Evid Based Integr Med. 2014;2014.",
"DOI": null,
"article": 200
},
{
"id": 6728,
"serial_number": 8,
"pmc": null,
"reference": "Yasmen N, Aziz M, Tajmim A, Akter M, Hazra AK, Rahman S. Analgesic and anti-inflammatory activities of diethyl ether and n-hexane extract of polyalthia suberosa leaves. J Evid Based Integr Med. 2018;2018.",
"DOI": null,
"article": 200
},
{
"id": 6729,
"serial_number": 9,
"pmc": null,
"reference": "Ingrasciotta Y, Sultana J, Giorgianni F, Fontana A, Santangelo A, Tari DU, et al. Association of individual non-steroidal anti-inflammatory drugs and chronic kidney disease: a population-based case control study. PLoS One. 2015;10.",
"DOI": null,
"article": 200
},
{
"id": 6730,
"serial_number": 10,
"pmc": null,
"reference": "Shih S-C, Chang C-W. Nonsteroidal anti-inflammatory drug-related gastrointestinal bleeding in the elderly. International Journal of Gerontology. 2007;1:40-5.",
"DOI": null,
"article": 200
},
{
"id": 6731,
"serial_number": 11,
"pmc": null,
"reference": "Howes M-JR. Phytochemicals as anti-inflammatory nutraceuticals and phytopharmaceuticals. Immunity and Inflammation in Health and Disease: Elsevier; 2018. p. 363-88.",
"DOI": null,
"article": 200
},
{
"id": 6732,
"serial_number": 12,
"pmc": null,
"reference": "Mahmud S, Akhter S, Rahman M, Aklima J, Akhter S, Merry SR, et al. Antithrombotic effects of five organic extracts of Bangladeshi plants in vitro and mechanisms in in silico models. J Evid Based Integr Med. 2015;2015.",
"DOI": null,
"article": 200
},
{
"id": 6733,
"serial_number": 13,
"pmc": null,
"reference": "Fathima SN, Ahmad SV, Kumar BR. Evaluation of in vitro thrombolytic activity of ethanolic extract of Curcuma caesia rhizomes. International Journal of Pharma Research & Review. 2015;4:50-4.",
"DOI": null,
"article": 200
},
{
"id": 6734,
"serial_number": 14,
"pmc": null,
"reference": "Sayeed MA, Kabir H, Rashid MMU, Bhuiyan MFA, Rashid MA. Thrombolytic activity of methanolic extracts of Desmodium paniculatum (L.) and Sarcochlamys pulcherrima (Roxb.). Bangladesh Pharmaceutical Journal. 2014;17:67-9.",
"DOI": null,
"article": 200
},
{
"id": 6735,
"serial_number": 15,
"pmc": null,
"reference": "Dibyajyoti S, Swati P. Phytochemical Screening and Thrombolytic Activity of Chloroform Extract of Urena Sinuata (L.). Journal of Fundamental and Applied Sciences. 2014;6:208-19.",
"DOI": null,
"article": 200
},
{
"id": 6736,
"serial_number": 16,
"pmc": null,
"reference": "Mannan A, Kawser MJ, Ahmed AA, Islam NN, Alam SM, Khan MAE, et al. Assessment of antibacterial, thrombolytic and cytotoxic potential of Cassia alata seed oil. Journal of Applied Pharmaceutical Science. 2011;1:56.",
"DOI": null,
"article": 200
},
{
"id": 6737,
"serial_number": 17,
"pmc": null,
"reference": "Furie B, Furie BC. Mechanisms of thrombus formation. New England Journal of Medicine. 2008;359:938-49.",
"DOI": null,
"article": 200
},
{
"id": 6738,
"serial_number": 18,
"pmc": null,
"reference": "Emon NU, Kaiser M, Islam M, Kabir MFI, Jamir M, Uddin MAJ, et al. Anxiolytic and thrombolytic investigation of methanol extract of Piper nigrum L. fruits and Sesamum indicum L. seeds.",
"DOI": null,
"article": 200
},
{
"id": 6739,
"serial_number": 19,
"pmc": null,
"reference": "Collen D. Coronary thrombolysis: streptokinase or recombinant tissue-type plasminogen activator? Annals of internal medicine. 2020.",
"DOI": null,
"article": 200
},
{
"id": 6740,
"serial_number": 20,
"pmc": null,
"reference": "Nicolini FA, Nichols WW, Mehta JL, Saldeen TG, Schofield R, Ross M, et al. Sustained reflow in dogs with coronary thrombosis with K2P, a novel mutant of tissue-plasminogen activator. Journal of the American College of Cardiology. 1992;20:228-35.",
"DOI": null,
"article": 200
},
{
"id": 6741,
"serial_number": 21,
"pmc": null,
"reference": "Annapurna A, Mahalakshmi DK, Krishna KM. Antidiabetic activity of a poly herbal preparation (tincture of panchparna) in normal and diabetic rats. 2001.",
"DOI": null,
"article": 200
},
{
"id": 6742,
"serial_number": 22,
"pmc": null,
"reference": "Wagnor H, Hikino H, Farnsworth R, London N. The economic significance of plants and their constituents as drug. Economic and medicinal plant research. 1989;3:1-17.",
"DOI": null,
"article": 200
},
{
"id": 6743,
"serial_number": 23,
"pmc": null,
"reference": "Silva Soares SC, de Lima GC, Carlos Laurentiz A, Féboli A, dos Anjos LA, de Paula Carlis MS, et al. In vitro anthelmintic activity of grape pomace extract against gastrointestinal nematodes of naturally infected sheep. International journal of veterinary science and medicine. 2018;6:243-7.",
"DOI": null,
"article": 200
},
{
"id": 6744,
"serial_number": 24,
"pmc": null,
"reference": "Zenebe S, Feyera T, Assefa S. In vitro anthelmintic activity of crude extracts of aerial parts of Cissus quadrangularis L. and leaves of Schinus molle L. against Haemonchus contortus. BioMed research international. 2017;2017.",
"DOI": null,
"article": 200
},
{
"id": 6745,
"serial_number": 25,
"pmc": null,
"reference": "Wakayo B, Dewo T. Anthelmintic resistance of gastrointestinal parasites in small ruminants: a review of the case of Ethiopia. J Veterinar Sci Technol S. 2015;10.",
"DOI": null,
"article": 200
},
{
"id": 6746,
"serial_number": 26,
"pmc": null,
"reference": "Herrera-Ruiz M, García-Beltrán Y, Mora S, Díaz-Véliz G, Viana GS, Tortoriello J, et al. Antidepressant and anxiolytic effects of hydroalcoholic extract from Salvia elegans. J Ethnopharmaco. 2006;107:53-8.",
"DOI": null,
"article": 200
},
{
"id": 6747,
"serial_number": 27,
"pmc": null,
"reference": "Krishna T, Krishna T, Chithra N, Deepa P, Darsana U, Sreelekha K, et al. Acaricidal activity of petroleum ether extract of leaves of Tetrastigma leucostaphylum (Dennst.) Alston against Rhipicephalus (Boophilus) annulatus. The Scientific World Journal. 2014;2014.",
"DOI": null,
"article": 200
},
{
"id": 6748,
"serial_number": 28,
"pmc": null,
"reference": "Adarsh Krishna T, Ajeesh Krishna T, Sanyo Raj V, Juliet S, Nair S, Ravindran R, et al. Evaluation of phytochemical constituents and proximate contents of the ethanolic leaf extract of Tetrastigmaleucostaphylum (Dennst.) Alstone (Vitaceae) found in Western Ghats of Kerala, India. Research Journal of Pharmaceutical Sciences ISSN. 2013;2319:555X.",
"DOI": null,
"article": 200
},
{
"id": 6749,
"serial_number": 29,
"pmc": null,
"reference": "Hossain MA, Shah MD, Gnanaraj C, Iqbal M. In vitro total phenolics, flavonoids contents and antioxidant activity of essential oil, various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah. Asian Pac J Trop Med. 2011;4:717-21.",
"DOI": null,
"article": 200
},
{
"id": 6750,
"serial_number": 30,
"pmc": null,
"reference": "Barcelona JF, Pelser PB, Cajano MO. Rafflesia banahaw (Rafflesiaceae), a new species from Luzon, Philippines. Blumea-Biodiversity, Evolution and Biogeography of Plants. 2007;52:345-50.",
"DOI": null,
"article": 200
},
{
"id": 6751,
"serial_number": 31,
"pmc": null,
"reference": "Dao PTA, Le Quan T, Mai NTT. Some compounds from stem of tetrastigma erubescens planch.(vitateae). Journal of Engineering Technology and Education. 2013;9:54-7.",
"DOI": null,
"article": 200
},
{
"id": 6752,
"serial_number": 32,
"pmc": null,
"reference": "Madziga H, Sanni S, Sandabe U. Phytochemical and elemental analysis of Acalypha wilkesiana leaf. Journal of American Science. 2010;6:510-4.",
"DOI": null,
"article": 200
},
{
"id": 6753,
"serial_number": 33,
"pmc": null,
"reference": "Van den Heuvel M. A New Approach to the Classification of Substances and Preparations on the Basis of their Acute Toxicity: A Report by the British Toxicology Society* Working Party on Toxicity. Human Toxicology. 1984;3:85-92.",
"DOI": null,
"article": 200
},
{
"id": 6754,
"serial_number": 34,
"pmc": null,
"reference": "Ashraf MA, Khatun A, Sharmin T, Mobin F, Tanu AR, Morshed T, et al. MPDB 1.0: a medicinal plant database of Bangladesh. Bioinformation. 2014;10:384.",
"DOI": null,
"article": 200
},
{
"id": 6755,
"serial_number": 35,
"pmc": null,
"reference": "Tasleem F, Azhar I, Ali SN, Perveen S, Mahmood ZA. Analgesic and anti-inflammatory activities of Piper nigrum L. Asian Pac J Trop Med. 2014;7:S461-S8.",
"DOI": null,
"article": 200
},
{
"id": 6756,
"serial_number": 36,
"pmc": null,
"reference": "Pattanayak P, Behera P, Das D, Panda SK. Ocimum sanctum Linn. A reservoir plant for therapeutic applications: An overview. Pharmacognosy reviews. 2010;4:95.",
"DOI": null,
"article": 200
},
{
"id": 6757,
"serial_number": 37,
"pmc": null,
"reference": "Alam S, Emon NU, Rashid MA, Arman M, Haque MR. Investigation of biological activities of Colocasia gigantea Hook. f. leaves and PASS prediction, in silico molecular docking with ADME/T analysis of its isolated bioactive compounds. BioRxiv. 2020.",
"DOI": null,
"article": 200
},
{
"id": 6758,
"serial_number": 38,
"pmc": null,
"reference": "National Research Council Committee for the Update of the Guide for the C, Use of Laboratory A. The National Academies Collection: Reports funded by National Institutes of Health. In: th, editor. Guide for the Care and Use of Laboratory Animals. Washington (DC): National Academies Press (US) National Academy of Sciences.; 2011.",
"DOI": null,
"article": 200
},
{
"id": 6759,
"serial_number": 39,
"pmc": null,
"reference": "Walum E. Acute oral toxicity. Environmental health perspectives. 1998;106:497-503.",
"DOI": null,
"article": 200
},
{
"id": 6760,
"serial_number": 40,
"pmc": null,
"reference": "Du J, Yu Y, Ke Y, Wang C, Zhu L, Qian ZM. Ligustilide attenuates pain behavior induced by acetic acid or formalin. J Ethnopharmaco. 2007;112:211-4.",
"DOI": null,
"article": 200
},
{
"id": 6761,
"serial_number": 41,
"pmc": null,
"reference": "Emon NU, Jahan I, Sayeed MA. Investigation of antinociceptive, anti-inflammatory and thrombolytic activity of Caesalpinia digyna (Rottl.) leaves by experimental and computational approaches. Advances in Traditional Medicine. 2020:1-9.",
"DOI": null,
"article": 200
},
{
"id": 6762,
"serial_number": 42,
"pmc": null,
"reference": "Emon NU, Jahan I, Rudra S, Aktar M, Islam N. In Vitro Comparative Cardioprotective Activity of Methanol Extract of Caesalpinia Digyna (Rottl.) Stems and Senna Sophera (L.) Roxb. Stems.",
"DOI": null,
"article": 200
},
{
"id": 6763,
"serial_number": 43,
"pmc": null,
"reference": "Tariq K, Chishti M, Ahmad F, Shawl A. Anthelmintic activity of extracts of Artemisia absinthium against ovine nematodes. Veterinary parasitology. 2009;160:83-8.",
"DOI": null,
"article": 200
},
{
"id": 6764,
"serial_number": 44,
"pmc": null,
"reference": "Ribeiro RA, Vale ML, Thomazzi SM, Paschoalato AB, Poole S, Ferreira SH, et al. Involvement of resident macrophages and mast cells in the writhing nociceptive response induced by zymosan and acetic acid in mice. Eur J Pharmacol. 2000;387:111-8.",
"DOI": null,
"article": 200
},
{
"id": 6765,
"serial_number": 45,
"pmc": null,
"reference": "Jimoh AO, Chika A, Umar MT, Adebisi I, Abdullahi N. Analgesic effects and anti-inflammatory properties of the crude methanolic extract of Schwenckia americana Linn (Solanaceae). J Ethnopharmaco. 2011;137:543-6.",
"DOI": null,
"article": 200
},
{
"id": 6766,
"serial_number": 46,
"pmc": null,
"reference": "Moniruzzaman M, Imam MZ. Evaluation of antinociceptive effect of methanolic extract of leaves of Crataeva nurvala Buch.-Ham. BMC Complement Altern Med. 2014;14:354.",
"DOI": null,
"article": 200
},
{
"id": 6767,
"serial_number": 47,
"pmc": null,
"reference": "Wooten M, Weng H-J, Hartke TV, Borzan J, Klein AH, Turnquist B, et al. Three functionally distinct classes of C-fibre nociceptors in primates. Nature communications. 2014;5:1-12.",
"DOI": null,
"article": 200
},
{
"id": 6768,
"serial_number": 48,
"pmc": null,
"reference": "Pelayo VdRT, Villalba BS, Guadarrama AAD, Reyes AM, Pernas MÁC. Thermal nociceptive tail-flick reflex development in Wistar male rats. Revista Médica de la Universidad Veracruzana. 2007;7:8-11.",
"DOI": null,
"article": 200
},
{
"id": 6769,
"serial_number": 49,
"pmc": null,
"reference": "Riddel Jr JP, Aouizerat BE, Miaskowski C, Lillicrap DP. Theories of blood coagulation. Journal of Pediatric Oncology Nursing. 2007;24:123-31.",
"DOI": null,
"article": 200
},
{
"id": 6770,
"serial_number": 50,
"pmc": null,
"reference": "Toussaint EF, Short AE. Transoceanic stepping–stones between cretaceous waterfalls? The enigmatic biogeography of pantropical Oocyclus cascade beetles. Molecular phylogenetics and evolution. 2018;127:416-28.",
"DOI": null,
"article": 200
},
{
"id": 6771,
"serial_number": 51,
"pmc": null,
"reference": "Joseph B, George J, Mohan J. Pharmacology and traditional uses of Mimosa pudica. International journal of pharmaceutical sciences and drug research. 2013;5:41-4.",
"DOI": null,
"article": 200
},
{
"id": 6772,
"serial_number": 52,
"pmc": null,
"reference": "McSorley HJ, Chayé MA, Smits HH. Worms: Pernicious parasites or allies against allergies? Parasite immunology. 2019;41:e12574.",
"DOI": null,
"article": 200
},
{
"id": 6773,
"serial_number": 53,
"pmc": null,
"reference": "El-Rahman MA, Abdel-Nabi IM, Omran MA, Mohamed MF. Cytotoxic effects of albendazole, an antiparasitic drug, on the liver of the rat: subchronic study. Egyptian journal of Biology. 1999;1:16-29.",
"DOI": null,
"article": 200
}
]
},
{
"id": 193,
"slug": "178-1588348425-genetic-fingerprinting-for-the-protection-of-local-rice-oryza-sativa-l-cultivars-of-bangladesh",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1588348425",
"recieved": "2020-03-07",
"revised": null,
"accepted": "2020-05-15",
"published": "2020-06-13",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/35/178-1588348425.pdf",
"title": "Genetic fingerprinting for the protection of local rice (Oryza sativa L.) cultivars of Bangladesh",
"abstract": "<p>Genetic fingerprinting of 110 rice cultivars of Bangladesh was completed with five polymorphic microsatellite DNA markers such as RM153, RM251, RM333, RM335 and RM475. The amplified DNA fragments are known as alleles from Polymerase Chain Reaction (PCR) reactions were separated on 2% agarose gel electrophoresis system, subsequently visualized by high performance ultraviolet transilluminator. In all, 99 distinctive alleles averaging 19.80 alleles/locus from the entire utilized microsatellite loci were counted. Several diversity indexes such as Polymorphism Information Content (PIC), heterozygosity, and cluster analysis were computed in this quantitative investigation. Superior genetic differentiation and inferior gene flow values among the cultivars were revealed from the recorded genetic diversity study of PIC, Effective allele, Shannon index (<em>I</em>), Hardy-Weinberg equilibrium (HWE), Nei’s gene diversity (<em>h</em>), along with genetic differentiation-F<sub>is</sub> and gene flow-N<sub>m</sub> analysis. A total, 5995 varietal pairs were achieved all the way through alternative combinations of 110 rice cultivars where their Nei’s genetic distance (<em>D</em>) was ranged from zero to 2.832. Nei’s genetic-base an Unweight Pair Group Method of Arithmetic Means (UPGMA) diagram was assembled which eventually separated all the cultivars from each other according to their genetic distance and similarity. Thus, the finding of this study will expose such strategies to distinct all the wild relatives, cultivars and commercial varieties of rice or any other crop species having various genetic levels to facilitate further improvement and protection in future.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 182-193.",
"academic_editor": "Dr. Mohammad Nazrul Islam, Sher-e-Bangla Agricultural University, Bangladesh.",
"cite_info": "Rahman MS, Sohag MKH, et al. Genetic fingerprinting for the protection of local rice (Oryza sativa L.) cultivars of Bangladesh. J Adv Biotechnol Exp Ther. 2020; 3(3): 182-193.",
"keywords": [
"Bangladesh",
"Plant variety protection",
"Oryza sativa L.",
"Genetic fingerprinting"
],
"DOI": "10.5455/jabet.2020.d124",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>Rice (<em>Oryza sativa </em>L.) having diploid chromosome number 2n=2x=24 belongs to the significant grass family <em>Gramineae</em> and subfamily <em>Oryzoidae</em> is considered one of the most cultivated annual cereals because almost 1/2 of the world’s human population consume rice every day as their predominant staple food [<a href=\"#r-1\">1</a>]. It is occupied in the central position other than rest of the agricultural component to contribute Bangladesh’s national economy [<a href=\"#r-2\">2</a>]. It is believed that Asian farmers have been maintaining the selection and domestication process of a wide range of rice cultivars since ancient [<a href=\"#r-3\">3</a>]. The Gene bank of Bangladesh Rice Research Institute has accumulated near about 8,500 rice germplasm from different ecological and indigenous sources of Bangladesh of which 8,044 genotypes have been registered so far [<a href=\"#r-4\">4</a>]. Great variations in rice genotypes have been noticed in Asia, more particularly in China as well as Indian subcontinent regarding morphological, biochemical and molecular aspects [<a href=\"#r-5\">5</a>],[<a href=\"#r-6\">6</a>]. Zhao [<a href=\"#r-7\">7</a>] has reported on the subject of the origin of rice which has been accomplished close to the northeastern part of India (Assam) and southwestern part of China (Yunan), both are truly recommended to the subtropical upland of Himalayas [<a href=\"#r-8\">8</a>]. <em>Oryza rufipogon </em>and <em>Oryza nivara </em>are the two parts of <em>Javanica</em> rice which are assumed feasible immediate progenitors of <em>Oryza sativa</em> [<a href=\"#r-9\">9</a>].Interestingly, the Assam center (nearby of Bangladesh) is also considered for the center of origin of such <em>Javanica </em>rice.<br />\r\nDay by day the number of rice genotypes become raise which ultimately harder the flexibility to differentiate of rice cultivars on the premise of morphological and biochemical attributes [<a href=\"#r-4\">4</a>]. Therefore several molecular markers such as SSR (Simple Sequence Repeat), RAPD (Random Amplified Polymorphic DNA), RFLP (Restriction Fragment Length Polymorphism), and AFLP (Amplified Fragment Length Polymorphism) etc. are widely utilized to be identified of particular cultivars [<a href=\"#r-10\">10</a>], or quality seed of hybrid varieties [<a href=\"#r-11\">11</a>] and for documentation of the released varieties in seed grain trade as well [<a href=\"#r-12\">12</a>]. Thus DNA fingerprinting data is additionally one of the example which is being globally practiced for the legal evidence of DUS (Distinctness, Uniformity, and Stability) [<a href=\"#r-13\">13</a>]–[<a href=\"#r-20\">20</a>].<br />\r\nPCR-based assays, co-dominant inheritance pattern, and elevated multi allelic variation/polymorphism are the major dynamic factors which steer the microsatellites/SSRs as the precious genetic markers of choice for the breeders [<a href=\"#r-21\">21</a>]–[<a href=\"#r-23\">23</a>]. The motif of the hyper variable microsatellites/SSRs are fairly located and abundantly well distributed all over the rice genome [<a href=\"#r-24\">24</a>]–[<a href=\"#r-28\">28</a>]. Hence, microsatellites/SSRs are also becoming efficient tools for the breeders as well as geneticists to incorporate genetic maps of rice having enormous wealth of diverge genetic variation [<a href=\"#r-29\">29</a>]–[<a href=\"#r-32\">32</a>]. Still, more than 50,000 microsatellites/SSRs have been designed in between <em>Indica </em>and <em>Japonica</em> rice accessions which are being contributed to construct genetic map for characterization and documentation of rice [<a href=\"#r-30\">30</a>],[<a href=\"#r-33\">33</a>]–[<a href=\"#r-36\">36</a>]. Based on the above scientific reports, such characterization and documentation process have been recently utilized in several varieties/landraces/cultivars/wild types of <em>Oryza sativa</em>, <em>Triticum aestivum</em>, <em>Zea mayes</em>, <em>Saccharum officinarum</em>, <em>Brassica napus</em>, <em>Glycine max</em>, <em>Solanum tuberosum</em>, <em>Corchorus capsularis</em>, and other crop species of Bangladesh [<a href=\"#r-13\">13</a>]–[<a href=\"#r-19\">19</a>],[<a href=\"#r-37\">37</a>]. However, in this study, the genetic fingerprinting techniques were utilized through five SSR markers for the protection of 110 local rice genotypes grown in Bangladesh. Moreover, the genetic fingerprinting techniques of this research can be further exploited for the protection and establishment of Intellectual Property Rights (IPR) of other crop species of Bangladesh.</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Raising of rice seedlings and extraction of DNA</strong><br />\r\nGenetic Resource and Seed (GSD) division of Bangladesh Rice Research Institute (BRRI) was provider of 108 cultivar’s seeds and the seeds of rest two cultivars named ACI-1 and Alok-932024 were supplied by ACI Pvt. Ltd of Bangladesh. Genetic fingerprinting lab of department of Genetics and Plant Breeding (GPB), Bangladesh Agricultural University (BAU), Bangladesh was being concerned to grow these seedlings as well as this experiment to be conducted. Genomic DNA of each cultivar was extracted from 2-5 inner succulent shoots of two-weeks-old fresh seedlings germinated in sterile petridishes. In that case, Rahman <em>et al</em>., [<a href=\"#r-13\">13</a>]–[<a href=\"#r-19\">19</a>],[<a href=\"#r-37\">37</a>],[<a href=\"#r-38\">38</a>] illustrated modified CTAB (cetyl trimethyl ammonium bromide) DNA extraction method supported by Aljanabi and Martinez [<a href=\"#r-39\">39</a>] was followed to isolate genomic DNA.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Quantification and optimization of DNA concentration</strong><br />\r\nUV-absorption spectrophotometer (Spectronic® Genesis™) was utilized at 260nm absorption to measure the purity and concentration of DNA which was subsequently estimated and converted into 25ng/µl with TE buffer, and finally stored at 4<sup>o</sup>C before to amplify with SSR primers.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Selection of polymorphic SSR markers for rice genotypes</strong><br />\r\nA total, 50 SSRs comprising on all the twelve chromosomes of rice were obtained. A set of seventeen SSR primers described previously [<a href=\"#r-38\">38</a>] were selected from them by surveying allelic polymorphism data from the available rice genome database (http://www.gramene.org) as illustrated by Rahman<em> et al.,</em> [<a href=\"#r-13\">13</a>]–[<a href=\"#r-19\">19</a>],[<a href=\"#r-37\">37</a>]. At first, three to five of those primers were tested through ten randomly selected genotypes setting with the recommended PCR thermal profile [<a href=\"#r-13\">13</a>]–[<a href=\"#r-19\">19</a>]. The expected ranges (base pairs length) PCR product was then validated based on the most excellent response to amplify the target genomic region of the template DNA. In such a way five SSR primers <em>viz</em>., RM153, RM251, RM333, RM335 and RM475 which depict 5, 3, 10, 4, and 2 of rice chromosome [<a href=\"#r-36\">36</a>] were preferred by visualizing clear and predictable amplified alleles, and finally employed for SSR analysis in this study (Figure 1). The selected primers were subsequently run with all 110 cultivars at once which displayed clear and repeatable polymorphic bands.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Polymerase chain reaction (PCR) amplification profile for SSRs</strong><br />\r\nEppendorf<sup>™</sup> oil-free thermal cycler gradient was utilized in this PCR reaction. Approximate 15 μl reaction volume consists of 50ng sample DNA (2.0 µl), 10x PCR Buffer (3.0 µl), 1 µM of each forward and reverse SSR primer (1.0 µl), 0.25 mM dNTPs (1.5 µl), 1 unit ampli Taq DNA polymerase (0.5 µl), and nuclease free double distilled deionized water (6.0 µl) were utilized to perform PCR reaction. PCR settings were carried out by the conditions described by Panaud <em>et al.,</em> [<a href=\"#r-34\">34</a>],[<a href=\"#r-36\">36</a>] with minor modifications suggested by previous research [<a href=\"#r-13\">13</a>]–[<a href=\"#r-19\">19</a>],[<a href=\"#r-37\">37</a>] as follows: 95°C for 5 min (an initial denaturation) followed by entire 35 cycles, 95°C for 40 sec (denaturation), 55°C for 30 sec (annealing) and 72°C for 1 min (elongation/extension), then a final elongation/extension cycle at 72°C for 7 min. Amplified PCR reactions were then stored at –20<sup>o</sup>C for further utilization.