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Marine Fisheries : J. of Marine Fisheries Technology and Management     Open Access   (Followers: 3)
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HAYATI Journal of Biosciences
Number of Followers: 0  

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ISSN (Print) 1978-3019 - ISSN (Online) 2086-4094
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  • Genipin Crosslinked Blended Collagen-Chondroitin: A Promising Biomaterial
           Scaffold Candidate for Cartilage Reconstruction

    • Authors: Trimartani Trimartani, Normalina Sandora, Bambang Hermani, Jeanne A Pawitan, Raden Ayu Anatriera
      Pages: 122 - 128
      Abstract: Tissue engineering offers a solution to the shortage of materials for cartilage reconstruction surgery by providing various potential biomaterial scaffolds. Tissue engineering utilizes biological or synthetic biomaterials as a scaffold for the host cells to repopulate and regenerate the tissue. The natural biomaterials such collagen and chondroitin imitates native cartilage matrix composition. Genipin as one of natural crosslinkers was added to improve the matrix biomechanical properties. This study was done to investigate biocomposition of blended collagen type 1, collagen type 2, chondroitin sulphate (Col1-Col2-CS) and genipin for its cytotoxicity using human umbilical mesenchymal stem cells (hUCMSCs), surface morphology, and biochemical composition. Genipin-crosslinked collagen-chondroitin biocomposite showed a homogeneous shape while uncrosslinked biocomposite had rough surface and fibrillar folds size. Spectroscopy demonstrated both biocomposites had similar peak resemble to no alternation of the biocomposition by crosslinking. Both types of biocomposites were biocompatible and had no toxic effects, as compared to the cell colony only (p value = 0.26). The conclusion are blended composite of collagen chondroitin crosslinked with genipin had generated a fine microstructure scaffold with smaller pore size, had similar biomolecular component spectrum absorption, and no exhibition of residual toxicity.
      PubDate: 2022-01