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Electrophoresis and inspection of banding patterns</strong><br />\r\nTop vision submarine horizontal electrophoresis system (BIORAD<sup>™</sup> Sequencing Cell) was utilized to electrophoresis the PCR reactions (Figure 1). Prior to electrophoresis, each 07 µL amplified PCR aliquot and 3 µL of loading buffer [38] were mixed gently. This mixture was then loaded on 2% agarose gel, and placed into the submarine horizontal gel chamber with 1x TBE running buffer (Trizma base, boric acid and EDTA; pH 8.0). A five microlitre (5 µL) 100 bp standard DNA (Gene ruler, Fermentas<sup>®</sup>) ladder was added in both left and right side of the gel to compare the molecular weight of the amplified PCR products of each cultivar. PCR mixtures (10 <em>μ</em>l) were subjected to electrophoresis at 100V and 50W for 2 hrs 40 mins. The electrophoresis sample was then kept as photographic image by the camera polaroid gel documentation system (UVP, BioDoc-It<sup>™ </sup>System).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Genotyping of alleles and data analysis</strong><br />\r\nDNA FRAG v3.03 computer software [<a href=\"#r-39\">39</a>] was utilized to measure the most profoundly amplified alleles per loci (Figure 1) by using 100bp recognizing size standard DNA ladder [<a href=\"#r-40\">40</a>]. The individual unambiguous DNA fragments were referred as alleles of the respective SSR markers. The allele frequency data (DNA fragment) was exported as diploid datasheet arrangement (AA, AB, CC and so on) on POPGENE v1.31 computer program [<a href=\"#r-42\">42</a>], and therefore, utilized for the several statistical analysis including “observed number of alleles-N<em>a</em> [<a href=\"#r-42\">42</a>]”, “effective number of alleles-N<em>e</em>” [<a href=\"#r-43\">43</a>], “allelic diversity index (PIC=1-∑X<sub>i</sub><sup>2</sup>, where X<sub>i</sub> indicates the frequency of the i<sup>th</sup> allele), Shannon’s Information index-<em>I</em> [<a href=\"#r-44\">44</a>]”, “Hardy-Weinberg equilibrium (H<em>e</em> and H<em>o</em> of Levene [<a href=\"#r-45\">45</a>] and Gene flow-N<sub>m</sub>)”, “Nei’s gene diversity index (<em>h</em>) [<a href=\"#r-46\">46</a>]”, “chi-square & probability index”, and “Wright’s fixation index-F<sub>is </sub>[<a href=\"#r-47\">47</a>]”. POPGENE v1.31 software was also applied to estimate genetic distance and similarity among the genotypes. Finally, an UPGMA (Unweighted Pair Group Method of Arithmetic Means) phylogenetic tree (Figure 2) was assembled by means of Nei’s [<a href=\"#r-48\">48</a>] genetic distance (<em>D</em>) which was visualized via Treeview computer software [<a href=\"#r-49\">49</a>]. The generated cluster on UPGMA diagram (Figure 2) was then used to explain the relationships among the cultivars in this study.</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><strong>Allele frequency and allelic diversity index (PIC)</strong><br />\r\nUsing 5 polymorphic loci in 110 cultivars of rice, a complete of 99 alleles were found in this quantitative investigation where RM335 revealed the foremost observed alleles (25) followed by RM333 (21), RM251 (20), RM475 (19) and RM153 (14) as shown in <a href=\"#Table-1\">Table 1</a>. The Highest effective number (N<em>e</em>) of alleles (19.852) was also found in RM335 (<a href=\"#Table-2\">Table 2</a>). The diversity index of alleles or Polymorphism Information Content (PIC=1-∑Xi<sup>2</sup>) values usually reflect particular allele diversity into a species [<a href=\"#r-50\">50</a>]. The average PIC value was 0.864 with the number ranging from 0.879 (RM153) to 0.949 (RM335) (<a href=\"#Table-1\">Table 1</a>).</p>\r\n\r\n<div id=\"Table-1\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1588348425-table1/\">Table-1</a><strong>Table 1.</strong> Fingerprinting Alleles and polymorphism information content (PIC) of five SSR loci across 110 rice cultivars.</p>\r\n</div>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Genetic variation statistics</strong><br />\r\nAspect of allele frequency of each cultivars and total cultivars were taken into consideration to calculate the comprehensive Shannon’s Information Index (<em>I</em>) which is fluctuated from 2.284 (RM153) to 3.080 (RM335). Fixation index (F<sub>is</sub>) (a measure of genetic differentiation) was recorded from 0.687 to 0.958 having 0.838 average values (<a href=\"#Table-2\">Table 2</a>). Highest gene flow (N<sub>m</sub>) was estimated through RM251 microsatellite loci (0.046) while RM153 showed the lowest gene flow (0.005) in this study (<a href=\"#Table-2\">Table 2</a>).</p>\r\n\r\n<p>Across 110 rice varieties, RM251 (0.290) yielded the very best average heterozygosity (H<em><sub>O</sub></em>) in current study followed by RM475 (0.172), RM335 (0.154), RM333 (0.090) and RM153 (0.036) (<a href=\"#Table-2\">Table 2</a>). Highest heterozygosity can be explained as a result of length and distance of RM251 marker on the genetic map relative to centromere [<a href=\"#r-14\">14</a>].</p>\r\n\r\n<div id=\"Table-2\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1588348425-table2/\">Table-2</a><strong>Table 2.</strong> Summary statistics of the utilized genetic diversity parameters in the study.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Varietal identifications</strong><br />\r\nComparative SSR profiles and DNA molecular weight (band position) against five SSR primers (<a href=\"#figure1\">Figure 1</a>), all the varieties were distinguished from each other with a minimum of single and/or arrangement of five primers (<a href=\"#Table-3\">Table 3</a>).</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"542\" src=\"/media/article_images/2024/38/14/178-1588348425-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1</strong>. A) Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 1 to 28 represents rice cultivars (1=Lal Amon, 2=Lau Jan, 3=Nara Aswina, 4=Buna Dhan, 5=Begun Bechi, 6=Bhasha Manik, 7=Kochu Dhola, 8=Sunga Wala, 9=Konek Chul, 10=Horinkhur Panati, 11=Ganjia, 12=Dudhsar, 13=Mathia, 14=Khirma Pat, 15=Mukut Sail, 16=Ban Kolom, 17=Safa Har (3), 18=Kal Nania, 19=Shil Pan, 20=Jabar Sail, 21=Moisha Mida, 22=Paglakushyari, 23=Pan Kaich, 24=Lal Patjat, 25=Chandda Gotok, 26=Moisha Mira, 27=Choia Mora, 28=Modhu Maloti) [38]. B) B. Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 29 to 56 represents rice cultivars (29=Kali Gochya, 30=Ludi Gochya, 31=Kala Gura, 32=Monura, 33=Moisa Mira, 34=Chand Moni (3), 35=Kamoni Sail, 36=Juna, 37=Kurki, 38=Arai Raj, 39=Kala Gora, 40=Mulai, 41=Dharga Sail, 42=Bondyl, 43=Chakkol, 44=Rajamun, 45=Kanchon Mogi, 46=Hiruyal, 47=Chapa Mali, 48=Deppol, 49=Pushon, 50=Kali Cochr, 51=Ghori Amon, 52=Muirol, 53=Muar Sail, 54=Jhaw Lota, 55=Bhoban, 56=Ful Badam), C) Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 57 to 84 represents rice cultivars (57=Galong, 58=Hasna Chikon, 59=Thakor, 60=Tembur, 61=Tulsi Mala, 62=Giring, 63=Nag Pechi, 64=Bhua Dhan, 65=Sandik Sail, 66=Jhoria Sail, 67=Halde Medi, 68=Maitya Cheng, 69=Kala Bail, 70=Murki Balam, 71=Gulchamlaish, 72=Raj Kumari, 73=Hati Banda, 74=Kui Sail (2), 75=Chakkol (Muta) Raozan, 76=Muijuri, 77=Munsi Sail, 78=Thakur Dhan, 79=Moina Sail, 80=Butu Balam(2), 81=Kali Jira (2), 82=Jhual Kata, 83=Raj Bhog, 84=Lal Modonga), and D) Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 85 to 110 represents rice cultivars (85=Surma Sail, 86=Madhu Mala, 87=Dumai Sail, 88=Chadlash, 89=Jola Bhangh, 90=Mutonga (2), 91=Jula Gudi, 92=Gutok, 93=Pani Torong, 94=Randar, 95=Bhor Gelam, 96=Neel Huri, 97=Jol Kumari, 98=Ful Kari, 99=Biropa, 100=Asham Baba, 101=Dhul Abiz, 102=Porangi, 103=Tulo Sail, 104=Khoni Dhan, 105=Choro, 106=Sundar Sail, 107=Bokra, 108=Nagra, 109=ACI 1, 110=Aalok 932024).</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"Table-3\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1588348425-table3/\">Table-3</a><strong>Table 3. </strong>Distinction of 110 rice cultivars all the way through SSR band positions.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Analysis of genetic distance and phylogenetic tree</strong><br />\r\nThe summary of Nei’s genetic distance (<em>D</em>) from 5995 varietal pairs among 110 rice cultivars varied from zero to 2.832. Out of these varietal pairs, 58.87% (3529) showed no genetic distance [<a href=\"#r-11\">11</a>]. Such genetic distance and similarity in this study separated all the 110 cultivars into several clusters (“a” to “x”) at once on the UPGMA diagram (<a href=\"#figure2\">Figure 2</a>).</p>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"630\" src=\"/media/article_images/2024/38/14/178-1588348425-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2.</strong> UPGMA phylogenetic tree based on Nei’s [63] genetic distance showing the genetic relationship among 110 rice cultivars (Group, A= Transplant Aman, B= Broadcast Aman, C= Boro and Jhum)</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>All the utilized polymorphic SSR markers recorded a complete of 99 unique alleles (Table 1) which was significantly higher than the total number of alleles reported by the several previous researches [<a href=\"#r-13\">13</a>]–[<a href=\"#r-19\">19</a>],[<a href=\"#r-33\">33</a>]. On an average it yielded 19.80 alleles per primer with an effect of 38.81%. In agreement with earlier works [<a href=\"#r-13\">13</a>]–[<a href=\"#r-19\">19</a>],[<a href=\"#r-33\">33</a>] reported a total of 18 alleles [<a href=\"#r-16\">16</a>],[<a href=\"#r-17\">17</a>] through analyzing with three primers (RM11; RM151 and RM153) and 78 [<a href=\"#r-15\">15</a>] alleles with five primers (RM1; RM151; RM153; RM334 and RM335), respectively, while running on diverse ecotypes of rice genotypes of Bangladesh from the protected rice materials of the BRRI. In those investigations, PIC values were recorded 0.670; 0.707; 0.698 [<a href=\"#r-16\">16</a>],[<a href=\"#r-17\">17</a>] & 0.862; 0.923; 0.831; 0.865 and 0.910, respectively [<a href=\"#r-15\">15</a>]. In another study, a total of 238 rice accessions (<em>Indica </em>and <em>Japonica)</em> by using entire ten microsatellite markers were investigated by Yang and his associates [<a href=\"#r-51\">51</a>] where they observed maximum 25 exclusive alleles. Genotypes under the low PIC value study represent closely related variants, while superior PIC indicates considerable enormous diversity, which is ideal for the development of new variants as well [<a href=\"#r-52\">52</a>]. The frequency of short tandem repeats of microsatellites as well as their repeat sequences have a command on the quantity of amplified alleles and their resultant PIC values of the experimental genotypes [<a href=\"#r-35\">35</a>],[<a href=\"#r-36\">36</a>],[<a href=\"#r-53\">53</a>],[<a href=\"#r-54\">54</a>]. In addition, Ni <em>et al</em>., revealed from his investigation that more extensive repeats including GA- sequenced repeats acquiesce more quantity of distinctive alleles with superior PIC standards [<a href=\"#r-54\">54</a>]. In contrast, it has been suggested that the motif of (CTT)<em><sub>n</sub> </em>and amplified AT-affluent trinucleotide repeats also exhibits adequate and greater polymorphism of alleles [<a href=\"#r-35\">35</a>]. RM333 primer containing (CTT)<em><sub>n</sub> </em>motif was one of the most instructional SSR marker because it gave 6-7 distinctive alleles and standard PIC range in Temnykh’s experiment [<a href=\"#r-36\">36</a>]. In support of RM335 [(CTT)<sub>20</sub>] SSR primers, 25 unique alleles with 0.910 PIC numeral were recorded which were the foremost alleles and the maximum PIC numeral in this investigation. The PIC valuation is considered as the discriminating strength of a promising marker to the genetic diversity study of the breeding materials selection program for the breeders because it regulates the frequency of observed and effective alleles of a particular DNA marker [<a href=\"#r-55\">55</a>]–[<a href=\"#r-57\">57</a>]. However, remarkably elevated PIC (0.879 to 0.949) values in this investigation indicated that the chosen markers have the required properties to be used in this DNA fingerprinting research among the 110 rice cultivars grown in Bangladesh [<a href=\"#r-58\">58</a>]. However, the observation of this study was partially supported by the points of accuracy and usefulness from the above discussions.<br />\r\nLocation of particular DNA genetic markers on the precise chromosomes, frequencies and size of alleles through their PIC numeral are given in <a href=\"#Table-1\">Table 1</a>. Mutation and chromosomal crossover are the two general events of heredity which are usually occupied at distal proximity from the centromere of the chromosome. They usually effort the formation of abundant alleles and eventually diversity of a specific locus [<a href=\"#r-13\">13</a>]–[<a href=\"#r-16\">16</a>],[<a href=\"#r-18\">18</a>],[<a href=\"#r-19\">19</a>],[<a href=\"#r-37\">37</a>]. 24.7cM (chromosome 5); 79.1 cM (chromosome 3); 110.4cM (chromosome 10); 21.5cM (chromosome 4) and 92.5cM (chromosome 2) are the located positions of RM153; RM251; RM333; RM335 and RM475 primers on rice chromosome [<a href=\"#r-36\">36</a>]. These primers were applied in the current observation of 110 rice cultivars including 2 hybrids and one variety of Jhum cultivation system. Overall gene flow values and genetic diversity were observed in several populations of <em>Oryza officinalis</em> and computed 0.316 and 0.442, respectively, by examining of entire 14 microsatellite markers [<a href=\"#r-59\">59</a>]. A wide range of allele frequency along with PIC value, major genetic variation in sense of observed and expected heterozygosity (H<em>o</em> and H<em>e</em>) were detected in this investigation. The observed and expected heterozygosity (H<em>o</em> and H<em>e</em>) values were estimated from 0.036 (RM155) to 0.290 (RM 251), and from 0.883 (RM155) to 0.954 (RM335), respectively. Superior expected heterozygosity (H<em>e</em>) content than the observed heterozygosity in this study indicated that the selected SSR markers were remarkably abundant informative for the DNA fingerprinting among the rice cultivars [<a href=\"#r-60\">60</a>],[<a href=\"#r-61\">61</a>]. Partial consistent in theses’ observations were computed in several Bangladeshi local rice cultivars by a number of previous research groups [<a href=\"#r-4\">4</a>],[<a href=\"#r-17\">17</a>],[<a href=\"#r-38\">38</a>]. Greater genetic variation and a lower gene flow value in 110 rice varieties argued that the most studied varieties in this experiment were landraces [<a href=\"#r-38\">38</a>].<br />\r\nIn these research materials, a total of 92, 15, 02 and 01 cultivars are recommended for transplant aman (T. Aman), broadcast aman (B. Aman), boro and jhum ecotype by BRRI [<a href=\"#r-14\">14</a>]. In diallel fashion, a total of 5995 varietal pairs were possible among 110 cultivars where 2466 (41.13%) varietal pairs were computed as to be prominent genetic distance (Nei’s genetic distance-<em>D</em>) with each other [<a href=\"#r-14\">14</a>]. In a previous study, while analyzing 94 varieties of six different ecotypes of rice grown in Bangladesh, a total of 4371 varietal pairs were computed of which 37% appeared nil genetic distance, and merely 1% showed highest genetic distance (2.583) [<a href=\"#r-15\">15</a>],[<a href=\"#r-62\">62</a>]. This closeness may be possible due to the genetic make-up of the locus for which the primers were responsible to distinguish along with low ecotype variation. Among the 5995 varietal pairs, only 0.917% appeared highest (2.832) genetic distance in this research. However, the superior genetic distance (<em>D</em>) is often observed while the cultivars or any genotypes were occupied from the landraces or wild relatives, in one side, and the high-yielding varieties (HYVs) on the opposite side in their crossing events [<a href=\"#r-38\">38</a>]. Thus, the variation between highest and lowest genetic distance among the 110 cultivars proved their existence of variability. The resulting such genetic variability of the cultivars can be applied as a parent material in the future variety improvement programs to seek out the most efficient cultivars for further crossing or breeding.<br />\r\nHowever, Nei’s genetic distance (<em>D</em>) while analyzed on the UPGMA dendrogram considering 110 cultivars at a time, the dendogram separated the varieties, Dhul Abiz, Biropa and Bhor Gelam (Cluster “a”) from other 107 cultivars (Cluster “b”). Cluster “b” subsequently separated into sub-cluster “c” (Dumai Sail and Jol kumari) and sub-cluster “d” containing other 105 rice varieties. Sub-cluster subsequently formed other sub-clusters namely, “e”, “f”, “g”, “h” and so on (<a href=\"#figure2\">Figure 2</a>). The varieties, as for example, Jol Kumari, Sandik sail, Jhul Kata, Thakor, Aalok 932024 and Tulu Sail were found in different sub-clusters “c”, “g”, “i”, “o”, “w” and “x”, respectively, due to their genetic distance. The major sub-clusters (“u”-“x”) were found to cover 39 of the 110 cultivars starting from ACI 1 to Lal Amon, all of which are traditional rice varieties of Bangladesh except ACI 1 and Aalok 932024. UPGMA dendrogram within the groups; Transplant Aman (A), Broadcast Aman (B), Boro and Jhum (C) are given in <a href=\"#figure2\">Figure 2</a>. As two Boro varieties (ACI 1 and Aalok 932024) and one Jhum variety (Ful Badam) were used in this experiment it was not possible to analyze them individually: they were therefore combined for a single analysis. An attempt was made to distinguish the varieties as their ecotype situations. The UPGMA dendrogram was constructed for this purpose (<a href=\"#figure2\">Figure 2</a>). The results showed that the ecotypes have distinct clusters to represent Jhum and BRRI Accessions have formed the unique cluster different from all others as expected. The groups Transplant Aman (T. Aman) and Broadcast Aman (B. Aman) formed two closely linked sub-sub-clusters under one sub-cluster, while showing distinct difference from these sub-groups. Jhum formed a unique cluster and Boro a sub-cluster.<br />\r\nIn these 110 cultivars, all the cultivars were distinguished from one another with either through 1<sup>st</sup>, 2<sup>nd</sup>, 3<sup>rd</sup>, 4<sup>th</sup>, and 5<sup>th</sup> SSR primers (<a href=\"#Table-3\">Table 3</a>) and also through qualitative and quantitative traits of Breeders [<a href=\"#r-18\">18</a>]. Many varieties had similar names, which had created a number of problems related to final characterization. The varieties Moisha Mida (T. Aman), Moisha Mira (T. Aman), Moisa Mira (B. Aman); Kala Gura (B. Aman), Kala Gora (T. Aman) and Thakor (T. Aman), Thakur Dhan (T. Aman) have similar names but when studied by both qualitative and genetic fingerprinting those showed distinct differences [<a href=\"#r-14\">14</a>].</p>"
},
{
"section_number": 5,
"section_title": "CONCLUSIONS",
"body": "<p>In this study, the registered local rice cultivars grown in Bangladesh were exploited to distinct each and every rice cultivar based on identification of the DNA band patterns by means of specific primers, generally termed as genetic fingerprinting. All the utilized cultivars in this research were distinguished from one another with either 1<sup>st</sup>, 2<sup>nd</sup>, 3<sup>rd</sup>, 4<sup>th</sup>, or 5<sup>th</sup> SSRs. It is important to note that some of the varieties of traditional types had very similar names, which usually gave an understanding of repeats, but it was interestingly found to be distinctly different from one another due to molecular data. An example is the varieties named Moisa Mira, Moisha Mira, Moisha Mida, Thakor, Thakur Dhan, both pairs of which were distinct from one another on molecular traits. Therefore, these were not repeated but individually distinct cultivars or land races of rice collected from different source areas of Bangladesh by BRRI scientists at different times. However, Intellectual Property Rights (IPR) and Plant Variety Protection (PVP) of wild relatives, landraces, cultivars and commercial varieties of Bangladeshi rice will be guided from the outcomes of this research. Such series of works on more rice genetic materials as well as other crop species should be done as a regular study by the genetic resource centers of different institutes in collaboration with universities, where government should give adequate financial and special manpower support with appropriate incentives for those who will lead the program.</p>"
},
{
"section_number": 6,
"section_title": "ACKNOWLEDGEMENTS",
"body": "<p>Authors express thanks to the DANIDA for supporting this research work through the Agriculture Extension Component/Seed Wing of the Ministry of Agriculture, Government of the People’s Republic of Bangladesh. In the same line, they extend thanks to Mr. Anwar Faruqe, Former Joint Secretary MoA and Former Director General of the Seed Wing, for his support and keen interest in the project activities.</p>"
},
{
"section_number": 7,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>LR, MSEA and MKB were involved in conception and design of the experiments. MSR and MKHS contributed to perform the experiments. LR, UKN, MSR and MKHS contributed to drafting the article. LR and MSR contributed to revising it critically for important intellectual content. MSR made the final approval of the version to be published.</p>"
},
{
"section_number": 8,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/38/14/178-1588348425-Figure1.jpg",
"caption": "Figure 1. A) Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 1 to 28 represents rice cultivars (1=Lal Amon, 2=Lau Jan, 3=Nara Aswina, 4=Buna Dhan, 5=Begun Bechi, 6=Bhasha Manik, 7=Kochu Dhola, 8=Sunga Wala, 9=Konek Chul, 10=Horinkhur Panati, 11=Ganjia, 12=Dudhsar, 13=Mathia, 14=Khirma Pat, 15=Mukut Sail, 16=Ban Kolom, 17=Safa Har (3), 18=Kal Nania, 19=Shil Pan, 20=Jabar Sail, 21=Moisha Mida, 22=Paglakushyari, 23=Pan Kaich, 24=Lal Patjat, 25=Chandda Gotok, 26=Moisha Mira, 27=Choia Mora, 28=Modhu Maloti) [38]. B) B. Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 29 to 56 represents rice cultivars (29=Kali Gochya, 30=Ludi Gochya, 31=Kala Gura, 32=Monura, 33=Moisa Mira, 34=Chand Moni (3), 35=Kamoni Sail, 36=Juna, 37=Kurki, 38=Arai Raj, 39=Kala Gora, 40=Mulai, 41=Dharga Sail, 42=Bondyl, 43=Chakkol, 44=Rajamun, 45=Kanchon Mogi, 46=Hiruyal, 47=Chapa Mali, 48=Deppol, 49=Pushon, 50=Kali Cochr, 51=Ghori Amon, 52=Muirol, 53=Muar Sail, 54=Jhaw Lota, 55=Bhoban, 56=Ful Badam), C) Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 57 to 84 represents rice cultivars (57=Galong, 58=Hasna Chikon, 59=Thakor, 60=Tembur, 61=Tulsi Mala, 62=Giring, 63=Nag Pechi, 64=Bhua Dhan, 65=Sandik Sail, 66=Jhoria Sail, 67=Halde Medi, 68=Maitya Cheng, 69=Kala Bail, 70=Murki Balam, 71=Gulchamlaish, 72=Raj Kumari, 73=Hati Banda, 74=Kui Sail (2), 75=Chakkol (Muta) Raozan, 76=Muijuri, 77=Munsi Sail, 78=Thakur Dhan, 79=Moina Sail, 80=Butu Balam(2), 81=Kali Jira (2), 82=Jhual Kata, 83=Raj Bhog, 84=Lal Modonga), and D) Representative gel pictures of amplified alleles from five SSR markers RM153, RM251, RM333, RM335 and RM475 by using several rice cultivars of Bangladesh (Lane M=100bp Ladder; Lanes 85 to 110 represents rice cultivars (85=Surma Sail, 86=Madhu Mala, 87=Dumai Sail, 88=Chadlash, 89=Jola Bhangh, 90=Mutonga (2), 91=Jula Gudi, 92=Gutok, 93=Pani Torong, 94=Randar, 95=Bhor Gelam, 96=Neel Huri, 97=Jol Kumari, 98=Ful Kari, 99=Biropa, 100=Asham Baba, 101=Dhul Abiz, 102=Porangi, 103=Tulo Sail, 104=Khoni Dhan, 105=Choro, 106=Sundar Sail, 107=Bokra, 108=Nagra, 109=ACI 1, 110=Aalok 932024).",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/38/14/178-1588348425-Figure2.jpg",
"caption": "Figure 2. UPGMA phylogenetic tree based on Nei’s [63] genetic distance showing the genetic relationship among 110 rice cultivars (Group, A= Transplant Aman, B= Broadcast Aman, C= Boro and Jhum)",
"featured": false
}
],
"authors": [
{
"id": 850,
"affiliation": [
{
"affiliation": "Department of Biotechnology, Faculty of Agriculture Patuakhali Science and Technology University, Dumki, Patuakhali-8602, Bangladesh."
}
],
"first_name": "Md Shafikur",
"family_name": "Rahman",
"email": "shafikjss@gmail.com",
"author_order": 1,
"ORCID": null,
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Md Shafikur Rahman, E-mail: shafikjss@gmail.com, Phone: +8801717469112",
"article": 193
},
{
"id": 851,
"affiliation": [
{
"affiliation": "Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh."
}
],
"first_name": "Md Kamrul Hasan",
"family_name": "Sohag",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 193
},
{
"id": 852,
"affiliation": [
{
"affiliation": "Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh."
}
],
"first_name": "Lutfur",
"family_name": "Rahman",
"email": null,
"author_order": 3,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 193
},
{
"id": 853,
"affiliation": [
{
"affiliation": "Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh."
}
],
"first_name": "Md Shah-E-",
"family_name": "Alam",
"email": null,
"author_order": 4,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 193
},
{
"id": 854,
"affiliation": [
{
"affiliation": "Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh."
}
],
"first_name": "Ujjal Kumar",
"family_name": "Nath",
"email": null,
"author_order": 5,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 193
},
{
"id": 855,
"affiliation": [
{
"affiliation": "Genetic Resource & Seed Division, Bangladesh Rice Research Institute, Joydebpur, Gazipur, Bangladesh"
}
],
"first_name": "Md. Khairul",
"family_name": "Bashar",
"email": null,
"author_order": 6,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 193
}
],
"views": 420,
"downloads": 81,
"references": [
{
"id": 6492,
"serial_number": 1,
"pmc": null,
"reference": "Wennberg A. Food and Agriculture Organization of the United Nations. In: Encyclopedia of Toxicology: Third Edition. 2014. DOI: 10.1016/B978-0-12-386454-3.00988-X",
"DOI": null,
"article": 193
},
{
"id": 6493,
"serial_number": 2,
"pmc": null,
"reference": "Anonymous. National Workshop on Rice Research and Extension-2002. Feeding the extra millions by 2025. Bangladesh Rice Research Institute Gazipur. 2002:1.",
"DOI": null,
"article": 193
},
{
"id": 6494,
"serial_number": 3,
"pmc": null,
"reference": "Jackson MT. Protecting the heritage of rice biodiversity. Geo Journal. 1995; 35(3): 267–274. DOI: 10.1007/BF00989134",
"DOI": null,
"article": 193
},
{
"id": 6495,
"serial_number": 4,
"pmc": null,
"reference": "Siddique MA, Khalequzzaman M, Islam MM, Fatema K, Latif MA. Molecular characterization and genetic diversity in geographical indication (GI) rice (Oryza sativa L.) cultivars of Bangladesh. Revista Brasileira de Botanica. 2016; 39(2): 631–640. DOI: 10.1007/s40415-016-0271-1",
"DOI": null,
"article": 193
},
{
"id": 6496,
"serial_number": 5,
"pmc": null,
"reference": "Nadir S, Xiong HB, Zhu Q, Zhang XL, Xu HY, Li J, et al. Weedy rice in sustainable rice production. A review. Agron Sustain Dev. Agronomy for Sustainable Development. 2017; Springer-Verlag France. DOI: 10.1007/s13593-017-0456-4",
"DOI": null,
"article": 193
},
{
"id": 6497,
"serial_number": 6,
"pmc": null,
"reference": "Qiu J, Zhou Y, Wang Y, Mao L, Ye C, Wang W, et al. Genomic variation associated with local adaptation of weedy rice during de-domestication. Nature Communication. 2017; 8: 15323. DOI: 10.1038/ncomms15323",
"DOI": null,
"article": 193
},
{
"id": 6498,
"serial_number": 7,
"pmc": null,
"reference": "Zhao Z. New data and new issues for the study of origin of rice agriculture in China. Archaeological and Anthropological Sciences. 2010; 2(2): 99-105. DOI: 10.1007/s12520-010-0028-x",
"DOI": null,
"article": 193
},
{
"id": 6499,
"serial_number": 8,
"pmc": null,
"reference": "Choi JY, Platts AE, Fuller DQ. The rice paradox: Multiple origins but single domestication in Asian Rice. Molecular Biology and Evolution. 2017; 34(4): 969-979. DOI: 10.1093/molbev/msx049",
"DOI": null,
"article": 193
},
{
"id": 6500,
"serial_number": 9,
"pmc": null,
"reference": "Chang TT. The origin, evolution, cultivation, dissemination, and diversification of Asian and African rices. Euphytica. 1976; 25(1): 425-441. DOI: 10.1007/BF00041576",
"DOI": null,
"article": 193
},
{
"id": 6501,
"serial_number": 10,
"pmc": null,
"reference": "Mailer RJ, Scarth R, Fristensky B. Discrimination among cultivars of rapeseed (Brassica napus L.) using DNA polymorphisms amplified from arbitrary primers. Theoretical and Applied Genetics: International Journal of Plant Breeding Research. 1994; 87(6): 697-704. DOI: 10.1007/BF00222895",
"DOI": null,
"article": 193
},
{
"id": 6502,
"serial_number": 11,
"pmc": null,
"reference": "Marshall P, Marchand MC, Lisieczko Z, Landry BS. A simple method to estimate the percentage of hybridity in canola (Brassica napus) F1 hybrids. Theoretical and Applied Genetics. 1994; 89(7-8): 853-858. DOI: 10.1007/BF00224508",
"DOI": null,
"article": 193
},
{
"id": 6503,
"serial_number": 12,
"pmc": null,
"reference": "Bligh HFJ, Blackhall NW, Edwards KJ, McClung AM. Using amplified fragment length polymorphisms and simple sequence length polymorphisms to identify cultivars of brown and white milled rice. In: Crop Science; 1999: 39(6); 1715-1721. DOI: 10.2135/cropsci1999.3961715xer",
"DOI": null,
"article": 193
},
{
"id": 6504,
"serial_number": 13,
"pmc": null,
"reference": "Rahman L, Nath UK, Molla MR, Islam MN, Siddique MA, Rahman MS, et al. Plant Varieties of Bangladesh: Morphological and Molecular Characterization. Vol 4. Seed Wing, Ministry of Agriculture, Government of the People’s Republic of Bangladesh; 2010; 4: 550.",
"DOI": null,
"article": 193
},
{
"id": 6505,
"serial_number": 14,
"pmc": null,
"reference": "Rahman L, Rahman MS, Nath UK, Bashar MK, Sohag MKH. Plant Varieties of Bangladesh: Morphological and Molecular Characterization Vol 3. Published by Seed Wing, Ministry of Agriculture, Government of the People’s Republic of Bangladesh. 2009; 3: 394. DOI: 10.13140/RG.2.1.3062.6640",
"DOI": null,
"article": 193
},
{
"id": 6506,
"serial_number": 15,
"pmc": null,
"reference": "Rahman L, Rahman MS, Islam MN, Islam MS. Rahman L, Islam MN, Rahman MS, Islam MS. Plant Varieties of Bangladesh: Morphological and Molecular Characterization. Vol 2. Published by Seed Wing, Ministry of Agriculture, Govt of the People’s Republic of Bangladesh; 2008; 2: 300. https://www.researchgate.net/publication/286146036_PLANT_VARIETIES_OF_BANGLADESH_Morphological_and_Molecular_Characterization_Volume-2 .",
"DOI": null,
"article": 193
},
{
"id": 6507,
"serial_number": 16,
"pmc": null,
"reference": "Rahman L, Molla MR, Sultana S, Islam MN, Ahmed NU, Rahman MS, et al. PLANT VARIETIES OF BANGLADESH: Morphological and Molecular Characterization. Vol 1. Published by Seed Wing, Ministry of Agriculture, Government of the Peoples’ Republic of Bangladesh; 2007; 1: 486. DOI: 10.13140/RG.2.1.3849.0968",
"DOI": null,
"article": 193
},
{
"id": 6508,
"serial_number": 17,
"pmc": null,
"reference": "Rahman MS, Molla MR, Alam MS, Rahman L. DNA fingerprinting of rice (Oryza sativa L.) cultivars using microsatellite markers. Australian Journal of Crop Science. 2009; 3(3): 122-128. ISSN: 18352693",
"DOI": null,
"article": 193
},
{
"id": 6509,
"serial_number": 18,
"pmc": null,
"reference": "Rahman M, Sohag M, Rahman L. Distinctness of 110 rice (Oryza sativa L.) varieties of Bangladesh through morphological traits. Journal of the Bangladesh Agricultural University. 2014; 12(1): 29-36. DOI: 10.3329/jbau.v12i1.21236",
"DOI": null,
"article": 193
},
{
"id": 6510,
"serial_number": 19,
"pmc": null,
"reference": "Alam MK, Rahman MS, Mustakima QKJ, Sohag MKH, Ahmed NU, Nath UK, et al. SSR based molecular diversity study in local rice (Oryza sativa L.) genotypes of Bangladesh. International Journal of Innovative Research. 2016; 1(2): 17–24. http://www.irsbd.org/welcome/full_article/29",
"DOI": null,
"article": 193
},
{
"id": 6511,
"serial_number": 20,
"pmc": null,
"reference": "Ahmed MSU, Khalequzzaman M, Bashar MK, Shamsuddin AKM. Agro-Morphological, Physico-Chemical and Molecular Characterization of Rice Germplasm with Similar Names of Bangladesh. Rice Science. 2016. DOI: 10.1016/j.rsci.2016.06.004",
"DOI": null,
"article": 193
},
{
"id": 6512,
"serial_number": 21,
"pmc": null,
"reference": "Thomson MJ, Septiningsih EM, Suwardjo F, Santoso TJ, Silitonga TS, McCouch SR. Genetic diversity analysis of traditional and improved Indonesian rice (Oryza sativa L.) germplasm using microsatellite markers. Theoretical and Applied Genetics. 2007; 114(3): 559–568. DOI: 10.1007/s00122-006-0457-1",
"DOI": null,
"article": 193
},
{
"id": 6513,
"serial_number": 22,
"pmc": null,
"reference": "Kalia RK, Rai MK, Kalia S, Singh R, Dhawan AK. Microsatellite markers: An overview of the recent progress in plants. Euphytica. 2011. DOI: 10.1007/s10681-010-0286-9",
"DOI": null,
"article": 193
},
{
"id": 6514,
"serial_number": 23,
"pmc": null,
"reference": "Nadeem MA, Nawaz MA, Shahid MQ, Doğan Y, Comertpay G, Yıldız M, et al. DNA molecular markers in plant breeding: current status and recent advancements in genomic selection and genome editing. Biotechnology and Biotechnological Equipment. Taylor and Francis Ltd.. 2018. DOI: 10.1080/13102818.2017.1400401",
"DOI": null,
"article": 193
},
{
"id": 6515,
"serial_number": 24,
"pmc": null,
"reference": "Akagi H, Yokozeki Y, Inagaki A, Fujimura T. Microsatellite DNA markers for rice chromosomes. Theoretical and Applied Genetics. 1996; 93(7): 1071-1077. DOI: 10.1007/BF00230127",
"DOI": null,
"article": 193
},
{
"id": 6516,
"serial_number": 25,
"pmc": null,
"reference": "McCouch SR, Chen X, Panaud O, Temnykh S, Xu Y, Cho Y, et al. Microsatellite marker development, mapping and applications in rice genetics and breeding. Plant Molecular Biology. 1997; 35(1-2): 89-99. DOI: 10.1007/978-94-011-5794-0_9",
"DOI": null,
"article": 193
},
{
"id": 6517,
"serial_number": 26,
"pmc": null,
"reference": "Siddique MA, Rashid ESMH, Khalequzzaman M, Bashar MK, Khan LR. Molecular characterization and genetic diversity in T. Aman landraces of rice (Oryza sativa L.) Using microsatellite markers. Thai Journal of Agricultural Science. 2014; 47(4): 211–220. ISSN: 00493589",
"DOI": null,
"article": 193
},
{
"id": 6518,
"serial_number": 27,
"pmc": null,
"reference": "Thomson MJ, Zhao K, Wright M, McNally KL, Rey J, Tung C, et al. High-throughput single nucleotide polymorphism genotyping for breeding applications in rice using the BeadXpress platform. Molecular Breeding. 2012; 29(4): 875–886. DOI: 10.1007/s11032-011-9663-x",
"DOI": null,
"article": 193
},
{
"id": 6519,
"serial_number": 28,
"pmc": null,
"reference": "Somaratne LHMYK, Abayawickrama ASMT, Wickramasinghe IP, Samarasinghe WLG. Estimating Out-Crossing Rate of Bg 379-2 Using Morphological Markers and Confirmation by Molecular Markers. Rice Science. 2012; 19(2): 166–168. DOI: 10.1016/S1672-6308(12)60036-5",
"DOI": null,
"article": 193
},
{
"id": 6520,
"serial_number": 29,
"pmc": null,
"reference": "McCouch SR, Chen X, Panaud O, Temnykh S, Xu Y, Cho Y, et al. Microsatellite marker development, mapping and applications in rice genetics and breeding. Plant Molecular Biology. 1997; 35(1-2): 89-99. DOI: 10.1007/978-94-011-5794-0_9",
"DOI": null,
"article": 193
},
{
"id": 6521,
"serial_number": 30,
"pmc": null,
"reference": "Chen X, Temnykh S, Xu Y, Cho YG, McCouch SR. Development of a microsatellite framework map providing genome-wide coverage in rice (Oryza sativa L.). Theoretical and Applied Genetics. 1997; 95(4): 553-567. DOI: 10.1007/s001220050596",
"DOI": null,
"article": 193
},
{
"id": 6522,
"serial_number": 31,
"pmc": null,
"reference": "Ramu P, Kassahun B, Senthilvel S, Kumar CA, Jayashree B, Folkertsma RT, et al. Exploiting rice-sorghum synteny for targeted development of EST-SSRs to enrich the sorghum genetic linkage map. Theoretical and Applied Genetics. 2009; 119(7): 1193-1204. DOI: 10.1007/s00122-009-1120-4",
"DOI": null,
"article": 193
},
{
"id": 6523,
"serial_number": 32,
"pmc": null,
"reference": "Parida SK, Dalal V, Singh AK, Singh NK, Mohapatra T. Genic non-coding microsatellites in the rice genome: Characterization, marker design and use in assessing genetic and evolutionary relationships among domesticated groups. BMC Genomics. 2009; 10. DOI: 10.1186/1471-2164-10-140",
"DOI": null,
"article": 193
},
{
"id": 6524,
"serial_number": 33,
"pmc": null,
"reference": "Akagi H, Yokozeki Y, Inagaki A, Fujimura T. Microsatellite DNA markers for rice chromosomes. Theoretical and Applied Genetics. 1996; 93(7): 1071-1077. DOI: 10.1007/BF00230127",
"DOI": null,
"article": 193
},
{
"id": 6525,
"serial_number": 34,
"pmc": null,
"reference": "Panaud O, Chen X, McCouch SR. Development of microsatellite markers and characterization of simple sequence length polymorphism (SSLP) in rice (Oryza sativa L.). Molecular and General Genetics MGG. 1996; 252(5): 597-607. DOI: 10.1007/s004380050267",
"DOI": null,
"article": 193
},
{
"id": 6526,
"serial_number": 35,
"pmc": null,
"reference": "Temnykh S, Park WD, Ayres N, Cartinhour S, Hauck N, Lipovich L, et al. Mapping and genome organization of microsatellite sequences in rice (Oryza sativa L.). Theoretical and Applied Genetics. 2000; 100(5): 697-712Theor Appl Genet. 2000. DOI: 10.1007/s001220051342",
"DOI": null,
"article": 193
},
{
"id": 6527,
"serial_number": 36,
"pmc": null,
"reference": "Temnykh S, DeClerck G, Lukashova A, Lipovich L, Cartinhour S, McCouch S. Computational and experimental analysis of microsatellites in rice (Oryza sativa L.): Frequency, length variation, transposon associations, and genetic marker potential. Genome Research. 2001; 11(8): 1441-1452. DOI: 10.1101/gr.184001",
"DOI": null,
"article": 193
},
{
"id": 6528,
"serial_number": 37,
"pmc": null,
"reference": "Rahman L, Alam MS, Hassan L, Alam MS, Bashar MK, Rahman MS, et al. Rice Varieties of Bangladesh: Morphological and Molecular Characterization. Vol 1. Published by Seed Wing, Ministry of Agriculture, Government of the People’s Republic of Bangladesh; 2008. DOI: 10.13140/RG.2.1.3160.9684",
"DOI": null,
"article": 193
},
{
"id": 6529,
"serial_number": 38,
"pmc": null,
"reference": "Rahman M, Sohag M, Rahman L. Microsatellite based DNA fingerprinting of 28 local rice (Oryza sativa L.) varieties of Bangladesh. J Bangladesh Agric Univ. 2010; 8(1): 7-17. DOI: 10.3329/jbau.v8i1.6391",
"DOI": null,
"article": 193
},
{
"id": 6530,
"serial_number": 39,
"pmc": null,
"reference": "Aljanabi S. Universal and rapid salt-extraction of high quality genomic DNA for PCR- based techniques. Nucleic Acids Res. 1997; 25(22): 4692-4693. DOI: 10.1093/nar/25.22.4692",
"DOI": null,
"article": 193
},
{
"id": 6531,
"serial_number": 40,
"pmc": null,
"reference": "Nash J.H.E. DNAfrag, Version 3.03. Institute for biological sciences, National Research Council of Canada, Ottawa, Ontario, Canada. 1991.",
"DOI": null,
"article": 193
},
{
"id": 6532,
"serial_number": 41,
"pmc": null,
"reference": "Nash JHE, Villegas A, Kropinski AM, Aguilar-Valenzuela R, Konczy P, Mascarenhas M, et al. Genome sequence of adherent-invasive Escherichia coli and comparative genomic analysis with other E. coli pathotypes. BMC Genomics. 2010; 11(1): 667. DOI: 10.1186/1471-2164-11-667",
"DOI": null,
"article": 193
},
{
"id": 6533,
"serial_number": 42,
"pmc": null,
"reference": "Yeh FC, Boyle J. POPGENE, the user-friendly softare for population genetic analysis. Molecular Biology and Biotechnology. 1997; 434: 724–31. DOI: 10.1038/nature03466",
"DOI": null,
"article": 193
},
{
"id": 6534,
"serial_number": 43,
"pmc": null,
"reference": "Kimura M, Crow JF. The numer of alleles that can be maintained. Genetics. 1964; 49(4): 725–738. ISSN: 00166731",
"DOI": null,
"article": 193
},
{
"id": 6535,
"serial_number": 44,
"pmc": null,
"reference": "Lewontin RC. The apportionment of human diversity. In: The Concept of Race in Natural and Social Science. 2014; 7-15. Taylor and Francis. DOI: 10.1007/978-1-4684-9063-3_14",
"DOI": null,
"article": 193
},
{
"id": 6536,
"serial_number": 45,
"pmc": null,
"reference": "Levene H. On a Matching Problem Arising in Genetics. The Annals of Mathematical Statistics.1949; 20(1): 91–94. DOI: 10.1214/aoms/1177730093",
"DOI": null,
"article": 193
},
{
"id": 6537,
"serial_number": 46,
"pmc": null,
"reference": "Nei M. Genetic Distance. In: Brenner’s Encyclopedia of Genetics: Second Edition. Elsevier Inc., 2013: 248-250. DOI: 10.1016/B978-0-12-374984-0.00615-X",
"DOI": null,
"article": 193
},
{
"id": 6538,
"serial_number": 47,
"pmc": null,
"reference": "Nei M. F‐statistics and analysis of gene diversity in subdivided populations. Annals of Human Genetics. 1977; 41(2): 225–233. DOI: 10.1111/j.1469-1809.1977.tb01918.x",
"DOI": null,
"article": 193
},
{
"id": 6539,
"serial_number": 48,
"pmc": null,
"reference": "Nei M. Analysis of Gene Diversity in Subdivided Populations Masatoshi Nei Center for Demographic and Population. Proc Nat Acad Sci USA. 1973; 41(2): 225–233. DOI: 10.1111/j.1469-1809.1977.tb01918.x",
"DOI": null,
"article": 193
},
{
"id": 6540,
"serial_number": 49,
"pmc": null,
"reference": "Evening M. Adobe Photoshop CS6 for Photographers. 2012. DOI: 10.4324/9780240526126",
"DOI": null,
"article": 193
},
{
"id": 6541,
"serial_number": 50,
"pmc": null,
"reference": "Nei M. Genetic Distance between Populations. The American Naturalist. 1972; 106(949): 283–292. DOI: 10.1086/282771",
"DOI": null,
"article": 193
},
{
"id": 6542,
"serial_number": 51,
"pmc": null,
"reference": "Page RDM. Treeview: An application to display phylogenetic trees on personal computers. Bioinformatics. 1996;12(4):357-358. DOI: 10.1093/bioinformatics/12.4.357",
"DOI": null,
"article": 193
},
{
"id": 6543,
"serial_number": 52,
"pmc": null,
"reference": "Yu JK, Dake TM, Singh S, Benscher D, Li W, Gill B, et al. Development and mapping of EST-derived simple sequence repeat markers for hexaploid wheat. Genome. 2004; 47(5): 805–818. DOI: 10.1139/G04-057",
"DOI": null,
"article": 193
},
{
"id": 6544,
"serial_number": 53,
"pmc": null,
"reference": "Yang GP, Saghai Maroof MA, Xu CG, Zhang Q, Biyashev RM. Comparative analysis of microsatellite DNA polymorphism in landraces and cultivars of rice. MGG Molecular & General Genetics. 1994; 245(2): 187-194. DOI: 10.1007/BF00283266",
"DOI": null,
"article": 193
},
{
"id": 6545,
"serial_number": 54,
"pmc": null,
"reference": "Khalequzzaman M, Islam MZ, Siddique MA, Prince MFRK, Rashid ESMH, Ahamed MS. Genetic Diversity in AUS Rice Landraces of Bangladesh using SSR Markers. Bangladesh Journal of Plant Breeding and Genetics. 2017; 30(1); 11–20. DOI: 10.3329/bjpbg.v30i1.36529",
"DOI": null,
"article": 193
},
{
"id": 6546,
"serial_number": 55,
"pmc": null,
"reference": "Debouck DG. Managing Plant Genetic Diversity. Crop Science. 2003; 43(2): 749-750. DOI:10.2135/cropsci2003.749a",
"DOI": null,
"article": 193
},
{
"id": 6547,
"serial_number": 56,
"pmc": null,
"reference": "Ni J, Colowit PM, Mackill DJ. Evaluation of genetic diversity in rice subspecies using microsatellite markers. Crop Science. 2002; 42(2): 601-607. DOI: 10.2135/cropsci2002.6010",
"DOI": null,
"article": 193
},
{
"id": 6548,
"serial_number": 57,
"pmc": null,
"reference": "Sharmin A, Hoque ME, Haque MM, Khatun F. Molecular Diversity Analysis of Some Chilli (Capsicum spp.) Genotypes Using SSR Markers. American Journal of Plant Sciences. 2018; 9(03): 368-379. DOI: 10.4236/ajps.2018.93029",
"DOI": null,
"article": 193
},
{
"id": 6549,
"serial_number": 58,
"pmc": null,
"reference": "Cao T, Duprez E, Borden KLB, Freemont PS, Etkin LD. Ret finger protein is a normal component of PML nuclear bodies and interacts directly with PML. Journal of Cell Science. 1998; 111(10): 1319–1329. ISSN: 00219533",
"DOI": null,
"article": 193
},
{
"id": 6550,
"serial_number": 59,
"pmc": null,
"reference": "Mia MM, Rahman S, Islam MM, Begum SN, Hassan L. Molecular characterization of rice genotypes for Zinc biosynthetic gene(s) using microsatellite simple sequence repeat (SSR) markers. Asian Journal of Medical and Biological Research. 2015; 1(2): 187–197. DOI: 10.3329/ajmbr.v1i2.25611",
"DOI": null,
"article": 193
},
{
"id": 6551,
"serial_number": 60,
"pmc": null,
"reference": "Hossain MM, Islam MM, Hossain H, Jaime A, Silva TD, Komamine A, et al. Genetic Diversity Analysis of Aromatic Landraces of Rice (Oryza sativa L.) by Microsatellite Markers. Genes, Genomes and Genomics. 2012; 6(1): 42-47. Retrieved from http://www.globalsciencebooks.info/Online/GSBOnline/images/2012/GGG_6(SI1)/GGG_6(SI1)42-47o.pdf",
"DOI": null,
"article": 193
},
{
"id": 6552,
"serial_number": 61,
"pmc": null,
"reference": "Gao LZ. Microsatellite variation within and among populations of Oryza officinalis (Poaceae), an endangered wild rice from China. Molecular Ecology. 2005; 14(14): 4287-4297. DOI: 10.1111/j.1365-294X.2005.02758.x",
"DOI": null,
"article": 193
},
{
"id": 6553,
"serial_number": 62,
"pmc": null,
"reference": "Sharma R, Kumar B, Arora R, Ahlawat S, Mishra AK, Tantia MS. Genetic diversity estimates point to immediate efforts for conserving the endangered Tibetan sheep of India. Meta Gene. 2016; 8: 14-20. DOI: 10.1016/j.mgene.2016.01.002",
"DOI": null,
"article": 193
},
{
"id": 6554,
"serial_number": 63,
"pmc": null,
"reference": "Warzecha J, Oczkowicz M, Rubis D, Fornal A, Szmatoła T, Bugno-Poniewierska M. An evaluation of the genetic structure of geese maintained in Poland on the basis of microsatellite markers. Animals. 2019; 9(10): 737. DOI: 10.3390/ani9100737",
"DOI": null,
"article": 193
},
{
"id": 6555,
"serial_number": 64,
"pmc": null,
"reference": "Rahman L, Islam MN , Rahman MS, Islam MS, Alam MS and Bashar MK. Characterization of 94 Rice (Oryza sativa L.) varieties of Bangladesh Based on Microsatellite Loci. Bangladesh Journal of Agricultural Science. 2008; 35(1): 97-112.",
"DOI": null,
"article": 193
}
]
},
{
"id": 196,
"slug": "178-1582884845-habit-and-habitual-status-with-relative-diversity-study-of-avifauna-of-jaipurhat-district-of-bangladesh",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1582884845",
"recieved": "2020-03-27",
"revised": null,
"accepted": "2020-05-29",
"published": "2020-06-13",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/42/178-1582884845.pdf",
"title": "Habit and habitual status with relative diversity study of avifauna of Jaipurhat district of Bangladesh",
"abstract": "<p>This investigation was carried out to observe the migration status and the feeding guild of the avian fauna including the relative diversity of Jaipurhat district of Bangladesh. Data were collected from November 2017 to October 2019. During the study period, 89 avian species were recorded that belong to 32 families under 11 orders among which 8 (8.99%) species were common winter visitors, 3 (3.37%) were common summer visitors, 8 (8.99%) were resident migratory, 1 (1.12%) were rare resident and 69 (77.53%) were resident birds. Among the birds observed in the study area during the study period the highest number of birds was found to be insectivorous (41.57%), then the carnivorous (8.99%), then omnivorous, piscivorous and picio carnivorous (7.87%), then insecto frugivorous (6.67%), then frugivorous (5.62%), then frugio graminivorous (4.49%), then nectarivorous (3.39%), then graminivorous and molluscio carnivorous (2.25%) and insecto carnivorous (1.12%). The hight avian diversity (69.662%) was observed in Mithapur which is human inhabited area with crop lands and a lot of fruit trees and the lowest (32.584%) diversity was in Shree Rampur Bill and Kastogari Bill which has large water bodies. This study will be helpful to understand the status of birds in different season in the study area.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 204-215.",
"academic_editor": "Dr. Dinh-Toi Chu, Hanoi National University of Education, Vietnam.",
"cite_info": "Amin A, Hasan K, et al. Habit and habitual status with relative diversity study of avifauna of Jaipurhat district of Bangladesh. J Adv Biotechnol Exp Ther. 2020; 3(3): 204-215.",
"keywords": [
"birds",
"Migration status",
"Species diversity",
"Relative diversity",
"Feeding guild"
],
"DOI": "10.5455/jabet.2020.d126",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>Zoo geographically Bangladesh is located at the Indo- Malayan and Indo- China sub region of the Oriental region and for this bio geographical location Bangladesh acts as a corridor of wildlife among the neighboring countries, India, Myanmar and Bhutan. Bangladesh is famous for its avian diversity. This country shelters a lot of resident birds and a lot of migratory birds come to this country both in summer and winter for food and shelter. Birds are common dwellers of any ecosystem and very important ecological indicator as well. Diversity of avifauna is one of the most important ecological indicators to evaluate the quality of habitats [<a href=\"#r-1\">1</a>]. In any type of ecosystem, avian fauna constitutes one of the major biotic components [<a href=\"#r-2\">2</a>]. Birds play significant ecological roles like pollination, seed dispersion, maintaining ecological balance etc. The rich avian diversity indicates a healthy ecosystem.<br />\r\nA recent census of birds, informed that being a biodiversity hotspot Bangladesh harbors 690 species of birds [<a href=\"#r-3\">3</a>], which is 7% of the world total species of 9,600 (MFA compiled, unpublished report). Published regional avifauna of Bangladesh are found in literature viz., Islam [<a href=\"#r-4\">4</a>] from Rangpur District; Husain [<a href=\"#r-5\">5, 6</a>] from Chittagong Hill Tracts and Pablakhali Wildlife Sanctuary; Banerjee [<a href=\"#r-7\">7</a>] and Das [<a href=\"#r-8\">8</a>] from Curzan Hall area of the University of Dhaka and the Ramna Park of Dhaka respectively; Das [<a href=\"#r-8\">8</a>] from Sylhet District; Sarker & Sarker [<a href=\"#r-9\">9</a>] from the Sundarbans, Haque & Rahman [<a href=\"#r-10\">10</a>] from the Ramna Park and Suhrawardy Uddyan, Dhaka; Akhter [<a href=\"#r-11\">11</a>] from the Boldha Garden, Dhaka; Sarker et al. [<a href=\"#r-12\">12</a>] from the Moheshkhali Island; Reza et al. [<a href=\"#r-13\">13</a>] from Rajshahi University Campus; and Reza et al. [<a href=\"#r-14\">14</a>] from the Padma Charlands near Rajshahi City, etc. Sarkar [<a href=\"#r-15\">15</a>] published seabirds of the Bay of Bengal of Bangladesh Coast and their conservation; Jaman et al. [<a href=\"#r-16\">16, 17</a>] studied ecology, conservation problems and status of avifauna of Noakhali Charbata Coastal area. Sarker <em>et al</em>. [<a href=\"#r-18\">18, 19</a>] reported diversity of avifauna of Bagkhali Range and Cox’s Bazar; Sarker <em>et al</em>. [<a href=\"#r-20\">20</a>] reported the avifauna from two urban sites of the Uttara Model Town, Dhaka. Literature on the avian diversity are very scanty from the Rajshahi region. Haque [<a href=\"#r-21\">21</a>] published a list of birds from the Rajshahi University campus, which was later updated by Reza et al. [<a href=\"#r-13\">13</a>]. Karmakar <em>et al</em>. [<a href=\"#r-22\">22</a>] published an article on birds of Joypurhat District. Reza et al. [<a href=\"#r-14\">14</a>], Reza & Parween [<a href=\"#r-23\">23</a>] reported on the waterbirds of the Padma River and the migratory birds of the Padma River Charlands, Hasan <em>et al</em>. [<a href=\"#r-24\">24</a>] published an article on birds of Shoipara Beel of Mohanpur Upazilla, Rajshahi District and Amin & Hasan reported on the birds of Naogaon District respectively [<a href=\"#r-25\">25</a>]. This study will illustrate the status of birds of the study area.</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Study area</strong><br />\r\nJoypurhat is a District of Rajshahi Division that has an area of 965.44 sq. km. It is bounded by Dinajpur in the north, Naogaon in the south, and Bogra and Gaibandha in the east, Bogra and Naogaon and West Bengal, India in the west [<a href=\"#r-33\">33</a>]. For the convenience of the field work, the study area was divided into seven sites follows: Chapadal, Shree Rampur Bill, Mithapur, Paharpur, Jogodishpur, Kastogaree bill and Asranga (site A, B, C, D, E, F and G). The study area might be differentiated into two major groups.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Bill areas</strong><br />\r\nShree Rampur Bill (site B) and Kastogari Bill (site F) has large water body with thin human houses and wide agricultural lands.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Human locality</strong><br />\r\nChapadal (site A), Mithapur (site C), Paharpur (site D), Jogodishpur (site E) and Asranga (site G) were human inhabited areas with villages, small ponds, lots of fruit trees and agricultural lands.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Study period</strong><br />\r\nThe study was carried out from November 2017 to October 2019.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Study design</strong><br />\r\nThe study was carried out whole the year round. Specially, during three seasons i.e. Rainy (July to October), winter (November to February) and summer (March to June) seasons, birds were observed by regular visits (At least 8 days in a month). Look and see method was opted for bird’s survey.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Data collection techniques</strong><br />\r\nField data of birds were observed during winter season at morning hours between 6.30 to 9.00 a.m. and in the evening from 4.00 to 6.00 p.m., during summer season at morning hours between 5.00 to 8.00 a.m. and evening from 5.00 to 7.00 p.m. while, during the rainy season at morning hours between 6.00 to 8.30 a.m. and evening from 4.30 to 6.30 p.m. All types’ of habitats were surveyed carefully for birds. The area was visited by walking; sometimes small boat was used to cover the wetland areas. The birds were observed with binoculars (Bushnell 20 × 200 mm with multicoated lens) and photographed using digital cameras (Nikon P 530).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Identification of birds</strong><br />\r\nThe birds were identified from their photographs in the Central library, Seminar library and with the help of the book Name of Bangladesh birds: prespective written by Professor Dr. Aminuzzaman Saleh Reza, Department of Zoology, University of Rajshahi. Also consulted literatures for bird identification were Ali and Ripley [<a href=\"#r-26\">26</a>], Ali [<a href=\"#r-27\">27</a>], Khan [<a href=\"#r-3\">3</a>, <a href=\"#r-28\">28</a>], Halder [<a href=\"#r-29\">29</a>], Grewal [<a href=\"#r-30\">30</a>].</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Migration status</strong><br />\r\nThe birds were categorized according to their status at the study area, which are Common Resident (R), Rare Resident (RR), Resident Migratory (RM), Migratory, summer visitors (MSV) and Migratory, Winter Visitors (MWV).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Social status</strong><br />\r\nThe status of the resting or foraging types were studied. Some birds were found to rest or forage either solitary or in pair or in small parties. The water birds were found to forage mostly in different sized family flocks or in mixed species flocks. The symbols for the social status of the birds are used as S (solitary/single), P (pair), F (flock), LF (large flock), and SF (small flock),</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Feeding guild</strong><br />\r\nFeeding habits of the observed birds were studied. The bird species were identified accordingly their feeding guild. The feeding guilds were categorized as frugivorous (FV), carnivorous (CV), nectivorous (NV), molluscivorous (MV), piscivorous (PV), granivorous (GV), insectivorous (IV) and Omnivorous (OM).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Statistical analysis</strong><br />\r\nSystematic lists were constructed following Ali and Ripley (1996) [<a href=\"#r-26\">26</a>] to yield Relative Diversity (RD):<br />\r\nRelative abundance was assessed as ‘very common’ (seen on 75–100% of visits), ‘common’ (seen on 50–74% of visits), ‘uncommon’ (seen on 25–49% of visits), and ‘rare’ (seen on <25% of visits).</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><strong>Species diversity</strong><br />\r\nA total of 89 species of birds were recorded from the study area. A check list of birds found in the Jaipurhat District is given in <a href=\"#Table-1\">Table 1</a>. Distribution, abundance of birds in different blocks, local status, social status and feeding guild and IUCN status of birds are represented in <a href=\"#Table-2\">Table 2</a>. Number and percentages of families and species under each order and Relative abundance of bird species under each family is given in <a href=\"#Table-3\">Table 3</a>. Among the total 89 species only one (<em>Ichthyophaga ichthyaetus</em>) is globally threatened [<a href=\"#r-31\">31</a>]. The observed species are representatives of 11 orders and 32 families.</p>\r\n\r\n<div id=\"Table-1\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1582884845-table1/\">Table-1</a><strong>Table 1.</strong> Checklist of the birds found in Jaipurhat district.</p>\r\n</div>\r\n\r\n<div id=\"Table-2\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1582884845-table2/\">Table-2</a><strong>Table 2. </strong>Distribution, abundance of birds in different blocks, local status, social status and feeding guild and IUCN status.</p>\r\n</div>\r\n\r\n<div id=\"Table-3\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1582884845-table3/\">Table-3</a><strong>Table 3. </strong>Number and percentages of families and species under each Order and Relative abundance of bird species under each family.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Migration status</strong><br />\r\nAmong the 89 species of birds 8 (8.99%) species were common winter visitors, 3 (3.37%) were common summer visitors, 8 (8.99%) were resident migratory, 1 (1.12%) were rare resident and 69 (77.53%) were resident birds (<a href=\"#Table-2\">Table 2,</a> <a href=\"#figure1\">Figure 1</a>). Among the birds listed in this area, the most common winter visitors were Fulvous whistling duck (<em>Dendrocygna bicolor</em>), Lesser whistling-duck (<em>Dendrocygna javanica</em>), Common Snipe (<em>Gallinago gallinago</em>), Common Swallow (<em>Hirundo rustica</em>), White-Browed Wagtail (<em>Motacilla maderaspatensis</em>), White Wagtail (<em>Motacilla alba</em>), Yellow Wagtail (<em>Motacilla flava</em>) and Tickell’s Leaf wabler (<em>Dicaeum cruenatum</em>); summer visitors were Blue-tailed Bea-Eater (<em>Merops philippinus</em>), Pied Cuckoo (<em>Clamator jacobinus)</em> and Common Hawk Cuckoo (<em>Hierococcyx varius</em>). Grey-Headed fish eagle (<em>Ichthyophaga ichthyaetus</em>) was seen in this study area as well which is considered as a rear resident bird that is a globally threatened at present time according to IUCN. Night Heron (<em>Nycticorax nycticorax</em>), Great Egret (<em>Casmerodicus albus</em>), Asian openbill (<em>Anastomus oscitans</em>), Little Cormorant (<em>Phalacrocorsx carbo</em>), Jungle Crow (<em>Corvus macrorhynchos</em>) and Ashy Drongo (<em>Dicrurus leucophaeus</em>) were seen periodically because of their resident migratory habit. The rest were seen all years around.</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"296\" src=\"/media/article_images/2024/04/15/178-1582884845-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1. </strong>Migration status of birds in the study area.</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Social status</strong><br />\r\nBirds are different in social status as well as their feeding status. Sometime they are found in solitary or in pairs. Some birds are formed small to large groups. Some birds are visited in a small to large or loose flocks. The number of species found as single (solitary) was 13 (14.61%), paired 1 (1.12%) and may single or in pairs was 32 (35.96%). However, 4 (4.49%) species were found in small flock and 2 (2.25%) were found in large flock. Some birds were found in single or small flocks, some were found in paired or small flock while some were seen in small or large flocks and the numbers of these groups of birds are 11 (12.36%), 6 (6.74%) and 4 (4.49%) respectively. About 9 (10.11%) birds were seen in single, paired or forming small flocks; 5 (5.62%) were single, paired or in large flocks, and 2 (2.25%) bird species were seen single or formed both small and large flocks. All these statuses are shown in <a href=\"#Table-2\">Table 2</a> and <a href=\"#figure2\">Figure 2</a>.</p>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"295\" src=\"/media/article_images/2024/04/15/178-1582884845-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2.</strong> Social status of birds in the study area.(S = Single, P = Paired, SF = Small flock, LF = Large flock, S/P = Single or paired, SF/LF = Small or large flock, S/SF = Single or small flock, P/SF = Pared or small flock, S/P/SF = Single or paired or small flock, S/P/LF = Single or paired or large flock, S/SF/LF = Single or Small flock or large flock.Caption</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Feeding guild</strong><br />\r\nThe guild-wise percentages of the 89 species are shown in <a href=\"#figure3\">Figure 3</a>. Among the species 41.57% were insectivorous, followed by omnivorous 7.87%, carnivorous 8.99%, fruityvorous 5.62%, piscivorous 7.87%, nectarivorous 3.37% and graminivorous 2.25%. Some species has also intermediate feeding guild such as picivorous-carnivorous 7.87% and insectivorous-fruityvorous 6.74%, frugivorous-graminivorous 4.49%, insectivorous-carnivorous 1.12% and molluscivorous-carnivorous 2.25%.</p>\r\n\r\n<div id=\"figure3\">\r\n<figure class=\"image\"><img alt=\"\" height=\"296\" src=\"/media/article_images/2024/04/15/178-1582884845-Figure3.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 3.</strong> Feeding guild of birds of in study area (OM= Omnivorous, CV = Carnivorous, IV = Insectivorous, FV = Frugivorous, PV = Piscivorous, GV = Graminivorous, NV = Nectarivorous, IV+FV = insecto frugivorous, FV + GV frugio graminivorous, PV + CV = picio carnivorous, IV + CV = insecto carnivorous, MV + CV = molluscio carnivorous).</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Relative diversity of birds on the basis of different study sites</strong><br />\r\nBird diversity and dispersion depends on the habitat and availability of the food sources. For example aquatic birds like duck, cormorant etc. are very common near water bodies, fruit eaters are seen in trees usually, and grain eaters love to forage in the crop fields. The study area was divided into seven sites under two major habitat, bill area and human locality. The relative diversity of birds on the basis of different study sites is given in <a href=\"#figure4\">figure 4</a>. Bird diversity was found higher in human locality than the bill areas. In Mithapur the highest bird diversity was observed (69.622%), then in Chapadal (66.292%), then in Jogodishpur (65.168%), then in Asranga (59.55%). The lowest diversity was observed in Shree Rampur Bill and Kastogaree Bill (32.584%)</p>\r\n\r\n<div id=\"figure4\">\r\n<figure class=\"image\"><img alt=\"\" height=\"296\" src=\"/media/article_images/2024/04/15/178-1582884845-Figure4.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 4. </strong>Relative diversity of birds.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>During this study period 89 bird species were observed that are the representatives of 11 orders and 32 families which support the findings of Karmakar <em>et al</em>. [<a href=\"#r-22\">22</a>]. According to Siddiqui <em>et al</em> [<a href=\"#r-32\">32</a>] there are a total 650 species of birds found in the Bangladesh territory, among which 477 are regular species. Out of these regular species 301 are residents of the country and 176 are seasonal migrants. The number of winter migrants is 160, summer migrants are 6 and spacing or passage migrants are 10. Based on relative abundance of birds 143 are vagrants, 176 are rare, 103 are uncommon and 198 are common species of Bangladesh [<a href=\"#r-3\">3</a>, <a href=\"#r-28\">28 </a>and <a href=\"#r-32\">32</a>]. Among the birds of Bangladesh IUCN declared 40 species as globally threatened. But Rhett A. Butler mentioned only 607 birds are present and 36 are threatened in Bangladesh in ‘‘Total Number of Bird Species by Country’’ at July 1, 2019 [<a href=\"#r-35\">35</a>]. The avifauna of Jaipurhat District enlists total 89 species which is 13.69% of the Bangladeshi species. The study area includes Small River, marsh land, wetland, cropland, village groves and orchards along with some fallow lands. During this study birds have been recorded 8.99% common winter visitors, 3.37% common summer visitors, 8.99% resident migratory, 1.12% were rare resident and 77.53% resident birds among the total amount of birds.<br />\r\nIn this study according to social status the findings was that the highest number of bird species as single or in pairs 35.96% ; others followed by 14.61% solitary, 1.12% paired, small flock 4.49%, large flock 2.25% , single or small flocks 12.36%, paired or small flock 6.74%, small or large flocks 4.49% respectively. About 10.11% birds were seen in single, paired or forming small flocks; 5.62% were single, paired or in large flocks, and 2.25% bird species were seen single or formed both small and large flocks. And according to feeding guild the highest number of bird species as insectivorous 41.57% and lowest number of bird species as insectivorous-carnivorous 1.12% were recorded. Bird diversity was found higher in human locality than the bill areas. In Mithapur the highest bird diversity was observed (69.622%), then in Chapadal (66.292%), then in Jogodishpur (65.168%), then in Asranga (59.55%). The lowest diversity was observed in Shree Rampur Bill and Kastogaree Bill (32.584%).</p>"
},
{
"section_number": 5,
"section_title": "ACKNOWLEDGEMENTS",
"body": "<p>The authors are grateful to the Chairman, Department of Zoology, University of Rajshahi for providing necessary facilities to accomplish the research work and to Dr. Nurul Islam and Dr. Aminuzzaman Mohammad Saleh Reza for their guidance. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.</p>"
},
{
"section_number": 6,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>AA and KH were involved in conception and design of the experiments. AA, KH, SM, MA, SS and SA contributed to perform the experiments. AA, KH and SM analyzed data. SS contributed to drafting the article. KH and MA contributed to revising it critically for important intellectual content. AA made the final approval of the version to be published.</p>"
},
{
"section_number": 7,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/04/15/178-1582884845-Figure1.jpg",
"caption": "Figure 1. Migration status of birds in the study area.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/04/15/178-1582884845-Figure2.jpg",
"caption": "Figure 2. Social status of birds in the study area.(S = Single, P = Paired, SF = Small flock, LF = Large flock, S/P = Single or paired, SF/LF = Small or large flock, S/SF = Single or small flock, P/SF = Pared or small flock, S/P/SF = Single or paired or small flock, S/P/LF = Single or paired or large flock, S/SF/LF = Single or Small flock or large flock.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/04/15/178-1582884845-Figure3.jpg",
"caption": "Figure 3. Feeding guild of birds of in study area (OM= Omnivorous, CV = Carnivorous, IV = Insectivorous, FV = Frugivorous, PV = Piscivorous, GV = Graminivorous, NV = Nectarivorous, IV+FV = insecto frugivorous, FV + GV frugio graminivorous, PV + CV = picio carnivorous, IV + CV = insecto carnivorous, MV + CV = molluscio carnivorous).",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/04/15/178-1582884845-Figure4.jpg",
"caption": "Figure 4. Relative diversity of birds.",
"featured": false
}
],
"authors": [
{
"id": 864,
"affiliation": [
{
"affiliation": "Institute of Biological Sciences, University of Rajshahi, Rajshahi, 6207, Bangladesh"
}
],
"first_name": "Al",
"family_name": "Amin",
"email": "alaminology1@gmail.com",
"author_order": 1,
"ORCID": null,
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Al Amin, Institute of Biological Sciences, University of Rajshahi, Rajshahi, 6207, Bangladesh, E mail: alaminology1@gmail.com",
"article": 196
},
{
"id": 865,
"affiliation": [
{
"affiliation": "Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal"
}
],
"first_name": "Kamrul",
"family_name": "Hasan",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 196
},
{
"id": 866,
"affiliation": [
{
"affiliation": "Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi, 6207, Bangladesh"
}
],
"first_name": "Shafi",
"family_name": "Mahmud",
"email": null,
"author_order": 3,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 196
},
{
"id": 867,
"affiliation": [
{
"affiliation": "Faculty of Fine Arts, University of Rajshahi, Rajshahi, 6207, Bangladesh"
}
],
"first_name": "Maria",
"family_name": "Akter",
"email": null,
"author_order": 4,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 196
},
{
"id": 868,
"affiliation": [
{
"affiliation": "Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal"
}
],
"first_name": "Shabnam",
"family_name": "Sabiha",
"email": null,
"author_order": 5,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 196
},
{
"id": 869,
"affiliation": [
{
"affiliation": "Rajshahi medical College, Dental unit, Rajshahi, Bangladesh"
}
],
"first_name": "Sumaiya",
"family_name": "Arabi",
"email": null,
"author_order": 6,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 196
}
],
"views": 961,
"downloads": 135,
"references": [
{
"id": 6658,
"serial_number": 1,
"pmc": null,
"reference": "Sarkar NJ, Sultana D, Jaman MF, Rahman MK. Diversity and population of Avifauna of Two Urban States in Dhaka, Bangladesh. Ecological society, Nepal. 2009; 16: 1-7.",
"DOI": null,
"article": 196
},
{
"id": 6659,
"serial_number": 2,
"pmc": null,
"reference": "Dhindas MS, Saini HK. Avian Fauna of Wetlands. Unpublished thesis, Department of Zoology, Punjab Agriculture Univesity, Ludhiana, Pakistan. 1994.",
"DOI": null,
"article": 196
},
{
"id": 6660,
"serial_number": 3,
"pmc": null,
"reference": "Khan MAR. Bangladesher Pakhi (Birds of Bangladesh). Bangla Academy, Dhaka, Bangladesh. 2008.",
"DOI": null,
"article": 196
},
{
"id": 6661,
"serial_number": 4,
"pmc": null,
"reference": "Islam MT. Birds of Rangpur (with notes on taxonomy, food and relationships). M.Sc. Thesis (Unpublished). University of Dhaka. 1969.",
"DOI": null,
"article": 196
},
{
"id": 6662,
"serial_number": 5,
"pmc": null,
"reference": "Husain KZ. Field notes on the birds of the Chittagong Hill Tract. J. Asiat. Soc. Pakistan. 1969; 13(1): 91-101.",
"DOI": null,
"article": 196
},
{
"id": 6663,
"serial_number": 6,
"pmc": null,
"reference": "Husain KZ. Birds of the Pablakhali Wildlife Sanctuary. Bangladesh J. Zool. 1975; 3(2): 145-155.",
"DOI": null,
"article": 196
},
{
"id": 6664,
"serial_number": 7,
"pmc": null,
"reference": "Banerjee RK. Status and composition of avifauna of the Curzon Hall Campus and Ramna Park, Dhaka. M. Sc. thesis, Department of Zoology, University of Dhaka, Bangladesh. 1978.",
"DOI": null,
"article": 196
},
{
"id": 6665,
"serial_number": 8,
"pmc": null,
"reference": "Das SR. Birds of Sylhet. M. Sc. thesis, Department of Zoology, University of Dhaka, Bangladesh. 1975.",
"DOI": null,
"article": 196
},
{
"id": 6666,
"serial_number": 9,
"pmc": null,
"reference": "Sarker SU and Sarker, NJ. Status and distribution of birds of the Sundarbans. J. Noami.1986; 3(1): 19-37.",
"DOI": null,
"article": 196
},
{
"id": 6667,
"serial_number": 10,
"pmc": null,
"reference": "Haque NM and Rahman MK. Birds of the Ramna and Suhrawardy Parks. Published jointly by the Wildlife Society of Bangladesh (WSB) and Bangladesh Bird Preservation Society (BBPS). 1993.",
"DOI": null,
"article": 196
},
{
"id": 6668,
"serial_number": 11,
"pmc": null,
"reference": "Akter S. The wildlife of Boldha garden, Dhaka, Bangladesh (with notes on ecology, habitat and present status). M. Sc. thesis, Department of Zoology, University of Dhaka, Bangladesh. 1997.",
"DOI": null,
"article": 196
},
{
"id": 6669,
"serial_number": 12,
"pmc": null,
"reference": "Sarker SU, Jaman MF, Hossain ML and Sarker NJ. Wildlife diversity of Moheskhali Island: Their ecology and conservation issues. J. Noami. 2001; 18(1): 17-31.",
"DOI": null,
"article": 196
},
{
"id": 6670,
"serial_number": 13,
"pmc": null,
"reference": "Reza AMS, Hossain MM, Hasan MA and Parween S. Anotated checklist of birds of Rajshahi University Campus: An update. j. zool. Rajshahi Univ. 2012; 31: 39-47.",
"DOI": null,
"article": 196
},
{
"id": 6671,
"serial_number": 14,
"pmc": null,
"reference": "Reza AMS, Al-Amin M, Rahman SMA, Hasan MS, Islam MS and Parween, S. A checklist of waterbirds of the Padma river charland (Godagari to Charghat), Rajshahi, Bangladesh. Univ. j. zool., Rajshahi Univ. 2014; 33: 19-23.",
"DOI": null,
"article": 196
},
{
"id": 6672,
"serial_number": 15,
"pmc": null,
"reference": "Sarker SU. Seabirds of the Bay of Bengal off Bangladesh coast and their conservation. Tiger Paper. 1984; 11(4): 9-13.",
"DOI": null,
"article": 196
},
{
"id": 6673,
"serial_number": 16,
"pmc": null,
"reference": "Jaman MF and Sahreen N. Ecology and breeding biology of Chestnut-tailed starling, Sturnus malabaricus (Gmellin 1789). Ecoprint. 2004; 11(1): 27-34.",
"DOI": null,
"article": 196
},
{
"id": 6674,
"serial_number": 17,
"pmc": null,
"reference": "Jaman MF, Haque MSZ, and Sarker SU. Ecology, conservation problems and status of avifauna of Noakhali Charbata Coastal area. J. Noam. 2004; 21(1): 1-13.",
"DOI": null,
"article": 196
},
{
"id": 6675,
"serial_number": 18,
"pmc": null,
"reference": "Sarker SU, Jaman MF, Sarker NJ and Hossain ML. Status of wildlife of St. Martin Island, Bangladesh. Dhaka Univ. J. Biol. Sci. 2000; 9(1): 75-85.",
"DOI": null,
"article": 196
},
{
"id": 6676,
"serial_number": 19,
"pmc": null,
"reference": "Sarker SU, Jaman MF, Sarker NJ and Rahman MK. Diversity of avifauna in Bagkhali range, Cox’s Bazar forest division. Bangladesh Environ. 2000; 230-238.",
"DOI": null,
"article": 196
},
{
"id": 6677,
"serial_number": 20,
"pmc": null,
"reference": "Sarkar NJ, Sultana D, Firoj JM and Rahman MK. Diversity and population of avifauna of two urban sites in Dhaka, Bangladesh. Ecoprint. 2009; 16: 1-7.",
"DOI": null,
"article": 196
},
{
"id": 6678,
"serial_number": 21,
"pmc": null,
"reference": "Haque AKMF. Birds of the Rajshahi University Campus. Bangladesh J. Zool. 1976; 4(2): 125127.",
"DOI": null,
"article": 196
},
{
"id": 6679,
"serial_number": 22,
"pmc": null,
"reference": "Karmakar S, Parween S and Reza AMS. Birds of Joypurhat District, Bangladesh. J. Life & Earth Sci. 2011; 6: 51-57.",
"DOI": null,
"article": 196
},
{
"id": 6680,
"serial_number": 23,
"pmc": null,
"reference": "Reza AMS and Parween S. A preliminary list of the migratory birds of the Padma Charland, Rajshahi. Jahangirnagar University J. Biol. Sci. 2014; 3(2): 85-92.",
"DOI": null,
"article": 196
},
{
"id": 6681,
"serial_number": 24,
"pmc": null,
"reference": "Hasan K, Das BC, Parween S and Reza AMS. Avian diversity of the shoipara beel of mohanpur upazilla, rajshahi, bangladesh. J. Life Earth Science. 2017; 12: 21-27.",
"DOI": null,
"article": 196
},
{
"id": 6682,
"serial_number": 25,
"pmc": null,
"reference": "Amin A and Hasan K. A checklist and relative diversity of avifauna of Atrai, Raninagar and Naogan Sadar Upazillas of Naogaon district of Bangladesh. International Journal of Zoology Studies. 2019. 3(2): 38-45.",
"DOI": null,
"article": 196
},
{
"id": 6683,
"serial_number": 26,
"pmc": null,
"reference": "Ali S and Ripley SD. Handbook of the birds of India and Pakistan, 1. Bombay: Oxford University Press. 1996, pp 266.",
"DOI": null,
"article": 196
},
{
"id": 6684,
"serial_number": 27,
"pmc": null,
"reference": "Ali S. The Book of Indian Birds. Oxford University Press. 2002, pp 466.",
"DOI": null,
"article": 196
},
{
"id": 6685,
"serial_number": 28,
"pmc": null,
"reference": "Khan MAR. Wildlife of Bangladesh: Checklist-cum-Guide. Chayabithi, Dhaka, Bangladesh. 2015.",
"DOI": null,
"article": 196
},
{
"id": 6686,
"serial_number": 29,
"pmc": null,
"reference": "Halder RR. A Photographic Guide to Birds of Bangladesh. Baikal Teal Production; 1st 2010, pp 257.",
"DOI": null,
"article": 196
},
{
"id": 6687,
"serial_number": 30,
"pmc": null,
"reference": "Grewal B, Sujoy M and Wright G. Birds of India, Pakistan, Bangladesh, Nepal & Shri Lanka: A photographic guide. Guidebook Company Ltd. New Delhi, India. 1993.",
"DOI": null,
"article": 196
},
{
"id": 6688,
"serial_number": 31,
"pmc": null,
"reference": "IUCN, Red Data Book of Threatened Birds of Bangladesh. The World Conservation Union, Dhaka, Bangladesh. 2000.",
"DOI": null,
"article": 196
},
{
"id": 6689,
"serial_number": 32,
"pmc": null,
"reference": "Siddiqui NA, Shahidullah M and Shahjalal MAH. An overview of diversity, trends and threats of birds. Indian J. Forestry. 2008; 14(2): 17-45.",
"DOI": null,
"article": 196
},
{
"id": 6690,
"serial_number": 33,
"pmc": null,
"reference": "Banglapedia 2003. National Encyclopedia. Asiatic Society of Bangladesh, Dhaka. 9: 470-474.",
"DOI": null,
"article": 196
},
{
"id": 6691,
"serial_number": 34,
"pmc": null,
"reference": "Banglapedia 2015; http://en.banglapedia.org/index.php?title=Bird.",
"DOI": null,
"article": 196
},
{
"id": 6692,
"serial_number": 35,
"pmc": null,
"reference": "Rhett AB. Total number of birds species by country. ©Mongabay 1995-2020. 2019. https://rainforests.mongabay.com/03birds.htm#content.",
"DOI": null,
"article": 196
}
]
},
{
"id": 194,
"slug": "178-1585380993-antibacterial-and-cytotoxic-activities-of-four-selected-summer-season-fruits-seeds-of-bangladesh",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1585380993",
"recieved": "2020-03-15",
"revised": null,
"accepted": "2020-05-25",
"published": "2020-06-13",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/33/178-1585380993.pdf",
"title": "Antibacterial and cytotoxic activities of four selected summer season fruits’ seeds of Bangladesh",
"abstract": "<p>The seeds of fruits which are thrown away as waste can be used in medicinal and therapeutic purposes. The goal of this investigation was to find out antibacterial and cytotoxic potentialities of crude acetone and ethanol extracts of seeds of four common summer fruits, purchased from Khulna region, Bangladesh: <em>Litchi chinensis</em> (Lychee), <em>Phoenix dactylifera</em> (Date Palm), <em>Syzigum cumini</em> (Black Plum) and <em>Artocarpus heterophyllus</em> (Jackfruit). Extracts were screened for their antibacterial activity by agar well diffusion method, followed by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) determination by colorimetric broth macro dilution method and plating method. The highest antibacterial activity was found from acetone extract of <em>S. cumini</em> against <em>Salmonella typhi</em> where zone of inhibition was 19.67±0.27 mm along with MIC 0.0585 mg/ml and MBC 0.2344 mg/ml. Acetone extract of <em>S. cumini</em> was found to have bactericidal effect on all microorganisms except bacteriostatic on <em>E. coli</em> and <em>S. paratyphi</em> and ethanol extract showed bactericidal effect on all the microorganisms tested. The cytotoxic activity was determined by brine shrimp lethality assay. Among all seed samples, <em>P. dactylifera</em> exerted relatively better cytotoxicity where LC<sub>50</sub> value was below 100 µg/ml. The rest of the samples showed relatively less toxicity as LC<sub>50</sub> value was more than 100 µg/ml. Among all the seed samples, <em>S. cumini</em> showed most potential antibacterial activity with less cytotoxicity. Overall, the findings of the study endorse antibacterial activity of examined seed extracts with moderate cytotoxic activity, suggesting the isolation of active compounds thorough bioassays.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 194-203.",
"academic_editor": "Dr. Akhi Moni, ABEx Bio-Research Center, Bangladesh.",
"cite_info": "Rahman SA, Akhter MS. Antibacterial and cytotoxic activities of four selected summer season fruits’ seeds of Bangladesh. J Adv Biotechnol Exp Ther. 2020; 3(3): 194-203.",
"keywords": [
"Cytotoxicity",
"Antibacterial activity",
"Phoenix dactylifera",
"Syzigum cumini",
"Litchi chinensis",
"Artocarpus heterophyllus"
],
"DOI": "10.5455/jabet.2020.d125",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>Natural sources are boundless inspiration for developing novel drugs [<a href=\"#r-1\">1</a>]. Natural products do possess enormous diversity both structurally and chemically which are not similar to the library of artificial synthetic small molecules and encourage unique exploration in biochemical and medical sciences [<a href=\"#r-2\">2</a>]. Plants are the simplest sources of natural products and medicines. Numbers of different biologically potent compounds can be produced by plants. Nearly 20% of plants known have been utilized in studies of pharmaceutics and therapeutics impacting the drug industry in progressive ways such as treating cancer and harmful diseases. Presently, according to the estimation of World Health Organization (WHO), eighty percent of the people worldwide use herbal medicine for different aspects of primary health care [<a href=\"#r-3\">3</a>]. Many pharmaceutical companies are showing interest in ‘Green Medicine’, the drugs driven from plants mainly because of the faith of safety and acceptances compare to synthesized drug, which can possess conflicting effects [<a href=\"#r-4\">4</a>].<br />\r\nAntibiotic drugs which have saved and still saving thousands of lives every year, “antibiotic resistance” could be a serious and one in every of the foremost threatening problems worldwide, including Bangladesh [<a href=\"#r-5\">5</a>]. Antibiotics, the frequently recommended medicines in normal medical practicing but drug resistant diseases remain priority concern in public health issues [<a href=\"#r-6\">6</a>]. Resistance to antibiotics and also the rise within the failure of obtainable chemotherapeutics are now resulting in the sorting out therapeutic floras for potential antibacterial, antiviral, antifungal, antiprotozoal and anticancer function, as well. Failure of available synthetic drugs to treat multiple drug resistant bacteria and undesirable side effects of chemotherapeutics suggest to findout alternative drug sources to overcome the problems [<a href=\"#r-7\">7-8</a>]. Traditional medical practitioners in Bangladesh is an integral part of the culture of using plant or its parts as therapeutic agents, without any dependency on synthetic drugs: to them, plants driven compounds are the first choice.<br />\r\nWith the advancement of medicinal technology, it’s now easier to spot specific botanical constituents and assess their potential activity. Conducting contemporary research dealing with local plant samples characterize the therapeutic values of plant derived constituents and evaluate their prospective potential activity of the available plants of a specific region. Pharmacological prospecting of plants by established method for the identification of effective compounds can lead to the development of novel medicinal agents. Bio prospecting of plants might show qualitative and quantitative variation of effective pharmacological ingredients due to difference in geographical location. In this favor, Bangladesh is a country full of biodiversity. Lots of plants of different regions are available having medicinal properties for the treatment of assorted diseases. However, scientific proofing are conducted to a limited extent with few medicinal plants [<a href=\"#r-9\">9</a>]. About 5,700 species of angiosperms have been recorded from this country. But only about 500 species have documented scientifically as medicinal plants [<a href=\"#r-10\">10</a>].<br />\r\nOut of all the summer season fruits of Bangladesh, the designated four fruits in this study were selected as they are worth mentioning due to their belongingness to the popular crowd favorites. Interestingly all these four fruits are grown and accordingly found in the market of Bangladesh for comparatively shorter period of time than the rest of the summer season fruits of Bangladesh. On the other hand, normally plant parts: leaf, bark, root, fruits, flowers are extensively tested <em>in vitro</em> and <em>in vivo</em> for their pharmacological profiling. Seeds of plants, more specifically, of ripe fruits that are thrown away as wastes can possess pharmacological activities, too. So, taken together all these into consideration, in this study, seeds of four summer season fruits: <em>S. cumini,</em> <em>P. dactylifera</em>, <em>L. chinensis</em> and <em>A. heterophyllus </em>of Bangladesh have been tested <em>in vitro</em> to evaluate preliminary antibacterial and cytotoxic activities.</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Seed collection</strong><br />\r\nFresh ripe fruits were purchased from local market of Khulna district of Bangladesh. Fruits were cut into pieces to extract out seeds from ripe fruits. After bringing out seeds of fruits, they were washed, and sun dried with shade to avoid dust.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Sample preparation and extraction</strong><br />\r\nSeeds were grinded into fine powder. One hundred fifty gm of the powder were soaked into 500 ml of absolute acetone and ethanol solvents and kept in dark places for ten days with occasional shaking, followed by, filtration through Whatman filter paper, evaporation using rotary evaporator and air drying at room temperature. Amount and final color of the extracts were as illustrated in <a href=\"#Table-1\">Table 1</a>.</p>\r\n\r\n<div id=\"Table-1\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1585380993-table1/\">Table1</a><strong>Table 1. </strong>Amount and color of extract obtained after ten days of soaking.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Antibacterial screening</strong><br />\r\n<em>Agar well diffusion assay</em><br />\r\nBacteria were grown on Nutrient Agar Medium (yeast extract 2.0g, beef extract 1.0g, NaCl 5.0g, peptone 5.0g, agar 15g, and distilled water 1 L). Dilution with 10<sup>5</sup> CFU/ml was used as inoculum for the study. Molted agar medium was poured into petri dishes to obtain a depth of 10 mm. The inoculation of the entire surface area of the agar plate was done with 500 µl inoculum. Then, with the help of a disinfected cork borer four holes of 5 mm span were made by punching in aseptic condition. Fifty and hundred µl of sample was introduced into two holes: one kept blank and in another 5 µl of gentamycin (200 µg/ml) was added as positive control. For agar well diffusion test, sample was prepared at a concentration of 20 mg extract into 1 ml of acetone or ethanol. Agar plates were incubated, and inhibition zone was measured as per Balouiri et al., 2016 [<a href=\"#r-11\">11</a>]. The following bacteria were used: Escherichia coli (American Type Culture Collection, ATCC-8739), Pseudomonas aeruginosa (ATCC-27833), Vibrio cholerae (ATCC-14035), Salmonella typhi (ATCC-14028), Salmonella paratyphi (ATCC-13311), Micrococcus spp. (ATCC-14396), and Staphylococcus aureus (ATCC-25923). The findings of the agar well diffusion assay were expressed in four categories depending on zone (diameter) of inhibition: designated as ‘not active’, ‘partially active’, ‘active’, and ‘very active’ when zone (diameter) of inhibition were <9 mm, 9 – 12 mm, 13 – 18 mm, >18 mm, respectfully [<a href=\"#r-12\">12</a>].</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>MIC assay</em><br />\r\nThe assay was performed following a modified method [<a href=\"#r-13\">13</a>]: a mannitol phenol red broth was used. A stock solution of samples were prepared: Nine consecutive two fold dilutions of extract with 50% Dimethyl Sulfoxide to obtain stock concentrations of 50 mg/ml to 0.195 mg/ml. One-tenth ml of stock sample from previously prepared was mixed into 0.4 ml broth. For nine different concentrations, nine different tubes were used respectively to obtain final concentrations of 10 mg/ml to 0.039 mg/ml. Now into each test tube, 10µl of freshly prepared bacterial culture was nurtured at 37ºC overnight. As positive control, Gentamycin was used at a concentration from 10 to 0.039 µg/ml. As negative control, test tubes without sample but with bacterial culture were used. MIC was tested for those microorganisms and sample for which zone of inhibition was 13 mm or above.</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>MBC assay</em><br />\r\nThis method is required to identify the concentration at which 100% microorganisms will be killed. 10 µl of broth from test tubes from where MIC was considered were plated in nutrient agar plate and incubated to see the growth.</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>MIC (MBC/MIC) indexing</em><br />\r\nThe MIC index was recorded to find out whether an extract is bactericidal (MBC/MIC <4) or not. The range of MIC index value greater than 4 but less than 32 were considered as bacteriostatic [<a href=\"#r-8\">8</a>].</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Cytotoxic activity testing</strong><br />\r\nTo ascertain the cytotoxic activity of an extract, the brine shrimp lethality test is acknowledged as standard [<a href=\"#r-14\">14</a>]. For the experiment, 5mg of each extract were dissolved in 1ml of seawater and one drop Tween-80; adjusted to a final concentration of 5μg/μl. Then 4ml of seawater was given to each of the test tubes. From the stock solution specific volumes of 5, 10, 20, 40, 80, 160 and 320 μl of samples were poured into the test tubes; adjusted to 10 ml with saline water to get the final concentration of 2.5, 5, 10, 20, 40, 80, 160 μg/ml, respectively. Finally, 10 live brine shrimp nauplii were taken into each test tube [<a href=\"#r-14\">14</a>]. As positive control, vincristine sulfate was used. Following the mortality count, after 24 hours; log concentration versus percent mortality were plotted and median lethal concentration (LC<sub>50</sub>) were calculated.<br />\r\nOf note, the findings of the agar well diffusion assay were studied in Microsoft Excel 2013. In cytotoxic activity testing, LC<sub>50</sub> values were also calculated by using Microsoft Excel 2013. Each test was conducted three times.</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><strong>Effect on antibacterial activity</strong><br />\r\nThe zone of inhibition for 100 µl/well is higher, as expected, than 50 µl/well (<a href=\"#figure1\">Figure 1</a> and <a href=\"#figure2\">2</a>). The topmost finding was obtained from<em> S. cumini </em>acetone extract against <em>S. typhi</em> having zone of inhibition for 100µl/well as 19.67±0.27 mm (<a href=\"#figure1\">Figure 1</a>) with MIC and MBC value 0.0585mg/ml and 0.2344 mg/ml respectively (<a href=\"#Table-2\">Table 2</a>, <a href=\"#Table-3\">3</a>, <a href=\"#Table-4\">4</a> and <a href=\"#Table-5\">5</a>; <a href=\"#figure1\">Figure 1</a>, <a href=\"#figure2\">2</a> and <a href=\"#figure3\">3</a>). But from MIC index we found that acetone extract of<em> S. cumini</em> was found to have bactericidal effect on all microorganisms except bacteriostatic on <em>E. coli </em>and <em>S.paratyphi </em>and ethanol extract showed bactericidal effect on all the microorganisms tested (<a href=\"#Table-5\">Table 5</a>). Acetone extract of <em>P. dactylifera </em>showed highest zone 16.67±0.27 mm against S. aureus with MIC and MBC value of 1.25 and 5 mg/ml respectively. Zone of inhibition of <em>P. dactylifera</em> acetone extract was in between 13 – 18 mm for all the tests except <em>S. paratyphi</em>. Again, ethanol extract of P. dactylifera showed highest zone 15.67±0.27mm against <em>P. aeruginosa</em> with MIC and MBC value of 1.25 and 7.5 mg/ml, respectively.<br />\r\nAcetone extract of <em>L. chinensis</em> was found active against all the microorganisms tested and found very active against S. aureus (<a href=\"#Table-2\">Table 2</a>, <a href=\"#Table-3\">3</a> and <a href=\"#Table-4\">4</a>). Ethanol extract of <em>L. chinensis</em> was found active against <em>P. aeruginosa</em>, <em>Micrococcus</em> spp., <em>E. coli</em> and <em>S. aureus, </em>and partially active against rest of the tested bacteria. Acetone extract of <em>A. heterophyllus </em>was found partially active against all the tested bacteria except <em>P. aeruginosa</em> (14.67±0.98 mm) and <em>Micrococcus</em> spp. (14.67±0.27 mm). Ethanol extract of <em>A. heterophyllus </em>was found partially active against all the tested bacteria except <em>V. cholera.</em></p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"270\" src=\"/media/article_images/2024/06/14/178-1585380993-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1.</strong> Comparison of zone of inhibition of acetone extracts against tested microorganisms.</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"222\" src=\"/media/article_images/2024/06/14/178-1585380993-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2.</strong> Comparison of zone of inhibition of ethanol extracts against tested microorganisms.</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"figure3\">\r\n<figure class=\"image\"><img alt=\"\" height=\"279\" src=\"/media/article_images/2024/06/14/178-1585380993-Figure3.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 3. </strong>Potential zone of inhibitions obtained from acetone and ethanol extracts of selected samples: a. <em>S. cumini : S. typhi</em>, b.<em> L. chinensis</em>:<em> S. aureus, </em>c.<em> P. dactylifera: P. aeruginosa, </em>d. <em>A. heterophyllus: Micrococcus</em> spp., e.<em> S. cumini: S. aureus, </em>f.<em> L. Chinensis</em>: <em>E. coli,</em> g.<em> P. dactylifera: P. aeruginosa,</em> h. <em>A. heterophyllus: V. cholerae </em>(Figure a-d: acetone extracts, Figure e-h: ethanol extracts).</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"Table-2\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1585380993-table2/\">Table-2</a><strong>Table 2. </strong>Zone of inhibition of the samples against gram positive and gram-negative bacteria.</p>\r\n</div>\r\n\r\n<div id=\"Table-3\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1585380993-table3/\">Table-3</a><strong>Table 3. </strong>Minimum inhibitory concentration of tested samples against different microorganisms.</p>\r\n</div>\r\n\r\n<div id=\"Table-4\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1585380993-table4/\">Table-4</a><strong>Table 4.</strong> Minimum bactericidal concentration of tested samples against different microorganisms.</p>\r\n</div>\r\n\r\n<div id=\"Table-5\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1585380993-table5/\">Table-5</a><strong>Table 5. </strong>MIC Index (<sup>MBC</sup>/<sub>MIC</sub>) of tested samples against different microorganisms.</p>\r\n</div>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Effect on cytotoxic activity by brine shrimp lethality test</strong><br />\r\nExtracts exhibit LC<sub>50</sub> ≤ 30 µg/ml have significant cytotoxic activities, LC<sub>50</sub> ˃30 and LC<sub>50</sub> ≤100 possess mild toxicity and LC<sub>50</sub> ˃ 100 can be considered low toxic [<a href=\"#r-14\">14</a>]. Only seeds of P. dactylifera showed mild cytotoxicity where LC<sub>50</sub> values of acetone extracts and ethanol extracts were 69.42 and 68.19 µg/ml respectively. All the other samples were found relatively less toxic as LC<sub>50</sub> value more than 100 µg/ml (<a href=\"#figure4\">Figure 4</a>, <a href=\"#figure5\">5 </a>and <a href=\"#figure6\">6</a>). The LC<sub>50</sub> values were obtained (<a href=\"#figure6\">Figure 6</a>) by plotting log concentration vs percent shrimp mortality (<a href=\"#figure4\">Figure 4</a> and <a href=\"#figure5\">5</a>) into MS Excel and generating linear regression equation: Y = a + bX, Y – the dependent variable, a – the y-intercept, b – the slope of the line, and X – the independent/ explanatory variable. The dotted lines in the graph were plotted to produce regression equation for calculating LC<sub>50</sub> values (<a href=\"#figure4\">Figure 4</a> and <a href=\"#figure5\">5</a>). The values in Figure 6 were obtained from the regression equation as per plot of <a href=\"#figure4\\\">Figure 4</a> and<a href=\"#figure5\"> 5</a>.</p>\r\n\r\n<div id=\"figure4\">\r\n<figure class=\"image\"><img alt=\"\" height=\"300\" src=\"/media/article_images/2024/06/14/178-1585380993-Figure4.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 4.</strong> Log concentration vs percent shrimp mortality of acetone extracts of the tested samples.</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"figure5\">\r\n<figure class=\"image\"><img alt=\"\" height=\"284\" src=\"/media/article_images/2024/06/14/178-1585380993-Figure5.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 5. </strong>Log concentration vs percent shrimp mortality of ethanol extracts of the tested samples.</figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"figure6\">\r\n<figure class=\"image\"><img alt=\"\" height=\"310\" src=\"/media/article_images/2024/06/14/178-1585380993-Figure6.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 6. </strong>Comparison of LC<sub>50</sub> values of acetone and ethanol extracts (µg/ml) of the samples.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>Among all the samples, <em>S. cumini</em> was found more active compared to other samples. The best result obtained from acetone extract of <em>S. cumini</em> against <em>S. typhi</em> and <em>E. coli</em> (<a href=\"#figure1\">Figure 1</a> and <a href=\"#figure3\">3</a>). Our study, in accordance with another previous study where the MIC value of the seed extract against <em>E. coli, P. aeruginosa</em> was as 6.25 mg/ml whereas against <em>S. aureus</em> MIC value was 3.13 mg/ml [<a href=\"#r-15\">15</a>] suggests antibacterial potentialities of seeds of <em>S. cumini</em>. Seeds of <em>L. chinensis</em> are used in different countries of Asia for the treatment of bacterial infection and the related diseases [<a href=\"#r-16\">16</a>]. Our study supports them and provides scientific prove as both acetone and ethanol extract exhibited good antibacterial function. Our study, in consistent with another previous study of ours [<a href=\"#r-17\">17</a>] confirms antibacterial potentialities of seeds of <em>P. dactylifera</em>. Seeds of <em>A. heterophyllus </em>was found to be active against <em>V. cholerae </em>as the ethanolic extract exhibited bactericidal effects. Thus, it can be used to treat cholera diseases. A study mentioned that extract from the seeds are useful in the treatment of diarrhea [<a href=\"#r-18\">18-19</a>]. Our study also suggests that seeds of <em>A. heterophyllus</em> can be a source of remedy for severe watery diarrhea. The difference of the quality and quantity of plant ingredients, responsible for the bioactivity could be because of the diversified secondary metabolites (either constitutive or inducible: might function as antibacterial and cytotoxic agent) present within the extracts and the solvents used [<a href=\"#r-20\">20</a>]. Different types of polar solvents possess different ranges of soluble capacity for various phytoconstituents. Plant and/or plant parts act as the profound packs or reservoir of secondary metabolites [<a href=\"#r-21\">21-22</a>].<br />\r\nIn the study we found two trends of results: in some cases, extracts were found potential against gram positive bacteria and in other cases extracts were found more active against gram negative bacteria. Structure of the membrane of gram-positive bacteria includes low capacity of buffering against protonation generated by plant compounds. Thus, intra-cellular space of bacteria become highly acidified, lessening metabolic energy generation of bacteria. The outer polysaccharide layer consisting of lipid and extra components of minor membranes provide greater capability of buffering to gram negative bacteria, which function as protective wall against hydrophobic compounds. Therefore, gram negative bacteria are lesser sensitive to antibacterial properties. Here, gram negative bacteria showed more susceptibility than gram positive bacteria. The compound that can efficiently fuse the bilayer of lipid with increased fluid passing capability via cellular membrane is considered potential to show the activity as antibacterial agent [<a href=\"#r-23\">23, 24, 25</a>].<br />\r\nResistance to antibiotic as a huge concern for global public health [<a href=\"#r-26\">26</a>]. In a study conducted in one referral center in 2005 a whole of 57% of <em>S. typhi</em> strains isolated were MDR (multi drug resistant) and NAR (nalidaxic acid resistant) [<a href=\"#r-27\">27</a>]. Methicillin-resistant <em>S. aureus</em> could also be a significant cause of human infections. It is a most wanted explanation for bacteremia, infective endocarditis, osteoarticular, pleuropulmonary, and device-related infections [<a href=\"#r-28\">28</a>]. As antibiotic resistance is a significant problem worldwide, a replacement source of treating bacterial infections could also be a crying need. In line with our study, seeds of the four common summer fruits could be a potential source to isolate bioactive compounds [<a href=\"#r-29\">29-32</a>] that might heal the multi-drug resistant problems worldwide. As a whole, all the four samples can be a candidate for extracting antibacterial and cytotoxic bioactive agents from them.</p>"
},
{
"section_number": 5,
"section_title": "CONCLUSIONS",
"body": "<p>Seeds of many plants are utilized by traditional healers as drug and therapeutics to treat different diseases and infections. But most of the times seeds of plants are used only for sowing purposes, more importantly to be noted, sometimes seeds are thrown away as wastes, rather than utilizing or understanding their potentiality as bioactive compounds. More research is needed to evaluate the potentiality of seeds in this regard. In this <em>in vitro </em>study of seeds of four common summer fruits of Bangladesh, provide scientific footing to enhance confidence on the traditional claims of seeds. The antibacterial and cytotoxic activity of tested seed extracts recommends further isolation, purification and characterization of active constituents through bioassays, i<em>n-vivo </em>trials and other state-of art technology, leading to sort out the outcome of the effects of bioactive compounds and its role as therapeutics and pharmaceuticals in health care system, as well.</p>"
},
{
"section_number": 6,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>The authors express thanks to Khulna University Research Cell (KURC), Khulna University, Bangladesh for providing the grant to conduct this fundamental research. The authors also express thanks to their respective institute: Khulna University, Bangladesh.</p>"
},
{
"section_number": 7,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>MSA was involved in conception and design of the experiments. SAR contributed to perform the experiments. SAR and MSA analyzed data and contributed to drafting the article. SAR and MSA contributed to revising it critically for important intellectual content. MSA made the final approval of the version to be published.</p>"
},
{
"section_number": 8,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/06/14/178-1585380993-Figure1.jpg",
"caption": "Figure 1. Comparison of zone of inhibition of acetone extracts against tested microorganisms.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/06/14/178-1585380993-Figure2.jpg",
"caption": "Figure 2. Comparison of zone of inhibition of ethanol extracts against tested microorganisms.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/06/14/178-1585380993-Figure3.jpg",
"caption": "Figure 3. Potential zone of inhibitions obtained from acetone and ethanol extracts of selected samples: a. S. cumini : S. typhi, b. L. chinensis: S. aureus, c. P. dactylifera: P. aeruginosa, d. A. heterophyllus: Micrococcus spp., e. S. cumini: S. aureus, f. L. Chinensis: E. coli, g. P. dactylifera: P. aeruginosa, h. A. heterophyllus: V. cholerae (Figure a-d: acetone extracts, Figure e-h: ethanol extracts).",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/06/14/178-1585380993-Figure4.jpg",
"caption": "Figure 4. Log concentration vs percent shrimp mortality of acetone extracts of the tested samples .",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/06/14/178-1585380993-Figure5.jpg",
"caption": "Figure 5. Log concentration vs percent shrimp mortality of ethanol extracts of the tested samples.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/06/14/178-1585380993-Figure6.jpg",
"caption": "Figure 6. Comparison of LC50 values of acetone and ethanol extracts (µg/ml) of the samples.",
"featured": false
}
],
"authors": [
{
"id": 856,
"affiliation": [
{
"affiliation": "Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh."
}
],
"first_name": "Sheikh Ashikur",
"family_name": "Rahman",
"email": null,
"author_order": 1,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 194
},
{
"id": 857,
"affiliation": [
{
"affiliation": "Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh."
}
],
"first_name": "Md. Shamim",
"family_name": "Akhter",
"email": "shamim11akhter@bge.ku.ac.bd",
"author_order": 2,
"ORCID": null,
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Md. Shamim Akhter, Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh. E-mail: shamim11akhter@bge.ku.ac.bd",
"article": 194
}
],
"views": 684,
"downloads": 125,
"references": [
{
"id": 6556,
"serial_number": 1,
"pmc": null,
"reference": "Li F, Wang Y, Li D, Chen Y, Dou QP. Are we seeing a resurgence in the use of natural products for new drug discovery? Expert Opin Drug Discov. 2019; 14: 417–20. doi:10.1080/17460441.2019.1582639.",
"DOI": null,
"article": 194
},
{
"id": 6557,
"serial_number": 2,
"pmc": null,
"reference": "Blockley A, Elliott D, Roberts A, Sweet M. Symbiotic Microbes from Marine Invertebrates: Driving a New Era of Natural Product Drug Discovery. Diversity 2017; 9: 49. doi:10.3390/d9040049.",
"DOI": null,
"article": 194
},
{
"id": 6558,
"serial_number": 3,
"pmc": null,
"reference": "Al-Snafi AE. Therapeutic importance of Hyoscyamus species grown in Iraq (Hyoscyamus albus, Hyoscyamus niger and Hyoscyamus reticulates)-A review. IOSR J. Pharm. 2018; 8:6:18-32.",
"DOI": null,
"article": 194
},
{
"id": 6559,
"serial_number": 4,
"pmc": null,
"reference": "Shah Z, Shafi S. Diabetes the Global Economic Burden of Adults and the Role of Herbalism as a safe & altenative cost-effective therapy. An Updated Overview. J. drug deliv. ther. 2019; 9:3: 954-961. doi:10.22270/jddt.v9i3-s.3085.",
"DOI": null,
"article": 194
},
{
"id": 6560,
"serial_number": 5,
"pmc": null,
"reference": "Sutradhar KB, Saha A, Huda NH, Uddin, R. Irrational use of antibiotics and antibiotic resistance in southern rural Bangladesh: perspectives from both the physicians and patients. Annu Res Rev Biol. 2014; 1421-1430. doi:10.9734/ARRB/2014/8184.",
"DOI": null,
"article": 194
},
{
"id": 6561,
"serial_number": 6,
"pmc": null,
"reference": "Faiz MA, Basher A. Antimicrobial resistance: Bangladesh experience. In Regional Health Forum 2011; 15:1:1-8.",
"DOI": null,
"article": 194
},
{
"id": 6562,
"serial_number": 7,
"pmc": null,
"reference": "Datir SS. Plant Metabolites as New Leads to Anticancer Drug Discovery: Approaches and Challenges. In Anticancer Plants: Natural Products and Biotechnological Implements 2018; 141-161. doi:10.1007/978-981-10-8064-7_7.",
"DOI": null,
"article": 194
},
{
"id": 6563,
"serial_number": 8,
"pmc": null,
"reference": "Pavithra PS, Janani VS, Charumathi KH, Indumathy R, Potala S, Verma RS. Antibacterial activity of plants used in Indian herbal medicine. Int. J. Green Pharm. 2010; 4:1. doi:10.22377/ijgp.v4i1.114.",
"DOI": null,
"article": 194
},
{
"id": 6564,
"serial_number": 9,
"pmc": null,
"reference": "Rahman MS, Begum B, Chowdhury R, Rahman KM, Rashid MA. Preliminary cytotoxicity screening of some medicinal plants of Bangladesh. Dhaka Univ. J. Pharm. Sci. 2008; 7:1: 47-52. doi:10.3329/dujps.v7i1.1217.",
"DOI": null,
"article": 194
},
{
"id": 6565,
"serial_number": 10,
"pmc": null,
"reference": "Mukul SA, Biswas SR, Rashid AM. Biodiversity in Bangladesh. In Global Biodiversity, Volume 1: Selected Countries in Asia 2018; 93-107.",
"DOI": null,
"article": 194
},
{
"id": 6566,
"serial_number": 11,
"pmc": null,
"reference": "Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. Pharm. Anal. 2016; 6:2:71-79. doi:10.1016/j.jpha.2015.11.005.",
"DOI": null,
"article": 194
},
{
"id": 6567,
"serial_number": 12,
"pmc": null,
"reference": "Mariselvam R, Ranjitsingh AJA, Selvakumar PM, Krishnamoorthy, R, Alshatwi Eco friendly natural dyes from Syzygium cumini (L) (Jambolan) fruit seed endosperm and to preparation of antimicrobial fabric and their washing properties. Fibers Polym. 2017; 18:3: 460-464. doi:10.1007/s12221-017-1097-6.",
"DOI": null,
"article": 194
},
{
"id": 6568,
"serial_number": 13,
"pmc": null,
"reference": "Tippett LO, Zeleznick LD, Robb CA. Modification of the microtiter technique for antimicrobial drug susceptibility testing by incorporation of indicators. Appl. Environ. Microbiol. 1970; 20:3:342-345.",
"DOI": null,
"article": 194
},
{
"id": 6569,
"serial_number": 14,
"pmc": null,
"reference": "Meyer BN, Ferrign, RN, Putnam, JE, Jacobson, LB, Nicholas, DE, McLaughlin, JL. Brine shrimp: A convenient general bioassay for active plant constituents. Planta Med. 1982; 45:31-34.",
"DOI": null,
"article": 194
},
{
"id": 6570,
"serial_number": 15,
"pmc": null,
"reference": "Banerjee J, Narendhirakannan RT. Phytochemical analyses, antibacterial, in vitro antioxidant and cytotoxic activities of ethanolic extract of Syzygium cumini (L.) seed extract. Int. J. Pharm. Sci. 2011; 2:7: 1799.",
"DOI": null,
"article": 194
},
{
"id": 6571,
"serial_number": 16,
"pmc": null,
"reference": "Ibrahim SR, Mohamed GA. Litchi chinensis: medicinal uses, phytochemistry, and pharmacology. J Ethnopharmacol. 2015; 174:492-513. doi:10.1016/j.jep.2015.08.054.",
"DOI": null,
"article": 194
},
{
"id": 6572,
"serial_number": 17,
"pmc": null,
"reference": "Aamir J, Kumari A, Khan MN, Medam, SK. Evaluation of the combinational antimicrobial effect of Annona Squamosa and Phoenix Dactylifera seeds methanolic extract on standard microbial strains. Int Res. J Biol Sci. 2013; 2:5: 68-73.",
"DOI": null,
"article": 194
},
{
"id": 6573,
"serial_number": 18,
"pmc": null,
"reference": "Hari A, Revikumar KG, Divya D. Artocarpus: A review of its phytochemistry and pharmacology. J. Pharm. Res. 2014; 9:1:7-12.",
"DOI": null,
"article": 194
},
{
"id": 6574,
"serial_number": 19,
"pmc": null,
"reference": "Debnath S, Rahman SH, Deshmukh G, Duganath N, Pranitha C, Chiranjeevi A. Antimicrobial screening of various fruit seed extracts. Phcog J. 2011; 3:19:83-86. doi:10.5530/pj.2011.19.15.",
"DOI": null,
"article": 194
},
{
"id": 6575,
"serial_number": 20,
"pmc": null,
"reference": "Umaarasu T, Padmavathy K, Thirunavukkarasu D, Rajesh SV, Govindaraj J, Shanmugam G. Evaluation of the antimicrobial efficacy and phytochemical constituents of the extracts of Andrographis paniculata against drug-resistant bacterial pathogens. Drug Invent. Today. 2019: 11:10.",
"DOI": null,
"article": 194
},
{
"id": 6576,
"serial_number": 21,
"pmc": null,
"reference": "Gopalakrishnan S, Rajameena R, Vadivel E. Antimicrobial activity of the leaves of Myxopyrum serratulum AW Hill. Int. J. Pharm. Sci. Drug. Res 2012; 4:31-34.",
"DOI": null,
"article": 194
},
{
"id": 6577,
"serial_number": 22,
"pmc": null,
"reference": "Rahman SA, Akhter, MS. Antibacterial and cytotoxic activity of seeds of white hyacinth bean (Lablab purpureus sweet ‘white’). J Adv Biotechnol Exp Ther. 2018; 1:2: 49-54.",
"DOI": null,
"article": 194
},
{
"id": 6578,
"serial_number": 23,
"pmc": null,
"reference": "Rakholiya K, Kaneria M, Desai D, Chanda S. Antimicrobial activity of decoction extracts of residual parts (seed and peels) of Mangifera indica L. var. Kesar against pathogenic and food spoilage microorganism. Microbial pathogens and strategies for combating them: Science, Technology and Education. FORMATEX 2013; 850-6.",
"DOI": null,
"article": 194
},
{
"id": 6579,
"serial_number": 24,
"pmc": null,
"reference": "Asif A, Farooq U, Akram K, Hayat Z, Shafi A, Sarfraz F, Aftab S. Therapeutic potentials of bioactive compounds from mango fruit wastes. Trends Food Sci Tech. 2016; 53:102-112. doi:10.1016/j.tifs.2016.05.004.",
"DOI": null,
"article": 194
},
{
"id": 6580,
"serial_number": 25,
"pmc": null,
"reference": "Ediriweera MK, Tennekoon KH, Samarakoon SR. A review on ethnopharmacological applications, pharmacological activities, and bioactive compounds of Mangifera indica (mango). Evid. Based Complementary Altern. Med. 2017. doi:10.1155/2017/6949835",
"DOI": null,
"article": 194
},
{
"id": 6581,
"serial_number": 26,
"pmc": null,
"reference": "Aslam B, Wang W, Arshad MI, Khurshid M, Muzammil S, Rasool MH, Salamat MKF. Antibiotic resistance: a rundown of a global crisis. Infect Drug Resist. 2018; 11:1645. doi: 10.2147/IDR.S173867.",
"DOI": null,
"article": 194
},
{
"id": 6582,
"serial_number": 27,
"pmc": null,
"reference": "Hoque R, Mostafa A, Haque M. Intern doctors’ views on the current and future antibiotic resistance situation of Chattagram Maa O Shishu Hospital Medical College, Bangladesh. Ther Clin Risk Manag. 2015; 11:1177. doi: 10.2147/TCRM.S90110",
"DOI": null,
"article": 194
},
{
"id": 6583,
"serial_number": 28,
"pmc": null,
"reference": "Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev. 2015; 28:3:603-661. doi: 10.1128/CMR.00134-14.",
"DOI": null,
"article": 194
},
{
"id": 6584,
"serial_number": 29,
"pmc": null,
"reference": "Martindale WH. Essential oil in relation to their antiseptic powers. Purfum. Essent. Oil Rec. 1910; 1: 266274.",
"DOI": null,
"article": 194
},
{
"id": 6585,
"serial_number": 30,
"pmc": null,
"reference": "Hoffmann C, Evans AC. The Use of Spices as Preservatives. Indus & Engin Chem. 1911; 3(11): 835-838.",
"DOI": null,
"article": 194
},
{
"id": 6586,
"serial_number": 31,
"pmc": null,
"reference": "Newman DJ, Cragg GM. Natural products as sources of new drugs over the 30 years from 1981 to 2010. J of nat. prod. 2012; 75(3): 311-335.",
"DOI": null,
"article": 194
},
{
"id": 6587,
"serial_number": 32,
"pmc": null,
"reference": "Shen B. A new golden age of natural products drug discovery. Cell. 2015; 163(6): 1297-1300.",
"DOI": null,
"article": 194
}
]
},
{
"id": 190,
"slug": "178-1579228061-pharmacological-effect-of-methanolic-and-hydro-alcoholic-extract-of-coconut-endocarp",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1579228061",
"recieved": "2020-01-17",
"revised": null,
"accepted": "2020-04-06",
"published": "2020-06-01",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/35/178-1579228061.pdf",
"title": "Pharmacological effect of methanolic and hydro-alcoholic extract of Coconut endocarp",
"abstract": "<p><em>Cocos nucifera </em>used as natural remedies in a wide variety of diseases. The current experiment aimed to determine the qualitative phytochemicals, anxiolytic, anti-diarrheal, anti-inflammatory, thrombolytic and cytotoxic actions of <em>C. nucifera</em> endocarp, which extracted by using methanol (MeOH-CNE) and the hydro-alcohol (HaE-CNE). The MeOH-CNE and HaE-CNE subjected to phytochemical screening, where both extracts showed the existence of secondary metabolites such as carbohydrates, flavonoids, cardiac glycosides, and proteins. The anxiolytic activity screened by elevated plus maze experiment, whereas the percentage of open arm accounts 74.55 ± 4.54%, 66.31 ± 4.41% at 400 mg/kg for HaE-CNE, MeOH-CNE respectively (<em>P < </em>0.05) while the standard drug Diazepam (75.24 ± 3.91%). In the anti-diarrheal test, extracts (200 and 400 mg/kg) and standard drug loperamide (5 mg/kg) showed dose-dependent significant (<em>P <</em> 0.05) inhibition against castor oil-induced diarrhea. The MeOH-CNE and HaE-CNE exhibited 72.91 ± 4.20 %, 64.42 ± 5.50% inhibition of protein denaturation at 500 μg/ml, while in the thrombolytic test, HaE-CNE showed the highest and significant (<em>P <</em> 0.05) in clot lysis activity on human blood in comparison to water, whereas streptokinase used as standard. In brine shrimp lethality bioassay, the LC<sub>50 </sub>of HaE-CNE and MeOH-CNE ware 432.35 µg/ml and 1173.88 µg/ml respectively. The LC<sub>50</sub> for standard vincristine sulfate was 43.15 µg/ml. The current results suggested that MeOH-CNE and HaE-CNE have promising pharmacological activity.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 171-181.",
"academic_editor": "Dr. Md. Abdul Hannan, Dongguk University, South Korea.",
"cite_info": "Akter S, Shah M, et al. Pharmacological effect of methanolic and hydro-alcoholic extract of Coconut endocarp. J Adv Biotechnol Exp Ther. 2020; 3(3): 171-181.",
"keywords": [
"Anxiolytic",
"Antidiarrheal",
"Cocos nucifera",
"Endocarp",
"Cytotoxic activity"
],
"DOI": "10.5455/jabet.2020.d123",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p><em>Cocos nucifera</em> (L.) belongs to the family Arecaceae (palm family), which is frequently termed a tree of life. For many years, coconut products have used as folk medicine. In ayurvedic medicine, coconut products such as oil, milk, cream, and water are being castoff in the treatment of hair loss, burns and heart complications [<a href=\"#r-1\">1</a>]. All parts of <em>C. nucifera</em> can be used traditionally in various pathological conditions such as shell fibers of <em>Cocos nucifera</em> are used in diarrhea [<a href=\"#r-2\">2</a>],<em> </em>antipyretic, renal inflammation and as a cream for dermatitis, sores, and injuries. The fibers are also used in asthma and diabetes. Moreover, the leaves and roots parts are used in diarrhea and stomach pains. The solid albumen of coca extracted as oil, pulp, milk which are used in antipyretic, diarrheal treatment, preventing hair loss, wound healing [<a href=\"#r-3\">3, 4</a>]; oral contraceptive, diarrheal treatment, aphrodisiac, get rid of from skin rash affected by HIV infection, respectively. The white surface of the Coca is used to consider for fever and malaria. The water of <em>C. nucifera</em> is used in treatment of renal disease [<a href=\"#r-4\">4-7</a>]. Several pharmacological and biological activities are identified in several reports, including analgesia; anti-inflammatory, antimicrobial, antioxidant, anti-osteoporosis, anti-diabetic, anti-neoplastic, anthelminthic, antihypertensive, vasodilation, defense of kidney, heart, and liver functions and anti-malarial activities [<a href=\"#r-8\">8-12</a>].<br />\r\nAnxiety syndromes are the most usual mental, emotional, and behavioral complications [<a href=\"#r-13\">13</a>] affecting 264 million peoples around the globe [<a href=\"#r-14\">14</a>]. In terms of overall management and treatment, depression among mood disorders has always been problematic [<a href=\"#r-15\">15</a>]. Stress, depression and anxiety might trigger insomnia. Medicinal plants have been taken as sleep aids in insomnia throughout the world [<a href=\"#r-16\">16</a>]. Despite the development of more molecules against the treatment of depression, unfortunately this disease remains untreated in many patients because of the burden of their side effects [<a href=\"#r-17\">17</a>]. Therefore, searching for new therapeutic agents having no adverse effects may be the option of improved pharmacological actions, higher efficacy and safety.<br />\r\nDiarrheal syndromes are a key problem in underdeveloped countries and are accountable for the loss of a huge number of people every year [<a href=\"#r-18\">18</a>]. Diarrhea is characterized by an increase in the water content, volume, or incidence of stools [<a href=\"#r-19\">19</a>]. The research shown that several microbes like <em>Salmonella</em>, <em>Escherichia coli, Vibrio cholerae</em> and <em>Shigella</em> generate and release enterotoxins which are the major cause of diarrhea in developing countries [<a href=\"#r-20\">20</a>]. Opioids and its derivatives such as difenoxin and loperamide are mostly used in the management of diarrhea [<a href=\"#r-21\">21</a>]. Medicinal plants are an effective source for developing new antidiarrheal drugs. Therefore, the WHO has encouraged using medicinal plants in the management of diarrhea [<a href=\"#r-22\">22</a>]. Currently available opioids and its derivatives are linked with several adverse effects for example abdominal pain, distention, bloating, nausea, vomiting, and constipation [<a href=\"#r-23\">23</a>] and also cardiotoxicity has been testified by loperamide [<a href=\"#r-24\">24</a>].<br />\r\nEndocarp of <em>C. nucifera</em> is the hardest part of the fruit and a rich source of phenolic and flavonoid content [<a href=\"#r-25\">25</a>]. According to the literature review, <em>C. nucifera </em>shell has phenolic content [<a href=\"#r-26\">26</a>]. The study aimed to investigate the <em>in vivo</em> anxiolytic, antidiarrheal and <em>in vitro</em> anti-inflammatory and thrombolytic, cytotoxic effects along with the phytochemical study of methanolic and hydro-alcoholic extract of <em>C. nucifera</em> endocarp. The present study investigates the <em>in vivo</em> anxiolytic, antidiarrheal and <em>in vitro</em> anti-inflammatory, thrombolytic, cytotoxic effects of <em>C. nucifera</em>, a Bangladeshi plant for the first time.</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Chemicals</strong><br />\r\nLoperamide hydrochloride, castor oil (Sigma-Aldrich, MO, USA), Streptokinase (Beacon Pharmaceutical Ltd., Mymensingh, Bangladesh), Sea salt non ionized NaCl, vincristine sulphate (Sigma-Aldrich, MO, USA), ethanol, Tween 80 were used in this study. All other chemicals and reagent were used of analytical grade.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Preparation of plant extract</strong><br />\r\nThe moist endocarp of <em>C. nucifera</em> were collected from the Sitakunda local area of Chittagong, Bangladesh, which later validated by Professor Dr. Sheikh Bokhtear Uddin, Department of Botany, University of Chittagong. The <em>C. nucifera</em> endocarp layer was collected in a fresh condition. Then these are cut into small pieces if necessary to make it suitable for grinding purposes. Then the crude parts were rubbed for ten days and later grounded to powder by a mechanical drier (Ecocell, MMM Group, Germany) at 55-60 °C. By using another mechanical grinder (NOWAKE, Japan), the samples were ground to a coarse powder. The powder (500 g.) was soaked in 500 ml of methanol and 70% ethanol, respectively, for a week with occasional shaking and stirring on a shaker machine at room temperature. It was then filtered through a cotton plug followed by Whatman filter paper No. 1. After filtration, the filtrate was evaporated by using a rotary evaporator at 50 °C under reduced pressure to obtain the methanol crude extract (MeOH-CNE) (9.5 g) and hydroalcoholic extract (HaE-CNE) (11.48 g). The extracts were conserved at 4 °C in a refrigerator until further use.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Experimental animals</strong><br />\r\nSwiss Albino mice (Six-seven weeks old) of either sex were purchased from the International Center for Diarrheal Diseases Research, Dhaka, Bangladesh (ICDDRB). The animals were adapted to the laboratory condition (25 ± 2 °C with a light/dark cycle of 12 h) for seven days before the study. The study was conducted following approval by the Institutional Animal Ethical Committee, Department of Pharmacy, International Islamic University Chittagong, Bangladesh, under the reference number (P&D-147/13-18).</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Phytochemical screening</strong><br />\r\nThe qualitative phytochemical screening was executed according to the regular procedures [<a href=\"#r-27\">27</a>] and the results shown presence or absence of secondary plant metabolites such as alkaloids, flavonoids, tannins, saponins, phenol, carbohydrate and glycoside.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Anxiolytic activity</strong><br />\r\n<em>Elevated plus maze test</em><br />\r\nAnxiolytic activity assessed by an elevated plus maze (EPM) apparatus, which used for the unlearned response. The EPM apparatus elevated from the floor at 40 cm, where it consists of two open arms and closed arms with a central square [<a href=\"#r-28\">28, 29</a>]. The negative control received 1% Tween-80 (10 ml/kg, b.w), the reference drug Diazepam received (1 mg/kg, b.w) intraperitoneally, while the treatment group received 200 mg/kg and 400 mg/kg, b.w. by oral gavage. After sixty minutes, each mouse positioned in the center of the EPM facing towards the closed arm. Entry into open and closed arm recorded for 5 minutes and calculated the percentage by the resulting equation: % of open arm entries=(Number of entries in open arm)/(Number of entries in open arm+number of entries in closed arm ) ×100</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Anti-diarrheal activity</strong><br />\r\n<em>Castor oil-induced diarrhea</em><br />\r\nThe castor-oil induced diarrhea was followed to evaluate the antidiarrheal according to the method described by Taufiq et al. and Bellah et al. [<a href=\"#r-30\">30, 31</a>]. Mice of either sex fasted for 24 hours. The negative control received 1% Tween-80 (10 ml/kg, b.w), the reference drug loperamide (5 mg/kg, b.w; as oral suspension), while the treatment group received 200 mg/kg and 400 mg/kg, b.w. by oral gavage. After 60 minutes, each mouse received 0.5 ml castor oil orally by gavage and individually placed in cages consist of transparent paper. The transparent paper changed in every hour and counted dry and wet feces in every 60 min for 4 hours. The equation calculated the level of % inhibition of defecation: % inhibition of defecation= (A-B)/A ×100<br />\r\nWhere, A = average eradication feces number of the control group; B = average eradication feces number of the text group.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Anti-inflammatory activity</strong><br />\r\n<em>Inhibition of protein denaturation method</em><br />\r\nThe anti-inflammatory activity of the extracts determined using the protein denaturation method [<a href=\"#r-32\">32, 33</a>]. The constituents of the reaction solution were 100 μl of the extracts (final concentration 62.5-500 μg/ml) and 100 μl of 5% aqueous bovine serum albumin. Then the pH was adjusted using glacial acetic acid. The samples were incubated at 37 °C for 20 min and then heated to 70 °C for 10 min. After incubation, the mixture was allowed to cool for 10 min, and a turbid solution found. Then the turbidity was measured at 660 nm. The blank consist of the sample and distilled water. Distilled water was used as negative control. The positive control was diclofenac sodium. Percentage inhibition was calculated using the formula:<br />\r\n% of protein denaturation = [(absorbance of control – absorbance of test sample) / (absorbance of test control)] × 100</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>Thrombolytic activity</em><br />\r\nThe clot lysis activity performed as described by Prasad et al. [<a href=\"#r-34\">34</a>]. A 3 ml venous blood is withdrawn from the ten healthy volunteers who did not have any previous history of taking NSAID or contraceptives. The withdrawn blood was distributed in nine different pre-weighed sterile micro centrifuge tubes (0.5 ml/tube) and incubated for 45 min at 37 °C. After the formation of the clot, completely removed the serum without disturbing the clot and each tube reweighed for calculating the clot weight. The plant extracts ware added to each micro centrifuge tube separately containing pre-weighed clot. 100 µL of streptokinase and distilled water were added separately to the positive and negative control group. All the tubes were then incubated at 37 °C for 90 min and observed for clot lysis. The released fluid was removed and reweighed the tube to calculate the difference in weight after clot disruption.<br />\r\n% of clot lysis = (weight of clot after remove of fluid/clot weight) × 100</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>Brine shrimp cytotoxicity</em><br />\r\nThe brine shrimp lethality bioassay experiment was followed by the previous reported method [<a href=\"#r-35\">35</a>]. In the artificial seawater (3.8% NaCl solution), the shrimp eggs hatched for 48 hours for maturing the shrimp called nauplii. The shrimp egg collected from the Katabon, Dhaka. The crude extract was dissolved in DMSO to obtain a solution of 5 mg/ml, which was subjected to serially diluted concentrations of 20 to 100 μg/ml, followed by the addition of 5.0 ml of artificial seawater in each test tubes. Ten of the living nauplii applied to each of all experimental vials and control vials. Following 24 hours, all vials inspected by an amplifying glass, and the number of living nauplii in each vial was observed and recorded. Experiments were conducted in a set of triplicate manner along with reference drug vincristine sulfate. The lethal concentration (LC<sub>50</sub>) that would kill one-half of the nauplii was determined from a linear regression equation.<br />\r\n% of mortality = (N<sub>0</sub>-N<sub>1</sub>/N<sub>0</sub>) ×100<br />\r\nWhere, N<sub>0</sub>= the number of nauplii taken; N<sub>1</sub>= the number of nauplii alive.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Statistical analysis</strong><br />\r\nResult represented as Mean ± Standard Error Mean (SEM). P (< 0.05 and < 0.001) were measured as statistically significant while the Dunnet’s test was used to describe the significance using GraphPad Prism Version 6.0 (GraphPad software Inc., San Diego, CA).</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><em><strong>Phytochemical </strong></em><strong><em>analysis revealed the presence of secondary metabolites</em></strong><br />\r\nThe qualitative phytochemical screenings of MeOH-CNE and HaE-CNE have performed to find out the presence or absence of secondary plant metabolites. In our results, both extracts showed the presence of carbohydrates, flavonoids, cardiac glycosides, and protein. Whereas, only HaE-CNE showed the presence of alkaloids (<a href=\"#Table-1\">Table 1</a>).</p>\r\n\r\n<div id=\"Table-1\">\r\n<p><a href=\"https://jabet.bsmiab.org/table/178-1579228061-table1/\">Table-1</a><strong>Table 1.</strong> Qualitative phytochemical screening of MeOH-CNE and HaE-CNE.</p>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Both the extracts reveled significant anxiolytic activity in EPM test</strong><br />\r\nIn elevated plus-maze (EPM) test, both extract reveled significant (<em>P <</em> 0.05) anxiolytic activity compared to the control group. The HaE-CNE treated mice at dose of 400 mg/kg showed significant (<em>P <</em> 0.05) increase of the percentage of open-arm entries (74.55 ± 4.54) compared to the control group, whereas the MeOH-CNE treated mice showed 66.31 ± 4.41 percentage of open-arm entries at the same dose (<a href=\"#figure1\">Figure 1</a>). On the other hand, the reference drug diazepam at a very small dose 1 mg/kg, b.w treated mice produced a noticeable increase in the percentages of open-arm entries (75.24 ± 3.91).</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"400\" src=\"/media/article_images/2024/26/17/178-1579228061-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1. </strong>Effect of MeOH-CNE and HaE-CNE on percentage of entry into open arm in elevated plus maze test in mice. Values are mean ± S.E.M. *<em>P <</em> 0.05 and **<em>P <</em> 0.01, significantly different from control; ANOVA followed Dunnett’s test (n = 6, per group). Where, MeOH-CNE: methanolic extract <em>Cocos nucifera </em>endocarp and HaE-CNE: hydro-alcoholic extract of <em>Cocos nucifera </em>endocarp.</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Both the extracts reduces the</strong> <strong>rate of defecation in a time-dependent manner</strong><br />\r\nIn the castor oil-induced diarrheal experiment, both extracts were found to be effective and significant (<em>P <</em> 0.05) in a dose dependent manner on experimental mice at all tested doses. At the dose of 400 mg/kg the HaE-CNE showed 56.79 ± 2.33% reductions in the rate of defecation in albino mice. Whereas, the MeOH-CNE extract showed 51.23 ± 3.26% reductions at the same dose. This condition was markedly reduced (62.49 ± 1.11%) by standard drug Loperamide at a dose of 5 mg/kg <a href=\"#figure2\">Figure 2</a>.</p>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"394\" src=\"/media/article_images/2024/26/17/178-1579228061-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2. </strong>Antidiarrheal activity of MeOH-CNE and HaE-CNE against castor oil-induced diarrhea in mice. All values are expressed as mean ± SEM (n = 6); Data were analyzed by one way analysis of variance using GraphPad Prism for Windows, Version 6.0 (GraphPad software Inc., San Diego, CA, USA) followed by Dunnett’s test for multiple comparisons,. Values with <sup>* </sup><em>P <</em> 0.05 were considered as significant. MeOH-CNE: methanolic extract <em>Cocos nucifera </em>endocarp and HaE-CNE: hydro-alcoholic extract of <em>Cocos nucifera </em>endocarp.</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Both the extracts increases the</strong> <strong>inhibition of protein denaturation</strong><br />\r\nThe results of anti-inflammatory activity of the extract are displayed in <a href=\"#figure3\">Figure 3</a>. The result showed a dose dependent and significantly (<em>P <</em> 0.05) increased the inhibition of protein denaturation by MeOH-CNE, HaE-CNE and Diclofenac-Na throughout the concentration range (62.5-500 μg/ml). The crude hydro alcoholic extract (HaE-CNE) demonstrated maximum inhibition of protein denaturation (72.91 ± 4.20%) at 500 μg/ml, whereas the methanolic extract (MeOH-CNE) showed 64.42 ± 5.50% inhibitions at the same concentration.</p>\r\n\r\n<div id=\"figure3\">\r\n<figure class=\"image\"><img alt=\"\" height=\"394\" src=\"/media/article_images/2024/26/17/178-1579228061-Figure3.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 3.</strong> Percentage inhibition of protein denaturation by MeOH-CNE, HaE-CNE. Results are mean ± SEM (n = 3). ANOVA followed by Dunnett’s test. Where, MeOH-CNE: methanolic extract <em>Cocos nucifera </em>endocarp and HaE-CNE: hydro-alcoholic extract of <em>Cocos nucifera </em>endocarp.</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Both the extracts</strong><strong> significantly increases clot lysis activity</strong><br />\r\nThe results of clot lysis activity of extracts are shown in <a href=\"#figure4\">Figure 4</a>. After addition of 100 μl of Streptokinase a positive control (15,00,000 I.U.) to the clots along with 90 minutes of incubation at 37 °C, showed 73.44 ± 1.87% clot lysis. When clots were treated with negative control (sterile distilled water), a negligible clot lysis (8.87 ± 1.32) has observed. After the treatment of clots with HaE-CNE and MeOH-CNE, the significant (<em>P <</em> 0.05) clot lysis was observed. The clot lysis activity of MeOH-CNE and HaE-CNE ware 35.73 ± 3.21% and 55.74 ± 2.78% respectively.</p>\r\n\r\n<div id=\"figure4\">\r\n<figure class=\"image\"><img alt=\"\" height=\"370\" src=\"/media/article_images/2024/26/17/178-1579228061-Figure4.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 4. </strong>Clot lysis by streptokinase, MeOH-CNE, HaE-CNE and water <em>in vitro</em>. Values are expressed as mean ± S.E.M. <em>P <</em> 0.05, significantly different from control; ANOVA followed by Dunnett’s test. Where, MeOH-CNE: methanolic extract <em>Cocos nucifera </em>endocarp and HaE-CNE: hydro-alcoholic extract of <em>Cocos nucifera </em>endocarp.</figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Both</strong><strong> the extracts significantly decreases the mortality of brine shrimp</strong><br />\r\nIn brine shrimp lethality bioassay, the rate of mortality of nauplii was presented in <a href=\"#figure5\">Figure 5</a>. The degree of lethality shown by the extracts was found to be directly proportional to the concentration of the extract ranging from the lowest concentration (20 µg/ml) to the highest concentration (1000 µg/ml). Both extract virtually non-toxic on the brine shrimp. They showed very low toxicity, giving LC<sub>50</sub> values greater than 100 μg/ml. The LD<sub>50</sub> values of MeOH-CNE, HaE-CNE and vincristine sulfate (VCS) were 432.35 µg/ml 1173.88 µg/ml and 43.15 µg/ml, respectively.</p>\r\n\r\n<div id=\"figure5\">\r\n<figure class=\"image\"><img alt=\"\" height=\"340\" src=\"/media/article_images/2024/26/17/178-1579228061-Figure5.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 5. </strong>Cytotoxic effects of MeOH-CNE and HaE-CNE. Extrapolation of inhibition concentrations of both MeOH-CNE, HaE-CNE and reference cytotoxic agent vincristine sulfate through regression analysis. Data are shown as mean ± SEM of 10 shrimps for each concentration. MeOH-CNE: methanolic extract <em>Cocos nucifera </em>endocarp and HaE-CNE: hydro-alcoholic extract of <em>Cocos nucifera </em>endocarp.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>The phytochemical analysis led on the plant extracts shown the presence of compound which is referred to show therapeutic as well as biological activities. Analysis of plant extract shows the presence of alkaloids, carbohydrates, flavonoids, cardiac glycosides, and protein. The phytochemical investigation led on the plant extracts shown the presence of compound which is referred to show therapeutic as well as physiological activities [<a href=\"#r-36\">36</a>]. The current findings are similar to the previous study of the ethanolic extract of mesocarp [<a href=\"#r-4\">4</a>].<br />\r\nThe elevated plus maze (EPM) test is considered to be a good test procedure to evaluate the anxiety-like behavior [<a href=\"#r-28\">28</a>, <a href=\"#r-37\">37</a>]. The increasing number of entries in the open arm indicates the anxiolytic effects and the increasing number of entries at the closed arm indicates the anxiogenic effects [<a href=\"#r-37\">37</a>, <a href=\"#r-38\">38</a>]. Administration of MeOH-CNE, HaE-CNE showed an increased amount of entries in the open arms, an indication of anxiolytic-like behavior. Similar observations found to the reference drug (diazepam) which significantly increased the percentage of open arm entries. The current findings are similar to the previous study of ethanolic extract of <em>C. nucifera</em> endocarp, whereas the 500 mg/kg dose exhibited 74.77 ± 6.86 percentages of open arm entries [<a href=\"#r-39\">39</a>].<br />\r\nThe assessment of <em>C. nucifera</em> endocarp on castor oil-induced diarrheal mice to exhibit the dose-dependent manner, especially HaE-CNE showed a significant (<em>P <</em> 0.05) amount of reduction of diarrhea in comparison to the negative control. Similar observations found to the reference drug (Loperamide) which significantly increased the reduction of diarrheal faces. Castor oil is a recognized diarrheal agent due to the presence of ricinoleic acid which causes changes in the intestinal mucosa, as a result, fluid and watery luminal content that flow rapidly by intestine [<a href=\"#r-40\">40</a>]. Due to the release of ricinoleic acid, it produces different inflammatory mediators such as prostaglandins, nitric oxide, platelet-activating factor, cAMP, and histamine [<a href=\"#r-41\">41, 42</a>]. The present result validating the traditional use and the presence of secondary metabolites flavonoids [<a href=\"#r-43\">43</a>].<br />\r\nThe assay of anti-inflammatory effects was the possible effect of MeOH-CNE, HaE-CNE on protein denaturation assay. Inflammation causes lysis of lysosomes which release their enzyme and produces a variety of diseases. NSAIDs exhibit their activity by inhibiting the release of lysosomal enzymes [<a href=\"#r-44\">44</a>]. Denaturation of proteins is initiated by inflammatory processes [<a href=\"#r-45\">45</a>]. Thus, NSAIDs provide protection against protein denaturation. So, the ability of MeOH-CNE, HaE-CNE to inhibit the protein denaturation may provide a significant contribution to its anti-inflammatory properties. The present result might be responsible for the presence of secondary metabolites such as alkaloid and flavonoid [<a href=\"#r-46\">46</a>].<br />\r\nThe available thrombolytic drugs in the market, particularly streptokinase, convert plasminogen to plasmin and increased clot lysis. According to the literature review explained that flavonoids, among the plant metabolites, affect thrombosis and cardiovascular disease by interfering with platelet activation. <em>Cocos nucifera </em>endocarp, especially HaE-CNE, showed a significant (<em>P <</em> 0.05) amount of clot lysis in comparison to the negative control. This result might be because of the presence of flavonoids which influence embolus and cardiovascular disorder by interfering with platelet actuation [<a href=\"#r-47\">47</a>].<br />\r\nBrine shrimp lethality assay is used for the cytotoxicity study. Ideally, the potential agent for the treatment of cancer should be nontoxic to a normal cell. However, anticancer agents are every so often lethal to normal cells, particularly towards rapidly growing cells. It is important to test this extract in low concentration to assess its potency [<a href=\"#r-48\">48</a>]. The results observed in 24 h were found to be nontoxic for extracts. Generally, the smaller the LC50, the higher the toxicity, and vice versa. The value of LC50 over 1000 µg/ml is considered to be nontoxic, ranging from 500 -1000 µg/ml is weakly toxic, moderately toxic for 100 – 500 µg/ml while less than 100 µg/ml is considered as highly toxic [<a href=\"#r-49\">49-69</a>]. No extracts were found to be toxic compared to Vincristine sulfate. The presence of alkaloids and saponins are present in this study, which can be effective as cytotoxic agents [<a href=\"#r-27\">27</a>]</p>\r\n\r\n<div id=\"figure6\">\r\n<figure class=\"image\"><img alt=\"\" height=\"248\" src=\"/media/article_images/2024/26/17/178-1579228061-Figure6.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 6. </strong>Graphical representations of pharmacological activities of <em>Cocos nucifera </em>endocarp.</figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 5,
"section_title": "CONCLUSIONS",
"body": "<p>Coconut endocarp extract can be regarded as a promising candidate with high therapeutic potential for drug preparation (Figure 6). The current study may provide useful data concerning the different medicinal properties in coconut shells for protecting humans against common diseases. From the above results, it may be concluded that the endocarp extract of <em>C. nucifera</em> exhibited secondary metabolites with a dose-dependent manner anti-inflammatory activity and moderate thrombolytic activity with lower cytotoxicity. Moreover, significant anxiolytic and antidiarrheal effects observed by both methanolic and hydro-alcoholic extract of<em> C. nucifera </em>endocarp. Therefore, further work, especially bioassay-guided fractionation, is warranted in order to isolate and characterize the active constituents responsible for the specific biological property.</p>"
},
{
"section_number": 6,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>Authors wish to thank the authority of International Islamic University Chittagong for their kind support in progress of the research. We also grateful to Laboratory of Alternative Medicine and Natural Products, Biochemistry and Molecular Biology, University of Chittagong for providing the facility to conducted this research. The authors are also thankful to Dr. Sheikh Bokhtear Uddin, Professor, Department of Botany, University of Chittagong, for identifying the plant.</p>"
},
{
"section_number": 7,
"section_title": "AUTHOR CONTRIBUTIONS",
"body": "<p>SA, MS, AMT, MSN, MAR, ASMAR, ZMB and MAH together planned and designed the research. ASMAR, MAR and TBE arranged the whole facilities for the research and supervised the whole research. SA, MS, AMT, MSN and MJR conducted the entire laboratory works with ZMB and MAH. MAR, ASMAR and TBE imparted in study design and interpreted the results putting efforts on statistical analysis and also participated in the manuscript draft and has thoroughly checked and revised the manuscript for necessary changes in format, grammar and English standard. All authors read and agreed on the final version of the manuscript.</p>"
},
{
"section_number": 8,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/26/17/178-1579228061-Figure1.jpg",
"caption": "Figure 1. Effect of MeOH-CNE and HaE-CNE on percentage of entry into open arm in elevated plus maze test in mice. Values are mean ± S.E.M. *P < 0.05 and **P < 0.01, significantly different from control; ANOVA followed Dunnett’s test (n = 6, per group). Where, MeOH-CNE: methanolic extract Cocos nucifera endocarp and HaE-CNE: hydro-alcoholic extract of Cocos nucifera endocarp.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/26/17/178-1579228061-Figure2.jpg",
"caption": "Figure 2. Antidiarrheal activity of MeOH-CNE and HaE-CNE against castor oil-induced diarrhea in mice. All values are expressed as mean ± SEM (n = 6); Data were analyzed by one way analysis of variance using GraphPad Prism for Windows, Version 6.0 (GraphPad software Inc., San Diego, CA, USA) followed by Dunnett’s test for multiple comparisons,. Values with * P < 0.05 were considered as significant. MeOH-CNE: methanolic extract Cocos nucifera endocarp and HaE-CNE: hydro-alcoholic extract of Cocos nucifera endocarp.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/26/17/178-1579228061-Figure3.jpg",
"caption": "Figure 3. Percentage inhibition of protein denaturation by MeOH-CNE, HaE-CNE. Results are mean ± SEM (n = 3). ANOVA followed by Dunnett’s test. Where, MeOH-CNE: methanolic extract Cocos nucifera endocarp and HaE-CNE: hydro-alcoholic extract of Cocos nucifera endocarp.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/26/17/178-1579228061-Figure4.jpg",
"caption": "Figure 4. Clot lysis by streptokinase, MeOH-CNE, HaE-CNE and water in vitro. Values are expressed as mean ± S.E.M. P < 0.05, significantly different from control; ANOVA followed by Dunnett’s test. Where, MeOH-CNE: methanolic extract Cocos nucifera endocarp and HaE-CNE: hydro-alcoholic extract of Cocos nucifera endocarp.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/26/17/178-1579228061-Figure5.jpg",
"caption": "Figure 5. Cytotoxic effects of MeOH-CNE and HaE-CNE. Extrapolation of inhibition concentrations of both MeOH-CNE, HaE-CNE and reference cytotoxic agent vincristine sulfate through regression analysis. Data are shown as mean ± SEM of 10 shrimps for each concentration. MeOH-CNE: methanolic extract Cocos nucifera endocarp and HaE-CNE: hydro-alcoholic extract of Cocos nucifera endocarp.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/26/17/178-1579228061-Figure6.jpg",
"caption": "Figure 6. Graphical representations of pharmacological activities of Cocos nucifera endocarp.",
"featured": false
}
],
"authors": [
{
"id": 832,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Saimun",
"family_name": "Akter",
"email": null,
"author_order": 1,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 833,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Masud",
"family_name": "Shah",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 834,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Abu Montakim",
"family_name": "Tareq",
"email": null,
"author_order": 3,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 835,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
},
{
"affiliation": "Department of Biochemistry & Molecular Biology, University of Chittagong, Chittagong-4331, Bangladesh"
}
],
"first_name": "Mst. Samima",
"family_name": "Nasrin",
"email": null,
"author_order": 4,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 836,
"affiliation": [
{
"affiliation": "Department of Biochemistry & Molecular Biology, University of Chittagong, Chittagong-4331, Bangladesh"
}
],
"first_name": "Md. Atiar",
"family_name": "Rahman",
"email": null,
"author_order": 5,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 837,
"affiliation": [
{
"affiliation": "International Institute for Halal Research and Training, KICT Building, Level 3, Jalan Gombak, 53100, Selangor, Malaysia"
}
],
"first_name": "Z. M.",
"family_name": "Babar",
"email": null,
"author_order": 6,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 838,
"affiliation": [
{
"affiliation": "Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan"
}
],
"first_name": "Md Anwarul",
"family_name": "Haque",
"email": null,
"author_order": 7,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 839,
"affiliation": [
{
"affiliation": "School of Life Science, University of Electronic Science and Technology of China, Sichuan, China"
}
],
"first_name": "Mohammad Jahir",
"family_name": "Royhan",
"email": null,
"author_order": 8,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 840,
"affiliation": [
{
"affiliation": "Department of Applied Chemistry and Chemical Engineering, Noakhali Science and Technology University, Sonapur, Noakhali-3814, Bangladesh"
}
],
"first_name": "Miskatun Nur",
"family_name": "Mamun",
"email": null,
"author_order": 9,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 190
},
{
"id": 841,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
},
{
"affiliation": "Department of Biochemistry & Molecular Biology, University of Chittagong, Chittagong-4331, Bangladesh"
}
],
"first_name": "A. S. M. Ali",
"family_name": "Reza",
"email": "alirezaru@gmail.com",
"author_order": 10,
"ORCID": null,
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "A.S.M. Ali Reza, Tel: +88-031-610085, E-mail: alirezaru@gmail.com",
"article": 190
},
{
"id": 842,
"affiliation": [
{
"affiliation": "Department of Pharmacy, BGC Trust University Bangladesh, Chittagong-4381, Bangladesh"
}
],
"first_name": "Talha Bin",
"family_name": "Emran",
"email": "talhabmb@gmail.com",
"author_order": 11,
"ORCID": "https://orcid.org/0000-0003-3188-2272",
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Talha Bin Emran, Cell: +88-01819942214, E-mail: talhabmb@gmail.com or talhabmb@bgctub.ac.bd",
"article": 190
}
],
"views": 1047,
"downloads": 105,
"references": [
{
"id": 6387,
"serial_number": 1,
"pmc": null,
"reference": "DebMandal M, Mandal S. Coconut (Cocos nucifera L.: Arecaceae): In health promotion and disease prevention2011.",
"DOI": null,
"article": 190
},
{
"id": 6388,
"serial_number": 2,
"pmc": null,
"reference": "Esquenazi D, Wigg MD, Miranda MM, Rodrigues HM, Tostes JB, Rozental S, da Silva AJ, Alviano CS. Antimicrobial and antiviral activities of polyphenolics from Cocos nucifera Linn. (Palmae) husk fiber extract. Res Microbiol. 2002;153:647-52.",
"DOI": null,
"article": 190
},
{
"id": 6389,
"serial_number": 3,
"pmc": null,
"reference": "Sachs M, Asskali F. Wound management with coconut oil in Indonesian folk medicine. Der Chirurg; Zeitschrift fur alle Gebiete der operativen Medizen. 2002;73:387-92.",
"DOI": null,
"article": 190
},
{
"id": 6390,
"serial_number": 4,
"pmc": null,
"reference": "Lima EBC, Sousa CNS, Meneses LN, Ximenes NC, Santos Júnior MA, Vasconcelos GS, Lima NBC, Patrocínio MCA, Macedo D, Vasconcelos SMM. Cocos nucifera (L.) (Arecaceae): A phytochemical and pharmacological review. Brazilian Journal of Medical and Biological Research. 2015;48:953-64.",
"DOI": null,
"article": 190
},
{
"id": 6391,
"serial_number": 5,
"pmc": null,
"reference": "Nagata JM, Jew AR, Kimeu JM, Salmen CR, Bukusi EA, Cohen CR. Medical pluralism on Mfangano Island: use of medicinal plants among persons living with HIV/AIDS in Suba District, Kenya. J Ethnopharmacol. 2011;135:501-9.",
"DOI": null,
"article": 190
},
{
"id": 6392,
"serial_number": 6,
"pmc": null,
"reference": "Al-Adhroey AH, Nor ZM, Al-Mekhlafi HM, Amran AA, Mahmud R. Evaluation of the use of Cocos nucifera as antimalarial remedy in Malaysian folk medicine. J Ethnopharmacol. 2011;134:988-91.",
"DOI": null,
"article": 190
},
{
"id": 6393,
"serial_number": 7,
"pmc": null,
"reference": "Emojevwe V. Cocos nucifera (Coconut) fruit: A review of its medical properties. 2013;3:718-23.",
"DOI": null,
"article": 190
},
{
"id": 6394,
"serial_number": 8,
"pmc": null,
"reference": "] Akinpelu DA, Alayande KA, Aiyegoro OA, Akinpelu OF, Okoh AI. Probable mechanisms of biocidal action of Cocos nucifera Husk extract and fractions on bacteria isolates. BMC Complement Altern Med. 2015;15:116.",
"DOI": null,
"article": 190
},
{
"id": 6395,
"serial_number": 9,
"pmc": null,
"reference": "Rinaldi S, Silva DO, Bello F, Alviano CS, Alviano DS, Matheus ME, Fernandes PD. Characterization of the antinociceptive and anti-inflammatory activities from Cocos nucifera L. (Palmae). J Ethnopharmacol. 2009;122:541-6.",
"DOI": null,
"article": 190
},
{
"id": 6396,
"serial_number": 10,
"pmc": null,
"reference": "Silva RR, Oliveira e Silva D, Fontes HR, Alviano CS, Fernandes PD, Alviano DS. Anti-inflammatory, antioxidant, and antimicrobial activities of Cocos nucifera var. typica. BMC Complement Altern Med. 2013;13:107.",
"DOI": null,
"article": 190
},
{
"id": 6397,
"serial_number": 11,
"pmc": null,
"reference": "Jose M, Sharma BB, Shantaram M, Ahmed SA. Ethnomedicinal herbs used in oral health and hygiene in coastal Dakshina Kannada. J Oral Health Comm Dent. 2011;5:119-23.",
"DOI": null,
"article": 190
},
{
"id": 6398,
"serial_number": 12,
"pmc": null,
"reference": "Jose M, Cyriac MB, Pai V, Varghese I, Shantaram M. Antimicrobial properties of Cocos nucifera (coconut) husk: An extrapolation to oral health. J Nat Sci Biol Med. 2014;5:359-64.",
"DOI": null,
"article": 190
},
{
"id": 6399,
"serial_number": 13,
"pmc": null,
"reference": "Kessler RC, Wang PS. The descriptive epidemiology of commonly occurring mental disorders in the United States. Annu Rev Public Health. 2008;29:115-29.",
"DOI": null,
"article": 190
},
{
"id": 6400,
"serial_number": 14,
"pmc": null,
"reference": "Hannah Ritchie, Roser M. Mental Health. Our World in Data. 2019.",
"DOI": null,
"article": 190
},
{
"id": 6401,
"serial_number": 15,
"pmc": null,
"reference": "Rex A, Schickert R, Fink H. Antidepressant-like effect of nicotinamide adenine dinucleotide in the forced swim test in rats. Pharmacology Biochemistry and Behavior. 2004;77:303-7.",
"DOI": null,
"article": 190
},
{
"id": 6402,
"serial_number": 16,
"pmc": null,
"reference": "Guzmán-Gutiérrez SL, Balderas J, Aguilar A, Navarrete A. Sedative activity of some plants used in Mexico to treat insomnia. Rev Latinoamer Quím. 2009;37:243-51.",
"DOI": null,
"article": 190
},
{
"id": 6403,
"serial_number": 17,
"pmc": null,
"reference": "Pawar VS, Anup A, Shrikrishna B, Shivakumar H. Antidepressant–like effects of Acorus calamus in forced swimming and tail suspension test in mice. Asian Pacific Journal of Tropical Biomedicine. 2011;1:S17-S9.",
"DOI": null,
"article": 190
},
{
"id": 6404,
"serial_number": 18,
"pmc": null,
"reference": "Shoba FG, Thomas M. Study of antidiarrhoeal activity of four medicinal plants in castor-oil induced diarrhoea. J Ethnopharmacol. 2001;76:73-6.",
"DOI": null,
"article": 190
},
{
"id": 6405,
"serial_number": 19,
"pmc": null,
"reference": "Guerrant RL, Van Gilder T, Steiner TS, Thielman NM, Slutsker L, Tauxe RV, Hennessy T, Griffin PM, DuPont H, Sack RB, Tarr P, Neill M, Nachamkin I, Reller LB, Osterholm MT, Bennish ML, Pickering LK. Practice guidelines for the management of infectious diarrhea. Clin Infect Dis. 2001;32:331-51.",
"DOI": null,
"article": 190
},
{
"id": 6406,
"serial_number": 20,
"pmc": null,
"reference": "Tenório JA, Dulciana S, da Silva TM, da Silva TG, Ramos CS. Solanum paniculatum root extract reduces diarrhea in rats. Revista Brasileira de Farmacognosia. 2016;26:375-8.",
"DOI": null,
"article": 190
},
{
"id": 6407,
"serial_number": 21,
"pmc": null,
"reference": "Hilal-Dandan R, Brunton L. Goodman and Gilman manual of pharmacology and therapeutics: McGraw Hill Professional; 2013.",
"DOI": null,
"article": 190
},
{
"id": 6408,
"serial_number": 22,
"pmc": null,
"reference": "Mekonnen B, Asrie AB, Wubneh ZB. Antidiarrheal activity of methanolic leaf extract of Justicia schimperiana. Evidence-Based Complementary and Alternative Medicine. 2018;2018:10.",
"DOI": null,
"article": 190
},
{
"id": 6409,
"serial_number": 23,
"pmc": null,
"reference": "Hanauer SB. The role of loperamide in gastrointestinal disorders. Rev Gastroenterol Disord. 2008;8:15-20.",
"DOI": null,
"article": 190
},
{
"id": 6410,
"serial_number": 24,
"pmc": null,
"reference": "Salama A, Levin Y, Jha P, Alweis R. Ventricular fibrillation due to overdose of loperamide, the “poor man’s methadone”. Journal of community hospital internal medicine perspectives. 2017;7:222-6.",
"DOI": null,
"article": 190
},
{
"id": 6411,
"serial_number": 25,
"pmc": null,
"reference": "Bankar GR, Nayak PG, Bansal P, Paul P, Pai KSR, Singla RK, Bhat VG. Vasorelaxant and antihypertensive effect of Cocos nucifera Linn. endocarp on isolated rat thoracic aorta and DOCA salt-induced hypertensive rats. Journal of ethnopharmacology. 2011;134:50-4.",
"DOI": null,
"article": 190
},
{
"id": 6412,
"serial_number": 26,
"pmc": null,
"reference": "Rodrigues S, Pinto GA. Ultrasound extraction of phenolic compounds from coconut (Cocos nucifera) shell powder. Journal of Food Engineering. 2007;80:869-72.",
"DOI": null,
"article": 190
},
{
"id": 6413,
"serial_number": 27,
"pmc": null,
"reference": "Tiwari P, Kumar B, Kaur M, Kaur G, Kaur HJIps. Phytochemical screening and extraction: a review. 2011;1:98-106.",
"DOI": null,
"article": 190
},
{
"id": 6414,
"serial_number": 28,
"pmc": null,
"reference": "Almeida RNdJEGK, Rio de Janeiro, Brasil. Psicofarmacologia: fundamentos práticos. 2006.",
"DOI": null,
"article": 190
},
{
"id": 6415,
"serial_number": 29,
"pmc": null,
"reference": "Costa de Melo N, Sánchez-Ortiz BL, Dos Santos Sampaio TI, Matias Pereira AC, Pinheiro da Silva Neto FL, Ribeiro da Silva H, Alves Soares Cruz R, Keita H, Soares Pereira AM, Tavares Carvalho JC. Anxiolytic and antidepressant effects of the hydroethanolic extract from the leaves of Aloysia polystachya (Griseb.) Moldenke: A study on zebrafish (Danio rerio). Pharmaceuticals (Basel, Switzerland). 2019;12:106.",
"DOI": null,
"article": 190
},
{
"id": 6416,
"serial_number": 30,
"pmc": null,
"reference": "Taufiq-Ur-Rahman M, Shilpi JA, Ahmed M, Hossain CF. Preliminary pharmacological studies on Piper chaba stem bark. Journal of Ethnopharmacology. 2005;99:203-9.",
"DOI": null,
"article": 190
},
{
"id": 6417,
"serial_number": 31,
"pmc": null,
"reference": "Bellah SF, Islam MN, Karim MR, Rahaman MM, Nasrin MS, Rahman MA, Reza AA. Evaluation of cytotoxic, analgesic, antidiarrheal and phytochemical properties of Hygrophila spinosa (T. Anders) whole plant. Journal of Basic and Clinical Physiology and Pharmacology. 2017;28:185-90.",
"DOI": null,
"article": 190
},
{
"id": 6418,
"serial_number": 32,
"pmc": null,
"reference": "Sakat S, Juvekar AR, Gambhire MN. In vitro antioxidant and anti-inflammatory activity of methanol extract of Oxalis corniculata Linn. Int J Pharm Pharm Sci. 2010;2:146-55.",
"DOI": null,
"article": 190
},
{
"id": 6419,
"serial_number": 33,
"pmc": null,
"reference": "Ansari P, Uddin MJ, Rahman MM, Abdullah-Al-Mamun M, Islam MR, Ali MH, Reza AA. Anti-inflammatory, anti-diarrheal, thrombolytic and cytotoxic activities of an ornamental medicinal plant: Persicaria orientalis. Journal of Basic and Clinical Physiology and Pharmacology. 2017;28:51-8.",
"DOI": null,
"article": 190
},
{
"id": 6420,
"serial_number": 34,
"pmc": null,
"reference": "Prasad S, Kashyap RS, Deopujari JY, Purohit HJ, Taori GM, Daginawala HF. Development of an in vitro model to study clot lysis activity of thrombolytic drugs. Thrombosis Journal. 2006;4:14.",
"DOI": null,
"article": 190
},
{
"id": 6421,
"serial_number": 35,
"pmc": null,
"reference": "Rahman MA, Sultana R, Emran TB, Islam MS, Rahman MA, Chakma JS, Rashid H-u, Hasan CMMJBc, medicine a. Effects of organic extracts of six Bangladeshi plants on in vitro thrombolysis and cytotoxicity. BMC Complement Altern Med. 2013;13:25.",
"DOI": null,
"article": 190
},
{
"id": 6422,
"serial_number": 36,
"pmc": null,
"reference": "Mandal S, Patra A, Samanta A, Roy S, Mandal A, Mahapatra TD, Pradhan S, Das K, Nandi DK. Analysis of phytochemical profile of Terminalia arjuna bark extract with antioxidative and antimicrobial properties. Asian Pacific journal of Tropical Biomedicine. 2013;3:960-6.",
"DOI": null,
"article": 190
},
{
"id": 6423,
"serial_number": 37,
"pmc": null,
"reference": "Ahmed S, Rakib A, Islam MA, Khanam BH, Faiz FB, Paul A, Chy MNU, Bhuiya NMA, Uddin MMN, Ullah SA. In vivo and in vitro pharmacological activities of Tacca integrifolia rhizome and investigation of possible lead compounds against breast cancer through in silico approaches. Clin Phytosci. 2019;5:36.",
"DOI": null,
"article": 190
},
{
"id": 6424,
"serial_number": 38,
"pmc": null,
"reference": "Griebel G, Belzung C, Perrault G, Sanger DJ. Differences in anxiety-related behaviours and in sensitivity to diazepam in inbred and outbred strains of mice. Psychopharmacology. 2000;148:164-70.",
"DOI": null,
"article": 190
},
{
"id": 6425,
"serial_number": 39,
"pmc": null,
"reference": "Patil V, V.M C, Hugar S. Evaluation of anti-anxiety activity of Cocos nucifera endocarp on anxiety models. Asian Journal of Pharmacy and Pharmacology. 2019;5.",
"DOI": null,
"article": 190
},
{
"id": 6426,
"serial_number": 40,
"pmc": null,
"reference": "Rizzo V, Clifford MN, Brown JE, Siracusa L, Muratore G. Effects of processing on the polyphenol and phenolic acid content and antioxidant capacity of semi‐dried cherry tomatoes (Lycopersicon esculentum M.). Journal of the Science of Food and Agriculture. 2016;96:2040-6.",
"DOI": null,
"article": 190
},
{
"id": 6427,
"serial_number": 41,
"pmc": null,
"reference": "Rahman MA, bin Imran T, Islam S. Antioxidative, antimicrobial and cytotoxic effects of the phenolics of Leea indica leaf extract. Saudi Journal of Biological Sciences. 2013;20:213-25.",
"DOI": null,
"article": 190
},
{
"id": 6428,
"serial_number": 42,
"pmc": null,
"reference": "Khalid S, AdilShahzad N, Muhammad A, Anwar P. Phytochemical screening and analysis of selected medicinal plants in Gujrat. Journal of Phytochemistry and Biochemistry. 2018;2:108.",
"DOI": null,
"article": 190
},
{
"id": 6429,
"serial_number": 43,
"pmc": null,
"reference": "Tiwari P, Kumar B, Kaur M, Kaur G, Kaur H. Phytochemical screening and extraction: A Review 2011.",
"DOI": null,
"article": 190
},
{
"id": 6430,
"serial_number": 44,
"pmc": null,
"reference": "Mounnissamy VM, Kavimani S, Balu V, Quine SD. Evaluation of anti-inflammatory and membrane stabilizing property of ethanol extract of Cansjera rheedii J. Gmelin (Opiliaceae). Iranian Journal of Pharmacology and Therapeutics. 2007;6:235-0.",
"DOI": null,
"article": 190
},
{
"id": 6431,
"serial_number": 45,
"pmc": null,
"reference": "Umapathy E, Ndebia E, Meeme A, Adam B, Menziwa P, Nkeh-Chungag B, Iputo J. An experimental evaluation of Albuca setosa aqueous extract on membrane stabilization, protein denaturation and white blood cell migration during acute inflammation. Journal of Medicinal Plants Research. 2010;4:789-95.",
"DOI": null,
"article": 190
},
{
"id": 6432,
"serial_number": 46,
"pmc": null,
"reference": "Shravan K, Kishore G, Siva K, Sindhu P. In vitro anti-inflammatory and anti-arthritic activity of leaves of Physalis angulata L. Int J Pharm Ind Res. 2011;1:211-3.",
"DOI": null,
"article": 190
},
{
"id": 6433,
"serial_number": 47,
"pmc": null,
"reference": "Dwivedi S. Terminalia arjuna Wight & Arn.—a useful drug for cardiovascular disorders. Journal of Ethnopharmacology. 2007;114:114-29.",
"DOI": null,
"article": 190
},
{
"id": 6434,
"serial_number": 48,
"pmc": null,
"reference": "Chowdhury TA, Kamal AM, Chowdhury KAA, Jahan A, Hossain MS, Mamur A, Hasan M, Hossain J. Cytotoxic & thrombolytic activity of methanolic extract of Macaranga denticulata Bark. The Pharma Innovation. 2015;4:36.",
"DOI": null,
"article": 190
},
{
"id": 6435,
"serial_number": 49,
"pmc": null,
"reference": "Nguta J, Mbaria J, Gakuya D, Gathumbi P, Kabasa J, Kiama S. Biological screening of Kenyan medicinal plants using Artemia salina (Artemiidae). Pharmacologyonline. 2011;2:458-78.",
"DOI": null,
"article": 190
},
{
"id": 6436,
"serial_number": 50,
"pmc": null,
"reference": "Rakib A, Ahmed S, Islam MA, Haye A, Uddin SMN, Uddin MMN, Hossain MK, Paul A, Emran, TB. Antipyretic and hepatoprotective potential of Tinospora crispa and investigation of possible lead compounds through in silico approaches. Food Sci Nutrition. 2020;8(1):547-556.",
"DOI": null,
"article": 190
},
{
"id": 6437,
"serial_number": 51,
"pmc": null,
"reference": "Shifah F, Tareq AM, Sayeed MA, Islam MN, Emran TB, Ullah MA, Mukit MA, Ullah M. Antidiarrheal, cytotoxic and thrombolytic activities of methanolic extract of Hedychium coccineum leaves. J Adv Biotechnol Exp Therapeutics. 2020;3(1):77-83.",
"DOI": null,
"article": 190
},
{
"id": 6438,
"serial_number": 52,
"pmc": null,
"reference": "Dutta T, Paul A, Majumder M, Sultan RA and Emran TB. Pharmacological evidence for the use of Cissus assamica as a medicinal plant in the management of pain and pyrexia. Biochem Biophy Rep. 2019;21:1-8.",
"DOI": null,
"article": 190
},
{
"id": 6439,
"serial_number": 53,
"pmc": null,
"reference": "Bulbul MRH, Rahman MA, Rahman MZ, Emran TB, Afroze M, Khan M, Chowdhury MAH, Ibrahim MA and Chowdhury MS (2019). Alcoholic extract of Leea macrophylla (Roxb.) root reverses CCl4 induced liver injury through upregulation of antioxidative enzyme’s gene mRNA expression: a molecular interaction for therapeutic inception. Oriental Pharmacy and Experimental Medicine. pp. 1-18.",
"DOI": null,
"article": 190
},
{
"id": 6440,
"serial_number": 54,
"pmc": null,
"reference": "Uddin MZ, Rana MS, Hossain S, Dutta E, Ferdous S, Dutta M and Emran TB (2019). In vivo neuroprotective, antinociceptive, anti-inflammatory potential in Swiss albino mice and in vitro antioxidant and clot lysis activities of fractionated Holigarna longifolia Roxb. bark extract. Journal of Complementary and Integrative Medicine. 17(2):1-9.",
"DOI": null,
"article": 190
},
{
"id": 6441,
"serial_number": 55,
"pmc": null,
"reference": "Emran TB, Dash R, Uddin MMN, Rahman MA. Sedative, anxiolytic, antinociceptive, anti-inflammatory and antipyretic effects of a chloroform extract from the leaves of Urena sinuata (Borss) L. in rodents. J Appl Life Sci Int. 2018;16(3): 1-19.",
"DOI": null,
"article": 190
},
{
"id": 6442,
"serial_number": 56,
"pmc": null,
"reference": "Uddin MMN, Ahmed S, Kabir MSH, Rahman MS, Sultan RA, Emran TB. (2017). In vivo analgesic, anti-inflammatory potential in Swiss albino mice and in vitro thrombolytic activity of hydroalcoholic fruits extract from Daemonorops robusta Warb. Journal of Applied Pharmaceutical Sciences. 7 (1): 104-113.",
"DOI": null,
"article": 190
},
{
"id": 6443,
"serial_number": 57,
"pmc": null,
"reference": "Al Mahmud Z, Qais N, Bachar SC, Hasan CM, Emran TB and Uddin MMN (2017). Phytochemical investigations and antioxidant potential of leaf of Leea macrophylla (Roxb.). BMC Res Notes. 2017;10: 245.",
"DOI": null,
"article": 190
},
{
"id": 6444,
"serial_number": 58,
"pmc": null,
"reference": "Al Mahmud Z, Emran TB, Qais N, Bachar SC, Sarker M and Uddin MMN (2016). Evaluation of analgesic, anti-inflammatory, thrombolytic and hepatoprotective activities of roots of Premna esculenta (Roxb). Journal of Basic and Clinical Physiology and Pharmacology. 27(1):63-70. DOI: 10.1515/jbcpp-2015-0056.",
"DOI": null,
"article": 190
},
{
"id": 6445,
"serial_number": 59,
"pmc": null,
"reference": "Dash R, Ahsan MT, Hosen SMZ, Rahman MG, Emran TB and Uddin MMN. Evolution of selective COX-2 inhibitor from Alangium salvifolium: an in silico approach. J Appl Pharm Sci. 2015;5(4):89-93.",
"DOI": null,
"article": 190
},
{
"id": 6446,
"serial_number": 60,
"pmc": null,
"reference": "Kabir MSH, Hossain MM, Kabir MI, Rahman MM, Hasanat A, Emran TB and Rahman MA. Phytochemical screening, antioxidant, thrombolytic, α-amylase inhibition and cytotoxic activities of ethanol extract of Steudnera colocasiifolia K. Koch leaves. J Young Pharmacists. 2016;8(4):391-397.",
"DOI": null,
"article": 190
},
{
"id": 6447,
"serial_number": 61,
"pmc": null,
"reference": "Uddin MMN, Zahan S, Islam MA, Ahmed S, Tajbiha-E-Mowla, Rahman MS, Sultan RA and Emran TB. Evaluation of the anti-diarrheal activity of methanol extract and its fractions of Urena sinuata L. (Borss) leaves. J Appl Pharm Sci. 2016;6(12):056-060.",
"DOI": null,
"article": 190
},
{
"id": 6448,
"serial_number": 62,
"pmc": null,
"reference": "Dash R, Emran TB, Paul A, Siddique MK, Khan MA, Rahman MG and Uddin MMN. (2016). Effects of five Bangladeshi plant extracts on in vitro thrombolysis and cytotoxicity. Pharmacognosy Research. 8(3):176-180.",
"DOI": null,
"article": 190
},
{
"id": 6449,
"serial_number": 63,
"pmc": null,
"reference": "Emran TB, Rahman MA, Uddin MMN, Rahman MM, Uddin MZ, Dash R and Layzu C. Effects of organic extracts and their different fractions of five Bangladeshi plants on in vitro thrombolysis. BMC Complement Altern Med. 2015;15:128.",
"DOI": null,
"article": 190
},
{
"id": 6450,
"serial_number": 64,
"pmc": null,
"reference": "Emran TB, Rahman MA, Uddin MMN, Dash R, Hossen MF, Mohiuddin M and Alam MR. Molecular docking and inhibition studies on the interactions of Bacopa monnieri’s potent phytochemicals against Staphylococcus aureus. DARU J Pharm Sci. 2015;23:26.",
"DOI": null,
"article": 190
},
{
"id": 6451,
"serial_number": 65,
"pmc": null,
"reference": "Rahman MA, Mahmud S, Akhter S, Aklima J, Akhter S, Merry SR, Jubair SMR, Dash R and Emran TB (2015). Anti-thrombotic effects of five organic extracts of Bangladeshi plants and in silico models for the mechanism of the observed effects. Evidence-Based Complementary and Alternative Medicine.",
"DOI": null,
"article": 190
},
{
"id": 6452,
"serial_number": 66,
"pmc": null,
"reference": "Dash R, Ahsan MT, Hosen SMZ, Rahman MG, Emran TB and Uddin MMN (2015). Evolution of selective COX-2 inhibitor from Alangium salvifolium: an in silico approach. Journal of Applied Pharmaceutical Sciences. 5(4): 89-93.",
"DOI": null,
"article": 190
},
{
"id": 6453,
"serial_number": 67,
"pmc": null,
"reference": "Momin MAM, Bellah SMF, Rahman SM, Rahman AA, Murshid GMM and Emran TB. Phytopharmacological evaluation of ethanol extract of Sida cordifolia L. roots. Asian Pac J Trop Biomed. 2014;4(1):18-24.",
"DOI": null,
"article": 190
},
{
"id": 6454,
"serial_number": 68,
"pmc": null,
"reference": "Biswas FB, Roy TG, Rahman MA, Emran TB. An in vitro antibacterial and antifungal effects of cadmium(II) complexes of hexamethyltetraazacyclotetradecadiene and isomers of its saturated analogue. Asian Pac J Trop Biomed. 2014;7(Suppl. 2):S534-S539.",
"DOI": null,
"article": 190
},
{
"id": 6455,
"serial_number": 69,
"pmc": null,
"reference": "Emran TB, Rahman MA (2014). Sedative, anxiolytic and analgesic effects of Urena sinuata L. leaf extract in animal models. International Food Research Journal. 21(5):2069-2075.",
"DOI": null,
"article": 190
}
]
},
{
"id": 186,
"slug": "178-1586622646-anxiolytic-and-thrombolytic-investigation-of-methanol-extract-of-piper-nigrum-l-fruits-and-sesamum-indicum-l-seeds",
"featured": false,
"slider": false,
"issue": "Vol3 Issue3",
"type": "original_article",
"manuscript_id": "178-1586622646",
"recieved": "2020-03-10",
"revised": null,
"accepted": "2020-04-22",
"published": "2020-06-01",
"pdf_file": "https://jabet.bsmiab.org/media/pdf_file/2023/44/178-1586622646.pdf",
"title": "Anxiolytic and thrombolytic investigation of methanol extract of Piper nigrum L. fruits and Sesamum indicum L. seeds",
"abstract": "<p>The study was intended to compare and evaluate the <em>in vitro</em> thrombolytic activity and <em>in vivo</em> anxiolytic activities of methanol extracts of <em>Piper nigrum</em> fruits (MEPNF) and <em>Sesamum indicum</em> seeds (MESIS). An <em>in vitro</em> thrombolytic activity was employed by investigating the clot lysis effect of MEPNF, MESIS, and their combinations. <em>In vivo</em> anxiolytic activity of MEPNF and MESIS was examined by using the Elevated Plus Maze (EPM) test considering diazepam (1 mg/kg) as the standard drug. In the thrombolytic test, MEPNF and MESIS showed a moderate (<em>P</em> < 0.01) clot lysis activity. Besides, MEPNF and MESIS showed significant (<em>P</em> < 0.001) dose-dependent anxiolytic activity. Since MEPNF or MESIS 200 and 400 (mg/kg, b.w; p.o) has been administered, the time spent in the open arms, and the number of entry into the open arms were increased. The finding suggests that MEPNF and MESIS may be the source for biological activity and therefore further research is required for confirmation.</p>",
"journal_reference": "J Adv Biotechnol Exp Ther. 2020; 3(3): 158-164.",
"academic_editor": "Dr. Md. Abdul Hannan, Dongguk University, South Korea.",
"cite_info": "Emon NU, Kaiser M, et al. Anxiolytic and thrombolytic investigation of methanol extract of Piper nigrum L. fruits and Sesamum indicum L. seeds. J Adv Biotechnol Exp Ther. 2020; 3(3): 158-164.",
"keywords": [
"Thrombolytic",
"Piper nigrum",
"Anxiolytic activity",
"Streptokinase",
"Sesamum indicum"
],
"DOI": "10.5455/jabet.2020.d121",
"sections": [
{
"section_number": 1,
"section_title": "INTRODUCTION",
"body": "<p>Cardiovascular disorders are one of the principal reasons beyond death instead of any other reason which is rising dramatically. The principal reason for the cardiovascular disease is because of thrombus (blood clot), which inhibits the blood circulation simply by constriction of the blood vessel and deprives the normal blood supply and oxygen to the tissue [<a href=\"#r-1\">1</a>]. Thrombolytic/fibrinolysis representatives are utilized to combat thrombosis and remove the clots [<a href=\"#r-2\">2</a>]. As tissue Urokinase (UK), streptokinase (SK), plasminogen activator (tPA), [<a href=\"#r-3\">3</a>] are frequently using a thrombolytic agent which used to treat fibrin or clot disorders. Therefore, specialists worldwide are using thrombolytic agents for the treatment of heart, and artery complications [<a href=\"#r-4\">4</a>]. Some thrombolytic substances have been obtained from several sources. Some of these thrombolytic agents are also assessed to indicate their direct and compelling development [<a href=\"#r-5\">5</a>]. Therefore, clot lysis is useful for clot-related disorders such as, deep vein thrombosis, thrombo-embolic strokes, myocardial infarction, and pulmonary embolism to eliminate a clogged artery that avoids permanent tissue damage [<a href=\"#r-6\">6</a>].<br />\r\nBesides, according to the World Health Report [<a href=\"#r-7\">7</a>], approximately 450 million people suffer from a mysterious or behavioral sickness, yet only a small minority of them receive even the most basic treatment. This amounts to 12.3% of the global burden of disease and will rise to 15% by 2020 [<a href=\"#r-8\">8</a>]. Anxiety is a persistent emotion, but when it occurs frequently then it turns into a horrific psychological condition. The presence of anxiety due to depression results in symptomatic complications, low accurate prognoses, worse medication response, and an increased risk of suicide [<a href=\"#r-9\">9</a>]. Worldwide medicinal plant work has progressed in the quest for alternative therapeutic agents for psychiatric disorders, the pharmacological effects of certain plants were demonstrated in many animal models. Medicinal plant products extracted from natural products are used extensively around the world to treat these chronic diseases and for the treatment of mild and life-threatening illness [<a href=\"#r-10\">10</a>]. Bangladeshi people living in rural areas consume medicinal plants as their principal source of health care and play a key role in treating many diseases [<a href=\"#r-11\">11</a>]. From that, point of view <em>Piper nigrum</em> (Family: Piperaceae) which is popularly called “Golmarich, Kalimeris” in West Bengal, India, and in Bangladesh. It is known as a hefty climbing bush. Leaves of this plant are extensively praised, whole, adjusted at the base, in a matter of seconds taper at zenith. <em>P.nigrum</em> contains alkaloids, aflatoxin, greasy oil oleoresin, and gum [<a href=\"#r-12\">12</a>].<em> </em>The fruit of the plant has been used as carminative, anti-periodic, rubefacient, stimulant, and aphrodisiac; used mostly for cough and cold recovery [<a href=\"#r-13\">13</a>].<br />\r\nAnother one is <em>Sesamum indicum</em> (Family: Edaliaceae) is commonly called “Sesame Benne”. Plants can be branched or un-branched. Seeds are remarkably rich in a fixed oil (up to 55%) comprising fundamentally of glycerides of oleic and linoleic acids and palmitic, stearic and myristic acids. Additionally, it also contains strong fats, stearin, palmitin, and myrisin, a crystalline substance, sesamin; sesamolin, which separates to a phenolic substance, sesamol, sesamin, protein, vitamin A and E, folic acid and minerals. Leaves contain a sticky substance and mucilage [<a href=\"#r-14\">14</a>]. A flavonoid glycoside – pedalin has been segregated from leaves [<a href=\"#r-13\">13</a>]. The plant has been traditionally used to treat clot lysis, high calories, ingestion, swelling, ulcers, asthma, and cough. The leaves and seeds are astringent. The combination of leaves with water was used to treat child cholera, diarrhea, dysentery, catarrh, and bladder disorders. [<a href=\"#r-15\">15</a>]. The existing research is to be investigated the thrombolytic and anxiolytic activities of methanol extracts of <em>Piper nigrum</em> fruits, <em>Sesamum indicum</em> seeds, and their combinations. Also, the study covers the anxiolytic activities of methanol extracts of <em>Piper nigrum</em> fruits and <em>Sesamum indicum</em> seeds.</p>"
},
{
"section_number": 2,
"section_title": "MATERIALS AND METHODS",
"body": "<p><strong>Collection and identification of the plants</strong><br />\r\nIn April 2019, <em>Piper nigrum </em>L. fruits and <em>Sesamum indicum </em>L<em>. </em>seeds were collected from the nearby forest of Chittagong, Bangladesh for the experiment. After that, the taxonomy of the plants has been confirmed by Dr. Sheikh Bokhtear Uddin, (Professor, Department of Botany, University of Chittagong, Chittagong-4331, Bangladesh) and stored the plant specimen for the further analysis (accession no: CTGUH-1335 and CTGUH-2712). After washing, the obtained plant fruits and seeds were dried throughout shade at low temperatures for twenty days and ground using a suitable blender machine. The coursed powder materials were then put in an impermeable container and kept in a cool, safe, and warm place until further study. In the case of ethical considerations, all the study protocols have been approved and the protocol was approved by the P&D committee (Pharm-P&D17/08’-19) of the department of pharmacy, International Islamic University Chittagong.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Animals</strong><br />\r\nFor the experiment, Swiss-albino mice (25-30 g) were obtained from the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B). Before the study, for one week, all animals were accustomed to the new environment. The animals were kept at 25 ° C temperature, comparative humidity (55-65%), and 12 h day/light cycle in a well-ventilated animal house during the experimental phase. Standard laboratory food and drinking water (<em>Ad libitum</em>) were given to the experimental animals.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Drug and chemicals</strong><br />\r\nThe drug that was exploited for the analyses of anxiolytic activity diazepam were purchased from Square Pharmaceutical Ltd, Bangladesh. For thrombolytic activity, economically available lyophilized Streptokinase (SK) vials (Polamin Werk GmbH, Herdecke, Germany) of 1500000 IU was borrowed from Sanofi Bangladesh Ltd. Besides, all other chemicals used in the experiment were pharmaceutical grade.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Extraction process</strong><br />\r\nThe granulated powder substance approximately (800 g) was soaked in 2.4 liters of methanol and kept in a glass bottle. The bottle was then shaken regularly and continued about 15 days for complete mixing. Then, it was filtered by the Whatman filter paper (Bibby RE200, Sterilin Ltd, UK) and concentrated by using a water bath at 60-65 °C temperature and finally, 11.27 g crude extract was obtained from <em>Piper nigrum</em> L. fruits and 8.2 g crude extract have been obtained S<em>esamum indicum</em> L. seeds. The concentrate was then kept at 4 °C for the reservation as well as for further investigation.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong><em>In vitro</em></strong><strong> thrombolytic test</strong><br />\r\n<em>Preparation of extract</em><br />\r\n10 mL of distilled water is used to suspend 10 mg of the extract and vigorously shaken by the vortex mixer. By that point, the suspension was vacuumed to remove the insoluble elements supernatant and was removed by filter paper (Whatman No. 1). The extracts were then prepared to evaluate the <em>in vitro</em> thrombolytic activity [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_4\">4</a>].</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>Streptokinase (SK) solution preparation</em><br />\r\n5 mL of clean distilled water was added to the commercially available SK-15, 00,000 I.U. (Polamin Werk GmbH) bottle that was then properly mixed. From the suspension, 100 μL (30,000 I.U) was used as a stock solution for the <em>in vitro</em> thrombolysis investigation. [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_4\">4</a>].</p>\r\n\r\n<p> </p>\r\n\r\n<p><em>Specimen for thrombolytic test</em><br />\r\nThe experiment was performed using the previously established protocol mentioned by Emon et.al; [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_16\">16</a>] and the study was approved by the Ethics Committee, Department of Pharmacy, International Islamic University Chittagong. For this research, blood (5 mL) were taken from the physically strong human volunteers (n = 10) having no history of NSAID and anticoagulants for 10 days. 500 μL of blood has been transferred to the early measured Eppendorf and sustained in the incubator at 37 °C for 45 minutes. After the formation of coagulation, the serum was completely separated from the Eppendorf. Each tube containing only coagulation has been weighed further in order to determine the exact weight of the coagulation. Then 100 μL of plant extract was added to the tubes. The test tube containing plant extract was suspended in the incubator and fixed the temperature at 37 ° C for 90 minutes. The blood serum has been withdrawn after the clot lysis and the tube has been measured once again to track the weight difference after the formation of clot lysis.<br />\r\n% clot lysis = Weight of the lysis clot / Weight of clot before lysis× 10</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Acute oral toxicity test</strong><br />\r\nA single oral dose limit of 2000 (mg/kg, b.w; p.o) for employing the acute oral toxicity test was carried out in compliance with the OECD 425 guidelines (OECD, 236) [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_17\">17</a>]. For this study, five swiss albino mice have randomly selected. One animal was first dosed at the target test dose and was then monitored for 24 hours. The remaining four animals were also dosed to 2000 mg/kg sequentially, for recording the overall testing of 5 animals. At least one in the first 30 minutes of dosage, regularly during the first 24 hours and daily for the next 13 days, for a total of 14 days, each animal was monitored individually for toxicity signs.</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong><em>In vivo</em></strong><strong> anxiolytic activity</strong><br />\r\n<em>Elevated plus maze test (EPM)</em><br />\r\nThe study for estimating anxiolytic activity has been employed by using the method which was listed in [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_18\">18</a>, <a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_19\">19</a>]. In an elevated plus-maze test, the instrument was formed with two open and two shut arms of wood over one another individually shaping a figure like plus-sign. The EPM (EPM; 30 cm × 6 cm, each arm) was arranged 50 cm over the ground. The mice were decorated into six groups where each group consisting of three mice. Group Ⅰ received control solutions (Tween-80, 10 mg/mL, b.w; p.o) and standard drug diazepam (1 mg/kg, b.w; i.p), has been administered to group Ⅱ. Besides, group Ⅲ-Ⅵ received MEPNF and MESIS (400 and 200 mg/kg; p.o) respectively. After the application of the medication, every animal was put at the focal point of the instrument to face one of the closed arms. The number of the entry in the open and close arm and time spent in the open arm has been recorded for 5 min after administration of the test samples [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_20\">20</a>].<br />\r\nPercentage of the period spent in the arm = Time in open arm/(Open arm time + close arm time)</p>\r\n\r\n<p> </p>\r\n\r\n<p><strong>Statistical analysis</strong><br />\r\nGraph pad prism version 5.0 was used to analyze experimental results. Data was submitted as SEM ± Mean. One-way ANOVA followed by Dunnett’s Multiple Comparison Test has performed for statistical analysis. The <em>P</em> < 0.05, <em>P</em> < 0.01, and <em>P</em> < 0.001 were considered statistically significant.</p>"
},
{
"section_number": 3,
"section_title": "RESULTS",
"body": "<p><strong>Effect of extracts on thrombolytic activity and acute toxicity</strong><br />\r\nThe expansion of 100 µL SK, a positive control (30,000 I.U.) to the coagulations alongside an hour and a half of brooding at 37 °C, showed 75.01 ± 3.20 % (<em>P</em> < 0.001) clot lysis. The <em>in vitro</em> thrombolytic action study uncovered that MEPNF and MESIS showed 35.4 ± 2.4% (<em>P </em>< 0.01) and 32.94 ± 1.23 % (P < 0.01) activity and the summary of the clot lysis of MEPNF and MESIS and their combination has been shown in <a href=\"#figure1\">Figure 1</a>. Further, no morbidity, skin deterioration, or any physical degradation has appeared as explained by acute toxicity.</p>\r\n\r\n<div id=\"figure1\">\r\n<figure class=\"image\"><img alt=\"\" height=\"526\" src=\"/media/article_images/2024/31/15/178-1586622646-Figure1.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 1.</strong><em><strong> </strong>In vitro </em>thrombolytic activity of test samples. Values are presented as mean ± SEM; One-way analysis of variance (ANOVA) followed by Dunnett’s test. *<em>P</em> <0.05, **<em>P</em> <0.01, and ***<em>P</em> < 0.001 is considered as significant compared with the control. Note: MEPNF = Methanol extract of <em>Piper nigrum</em> fruits and MESIS = Methanol extract <em>Sesamum indicum</em> seeds, SW = 1% Tween 80, and SKT = Streptokinase<strong>.</strong></figcaption>\r\n</figure>\r\n\r\n<p> </p>\r\n</div>\r\n\r\n<p><strong>Effect of extracts on elevated plus maze (EPM)</strong><br />\r\nIn the EPM, after the administration of test drugs, the number of open arm entries and spent time in the open arm has been increased significantly (<em>P </em>< 0.1, and <em>P</em> < 0.01). MEPNF and MESIS showed dose dependent reduction of the number of entrances in the close arm and the spent of time in the close arm. After the administration of MESIS 400 mg/kg, the mice showed maximum entrance (35.33 ± 1.85,<em> P</em> < 0.001) and spent comparatively high time (74.06 ± 2.51 sec,<em> P</em> < 0.001) in the open arm. The overall result has been shown in <a href=\"#figure2\">Figures 2</a> and <a href=\"#figure3\">3</a>.</p>\r\n\r\n<div id=\"figure2\">\r\n<figure class=\"image\"><img alt=\"\" height=\"500\" src=\"/media/article_images/2024/31/15/178-1586622646-Figure2.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 2. </strong><em>In vitro</em> thrombolytic activity (number of the entry in open arms) of test samples. Values are presented as mean ± SEM; One-way analysis of variance (ANOVA) followed by Dunnett’s test. **<em>P</em> <0.01, and ***<em>P</em> < 0.001 is considered as significant compared with the control. Note: MEPNF = Methanol extract of <em>Piper nigrum</em> fruits and MESIS = Methanol extract <em>Sesamum indicum</em> seeds, TWN = 1% Tween 80, and DFN = Diazepam<strong>.</strong></figcaption>\r\n</figure>\r\n</div>\r\n\r\n<div id=\"figure3\">\r\n<figure class=\"image\"><img alt=\"\" height=\"499\" src=\"/media/article_images/2024/31/15/178-1586622646-Figure3.jpg\" width=\"500\" />\r\n<figcaption><strong>Figure 3.</strong> <em>In vivo</em> anxiolytic activity (time spent in open arms) of test samples. Values are presented as mean ± SEM; One-way analysis of variance (ANOVA) followed by Dunnett’s test. ***<em>P</em> < 0.001 is considered as significant compared with the control. Note: MEPNF = Methanol extract of <em>Piper nigrum</em> fruits and MESIS = Methanol extract <em>Sesamum indicum</em> seeds, TWN = 1% Tween 80, and DFN = Diazepam<strong>.</strong></figcaption>\r\n</figure>\r\n</div>"
},
{
"section_number": 4,
"section_title": "DISCUSSION",
"body": "<p>Phytopharmaceutical work has now developed a new field for the discovery of drugs from plant derivate that cure certain disorders and renew the focus in herbal medicines. Around 30 % of medicines are made from plant derivatives [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_21\">21</a>]. This study examined the thrombolytic and anxiolytic effect of methanol extract of <em>Piper nigrum </em>fruits (MEPNF), methanol extract of <em>Sesamum indicum </em>seeds (MESIS), and their combination<em>. </em>Several researchers have conducted a series of studies to identify plants and natural food resources which contains thrombolytic (anti-platelet, and anti-coagulant) activity and the finding suggests that the intake of such a food contributes to stokes and cardiac disorder to prevention [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_22\">22</a>]. There are several plant products with a thrombolytic capacity. These are Garlic (<em>Allium sativum</em>) [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_23\">23</a>], <em>Flammulina velutipes</em> [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_24\">24</a>], <em>Pleurotus ostreatus</em> [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_25\">25</a>], <em>Crocus sativus</em> Linn (indraceae) [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_4\">4</a>], Ginger (<em>Zingiber officinale</em>) [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_26\">26</a>], <em>Lumbricus rubellus</em> [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_27\">27</a>], and <em>Ganoderma lucidum</em> [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_28\">28</a>], chungkook-jang [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_29\">29</a>], <em>Spirodela polyrhiza</em> [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_28\">28</a>], and natto [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_30\">30</a>] correspondingly. Our analysis of MEPNF, MESIS, and MEPNF + MESIS indicates a moderate probability of clot lysis as opposed to saline water. Compared to the controls, the rise in clotting lysis by MEPNF, MESIS, and MEPNF+ MESIS indicates its potential application in coagulation-related disorders. The plasminogen enzyme stimulates the majority of thrombolytic agents and removes the related fibrin in order to restore blood supply into blood congested vessels [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_31\">31</a>]. The clot lysis activity was observed both for the MEPNF and MESIS, which mean methanol soluble compounds of <em>Piper nigrum</em> fruits and <em>Sesamum indicum</em> seeds contain thrombolytic responsible constituents [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_32\">32</a>]. The plasminogen cell-binding surface is readily stimulated through plasmin that can result in fibrinolysis whereas other plants are using their enzymes as a fibrinolysis effect [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_16\">16</a>]. Furthermore, the plasminogen activator bacterial, Staphylokinase streptokinase, serves as cofactor molecules that help to form exosites and enhance the enzyme’s substrate presentation. Staphylokinase stimulates plasminogen to dissolve the blood clotting and also destroys cellular extra-cellular matrix and fibrin fibers [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_33\">33</a>].<br />\r\nThe EPM is a widely recognized assessment and is particularly susceptible to the actions of both anxiolytic and anxiogenic medications on type A (GABAA) gamma amino-butyric corrosive benzodiazepine complex [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_34\">34</a>]. In EPM, normally standard mice would recommend passing much of their time in the closed arms. That preference seems to reflect an aversion to open arms generated by open space fears. A drug such as diazepam that improves open arm exploration is considered anxiolytic and the reverse is true for anxiogenic substances [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_35\">35</a>]. The methanol extract of <em>Piper nigrum</em> fruits and <em>Sesamum indicum</em> seeds (200 and 400 mg/kg b.w) has had an anxiolytic effect in mice because it increases open arm entry and spends more time in open arms of the EPM. These results may be linked to the effects of extract components that have on the behavior of γ-aminobutyric acid transaminase (GABA-t). They are supposed to increase the levels of GABA in the brain to suppress anxiety [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_36\">36</a>]. Acute therapy of methanol extract or dried leaves has been shown to improve calmness, modulate moods, and alleviate experimental tension in healthy young participants [<a href=\"https://www.bsmiab.org/jabet/anxiolytic-thrombolytic-investigation-methanol-extract-piper-nigrum-l-fruits-sesamum-indicum-l-seeds/#_ENREF_37\">37</a>]. In our study, the MEPNF, MESIS, and MEPNF+MESIS showed moderate thrombolytic activity which supposed to their phytochemicals. Besides, MEPNF and MESIS showed significant anxiolytic activity in animal model which may be possible because of the presence of phytochemicals that increase the level of GABA in the brain.</p>"
},
{
"section_number": 5,
"section_title": "CONCLUSIONS",
"body": "<p>This study tends to be presumed that, the methanol extracts of <em>Piper nigrum </em>fruits and <em>Sesamum indicum</em> seeds have significant possibilities as a candidate for anxiolytic activity having moderate thrombolytic properties and fascinated to discover the dynamic component(s) in charge of the anxiolytic and thrombolytic effect of these plants. This is just an initial report so that, the author suggests to investigate in further to identify the medicinal and medical possibilities.</p>"
},
{
"section_number": 6,
"section_title": "ACKNOWLEDGEMENT",
"body": "<p>The researcher would like to thank Mr. A.T.M. Mostafa Kamal, chairman and Assistant Professor, and the Department of Pharmacy, International Islamic University Chittagong, Chittagong 4318, Bangladesh for their supports and facilities for carrying out this research work.</p>"
},
{
"section_number": 7,
"section_title": "AUTHORS CONTRIBUTIONS",
"body": "<p>The research protocol was conceptualized, prepared, and designed by NUE. SA and MNI supervised the investigations. Along with NUE the laboratory experiment performed by MJU, MFIK, MAJ, SMT, ANMR. Together, MI and MK collected and analyzed the information. Data processing and software analysis were performed by NUE and MI. In preparation of the draft manuscript, NUE and MK participated. SA and MNI reviewed and corrected the error of the draft and with NUE they finalized the draft. A native English speaker then reviewed the text, and all the authors agreed eventually to publish the manuscript draft.</p>"
},
{
"section_number": 8,
"section_title": "FUNDING",
"body": "<p>The study was conducted by the self-financial means.</p>"
},
{
"section_number": 9,
"section_title": "CONFLICTS OF INTEREST",
"body": "<p>Authors declared that they have no conflict of interest.</p>"
}
],
"figures": [
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/31/15/178-1586622646-Figure1.jpg",
"caption": "Figure 1. In vitro thrombolytic activity of test samples. Values are presented as mean ± SEM; One-way analysis of variance (ANOVA) followed by Dunnett’s test. *P <0.05, **P <0.01, and ***P < 0.001 is considered as significant compared with the control. Note: MEPNF = Methanol extract of Piper nigrum fruits and MESIS = Methanol extract Sesamum indicum seeds, SW = 1% Tween 80, and SKT = Streptokinase.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/31/15/178-1586622646-Figure2.jpg",
"caption": "Figure 2. In vitro thrombolytic activity (number of the entry in open arms) of test samples. Values are presented as mean ± SEM; One-way analysis of variance (ANOVA) followed by Dunnett’s test. **P <0.01, and ***P < 0.001 is considered as significant compared with the control. Note: MEPNF = Methanol extract of Piper nigrum fruits and MESIS = Methanol extract Sesamum indicum seeds, TWN = 1% Tween 80, and DFN = Diazepam.",
"featured": false
},
{
"figure": "https://jabet.bsmiab.org/media/article_images/2024/31/15/178-1586622646-Figure3.jpg",
"caption": "Figure 3. In vivo anxiolytic activity (time spent in open arms) of test samples. Values are presented as mean ± SEM; One-way analysis of variance (ANOVA) followed by Dunnett’s test. ***P < 0.001 is considered as significant compared with the control. Note: MEPNF = Methanol extract of Piper nigrum fruits and MESIS = Methanol extract Sesamum indicum seeds, TWN = 1% Tween 80, and DFN = Diazepam.",
"featured": false
}
],
"authors": [
{
"id": 802,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Nazim Uddin",
"family_name": "Emon",
"email": "nazim7emon@gmail.com",
"author_order": 1,
"ORCID": "https://orcid.org/0000-0001-7567-4796",
"corresponding": true,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "Nazim Uddin Emon, Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh, Phone:+8801648094589, E-mail: nazim7emon@gmail.com",
"article": 186
},
{
"id": 803,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Md",
"family_name": "Kaiser",
"email": null,
"author_order": 2,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 804,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Mohidol",
"family_name": "Islam",
"email": null,
"author_order": 3,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 805,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Mohammed Faisal Ibna",
"family_name": "Kabir",
"email": null,
"author_order": 4,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 806,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Mohammad Jamir",
"family_name": "Uddin",
"email": null,
"author_order": 5,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 807,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Mifta Ahmed",
"family_name": "Jyoti",
"email": null,
"author_order": 6,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 808,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Saifullah Mansur",
"family_name": "Tanjil",
"email": null,
"author_order": 7,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 809,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Abu Nayem Mohammad",
"family_name": "Rasel",
"email": null,
"author_order": 8,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 810,
"affiliation": [
{
"affiliation": "Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Dhaka, Dhaka-1000, Bangladesh."
}
],
"first_name": "Safaet",
"family_name": "Alam",
"email": null,
"author_order": 9,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
},
{
"id": 811,
"affiliation": [
{
"affiliation": "Department of Pharmacy, International Islamic University Chittagong, Chittagong-4318, Bangladesh"
}
],
"first_name": "Mohammad Nazmul",
"family_name": "Islam",
"email": null,
"author_order": 10,
"ORCID": null,
"corresponding": false,
"co_first_author": false,
"co_author": false,
"corresponding_author_info": "",
"article": 186
}
],
"views": 660,
"downloads": 105,
"references": [
{
"id": 6202,
"serial_number": 1,
"pmc": null,
"reference": "Sai Sandeep Y, Panigrahi M, Divya G, Beena D. Evaluation of in vitro thrombolytic activity of phytochemicals in Bacopa monnieri Linn. Journal of Pharmacy Research. 2012;5:100-1.",
"DOI": null,
"article": 186
},
{
"id": 6203,
"serial_number": 2,
"pmc": null,
"reference": "Apu AS, Chowdhury FA, Khatun F, Jamaluddin A, Pathan AH, Pal A. Phytochemical screening and in vitro evaluation of pharmacological activities of Aphanamixis polystachya (Wall) Parker fruit extracts. Tropical Journal of Pharmaceutical Research. 2013;12:111-6.",
"DOI": null,
"article": 186
},
{
"id": 6204,
"serial_number": 3,
"pmc": null,
"reference": "Mucklow J. Thrombolytic treatment. Streptokinase is more economical than alteplase. BMJ: British Medical Journal. 1995;311:1506.",
"DOI": null,
"article": 186
},
{
"id": 6205,
"serial_number": 4,
"pmc": null,
"reference": "Prasad S, Kashyap RS, Deopujari JY, Purohit HJ, Taori GM, Daginawala HF. Effect of Fagonia arabica (Dhamasa) on in vitro thrombolysis. BMC Complementary and Alternative Medicine. 2007;7:36.",
"DOI": null,
"article": 186
},
{
"id": 6206,
"serial_number": 5,
"pmc": null,
"reference": "Komorowicz E, Kolev K, Lerant I, Machovich R. Flow rate–modulated dissolution of fibrin with clot-embedded and circulating proteases. Circulation research. 1998;82:1102-8.",
"DOI": null,
"article": 186
},
{
"id": 6207,
"serial_number": 6,
"pmc": null,
"reference": "Rahman MA, Sultana R, Emran TB, Islam MS, Rahman MA, Chakma JS, et al. Effects of organic extracts of six Bangladeshi plants on in vitro thrombolysis and cytotoxicity. BMC Complement Altern Med. 2013;13:25.",
"DOI": null,
"article": 186
},
{
"id": 6208,
"serial_number": 7,
"pmc": null,
"reference": "Organization WH. The World Health Report 2001: Mental health: new understanding, new hope: World Health Organization; 2001.",
"DOI": null,
"article": 186
},
{
"id": 6209,
"serial_number": 8,
"pmc": null,
"reference": "Herrera-Ruiz M, García-Beltrán Y, Mora S, Díaz-Véliz G, Viana GS, Tortoriello J, et al. Antidepressant and anxiolytic effects of hydroalcoholic extract from Salvia elegans. J Ethnopharmaco. 2006;107:53-8.",
"DOI": null,
"article": 186
},
{
"id": 6210,
"serial_number": 9,
"pmc": null,
"reference": "Kara S, Yazici KM, Güleç C, Ünsal I. Mixed anxiety–depressive disorder and major depressive disorder: comparison of the severity of illness and biological variables. Psychiatry research. 2000;94:59-66.",
"DOI": null,
"article": 186
},
{
"id": 6211,
"serial_number": 10,
"pmc": null,
"reference": "Adnan M, Chy NU, Mostafa Kamal A, Azad MOK, Paul A, Uddin SB, et al. Investigation of the biological activities and characterization of bioactive constituents of Ophiorrhiza rugosa var. prostrata (D. Don) & Mondal leaves through in vivo, in vitro, and in silico approaches. Molecules (Basel, Switzerland). 2019;24:1367.",
"DOI": null,
"article": 186
},
{
"id": 6212,
"serial_number": 11,
"pmc": null,
"reference": "Mollik MAH, Hossan MS, Paul AK, Taufiq-Ur-Rahman M, Jahan R, Rahmatullah M. A comparative analysis of medicinal plants used by folk medicinal healers in three districts of Bangladesh and inquiry as to mode of selection of medicinal plants. Ethnobotany Research and Applications. 2010;8:195-218.",
"DOI": null,
"article": 186
},
{
"id": 6213,
"serial_number": 12,
"pmc": null,
"reference": "Rastogi RM. Compendium of Indian medicinal plants. Central Drug Research Institute, Lucknow, India. 1990;1:388-9.",
"DOI": null,
"article": 186
},
{
"id": 6214,
"serial_number": 13,
"pmc": null,
"reference": "Rastogi RP, Mehrotra B. Compendium of Indian medicinal plants: Central Drug Research Institute; 1990.",
"DOI": null,
"article": 186
},
{
"id": 6215,
"serial_number": 14,
"pmc": null,
"reference": "Hasan SR, Hossain MM, Akter R, Jamila M, Mazumder MEH, Rahman S. DPPH free radical scavenging activity of some Bangladeshi medicinal plants. Journal of Medicinal plants research. 2009;3:875-9.",
"DOI": null,
"article": 186
},
{
"id": 6216,
"serial_number": 15,
"pmc": null,
"reference": "Ibrahim M, Tarrannum T, Ahsan Q, Kuddus MR, Rashid RB, Rashid MA. Anti-diarrheal, antimicrobial and membrane stabilizing activity of sarcochlamys pulcherrima gaudich. 2014.",
"DOI": null,
"article": 186
},
{
"id": 6217,
"serial_number": 16,
"pmc": null,
"reference": "Emon NU, Jahan I, Sayeed MA. Investigation of antinociceptive, anti-inflammatory and thrombolytic activity of Caesalpinia digyna (Rottl.) leaves by experimental and computational approaches. Advances in Traditional Medicine. 2020:1-9.",
"DOI": null,
"article": 186
},
{
"id": 6218,
"serial_number": 17,
"pmc": null,
"reference": "Kifayatullah M, Mustafa MS, Sengupta P, Sarker MMR, Das A, Das SK. Evaluation of the acute and sub-acute toxicity of the ethanolic extract of Pericampylus glaucus (Lam.) Merr. in BALB/c mice. Journal of Acute Disease. 2015;4:309-15.",
"DOI": null,
"article": 186
},
{
"id": 6219,
"serial_number": 18,
"pmc": null,
"reference": "Lister RG. The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology. 1987;92:180-5.",
"DOI": null,
"article": 186
},
{
"id": 6220,
"serial_number": 19,
"pmc": null,
"reference": "Pellow S, Chopin P, File SE, Briley M. Validation of open: closed arm entries in an elevated plus-maze as a measure of anxiety in the rat. Journal of neuroscience methods. 1985;14:149-67.",
"DOI": null,
"article": 186
},
{
"id": 6221,
"serial_number": 20,
"pmc": null,
"reference": "Ali SF, Chandra O, Hasan M. Effects of an organophosphate (dichlorvos) on open field behavior and locomotor activity: correlation with regional brain monoamine levels. Psychopharmacology. 1980;68:37-42.",
"DOI": null,
"article": 186
},
{
"id": 6222,
"serial_number": 21,
"pmc": null,
"reference": "Ramjan A, Hossain M, Runa JF, Md H, Mahmodul I. Evaluation of thrombolytic potential of three medicinal plants available in Bangladesh, as a potent source of thrombolytic compounds. Avicenna journal of phytomedicine. 2014;4:430.",
"DOI": null,
"article": 186
},
{
"id": 6223,
"serial_number": 22,
"pmc": null,
"reference": "Emon NU, Jahan I, Rudra S, Aktar M, Islam N. In Vitro Comparative Cardioprotective Activity of Methanol Extract of Caesalpinia Digyna (Rottl.) Stems and Senna Sophera (L.) Roxb. Stems.",
"DOI": null,
"article": 186
},
{
"id": 6224,
"serial_number": 23,
"pmc": null,
"reference": "Bordia A, Verma S, Srivastava K. Effect of garlic (Allium sativum) on blood lipids, blood sugar, fibrinogen and fibrinolytic activity in patients with coronary artery disease. Prostaglandins, leukotrienes and essential fatty acids. 1998;58:257-63.",
"DOI": null,
"article": 186
},
{
"id": 6225,
"serial_number": 24,
"pmc": null,
"reference": "Shin HH, Choi HS. Purification and partial characterization of a metalloprotease in Flammulina velutipes. Journal of Microbiology-Seoul-. 1998;36:20-5.",
"DOI": null,
"article": 186
},
{
"id": 6226,
"serial_number": 25,
"pmc": null,
"reference": "Choi H-S, Shin H-H. Purification and partial characterization of a fibrinolytic protease in Pleurotus ostreatus. Mycologia. 1998;90:674-9.",
"DOI": null,
"article": 186
},
{
"id": 6227,
"serial_number": 26,
"pmc": null,
"reference": "Verma S, Bordia A. Ginger, fat and fibrinolysis. Indian journal of medical sciences. 2001;55:83-6.",
"DOI": null,
"article": 186
},
{
"id": 6228,
"serial_number": 27,
"pmc": null,
"reference": "Jeon O-H, Moon W-J, Kim D-S. An anticoagulant/fibrinolytic protease from Lumbricus rubellus. BMB Reports. 1995;28:138-42.",
"DOI": null,
"article": 186
},
{
"id": 6229,
"serial_number": 28,
"pmc": null,
"reference": "Choi H-S, Sa Y-S. Fibrinolytic and antithrombotic protease from Ganoderma lucidum. Mycologia. 2000;92:545-52.",
"DOI": null,
"article": 186
},
{
"id": 6230,
"serial_number": 29,
"pmc": null,
"reference": "Kim W, Choi K, Kim Y, Park H, Choi J, Lee Y, et al. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from Chungkook-Jang. Appl Environ Microbiol. 1996;62:2482-8.",
"DOI": null,
"article": 186
},
{
"id": 6231,
"serial_number": 30,
"pmc": null,
"reference": "Sumi H, Hamada H, Tsushima H, Mihara H, Muraki H. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet. Experientia. 1987;43:1110-1.",
"DOI": null,
"article": 186
},
{
"id": 6232,
"serial_number": 31,
"pmc": null,
"reference": "Bhattacharya S, Ploplis VA, Castellino FJ. Bacterial plasminogen receptors utilize host plasminogen system for effective invasion and dissemination. BioMed Research International. 2012;2012.",
"DOI": null,
"article": 186
},
{
"id": 6233,
"serial_number": 32,
"pmc": null,
"reference": "Emran TB, Rahman MA, Uddin MMN, Rahman MM, Uddin MZ, Dash R, et al. Effects of organic extracts and their different fractions of five Bangladeshi plants on in vitro thrombolysis. BMC Complement Altern Med. 2015;15:128.",
"DOI": null,
"article": 186
},
{
"id": 6234,
"serial_number": 33,
"pmc": null,
"reference": "Parry MA, Zhang XC, Bode W. Molecular mechanisms of plasminogen activation: bacterial cofactors provide clues. Trends Biochem Sci. 2000;25:53-9.",
"DOI": null,
"article": 186
},
{
"id": 6235,
"serial_number": 34,
"pmc": null,
"reference": "Thakur V, Mengi S. Neuropharmacological profile of Eclipta alba (Linn.) Hassk. Journal of Ethnopharmacology. 2005;102:23-31.",
"DOI": null,
"article": 186
},
{
"id": 6236,
"serial_number": 35,
"pmc": null,
"reference": "Hasan M, Motin M. New slotting technique of making compact octagonal patch for four band applications. International Journal of Innovation and Applied Studies. 2013;3:221-7.",
"DOI": null,
"article": 186
},
{
"id": 6237,
"serial_number": 36,
"pmc": null,
"reference": "Ibarra A, Feuillere N, Roller M, Lesburgere E, Beracochea D. Effects of chronic administration of Melissa officinalis L. extract on anxiety-like reactivity and on circadian and exploratory activities in mice. Phytomedicine. 2010;17:397-403.",
"DOI": null,
"article": 186
},
{
"id": 6238,
"serial_number": 37,
"pmc": null,
"reference": "Kennedy DO, Little W, Haskell CF, Scholey AB. Anxiolytic effects of a combination of Melissa ofcinalis and Valeriana ofcinalis during laboratory induced stress. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2006;20:96-102.",
"DOI": null,
"article": 186
}
]
}
]
}
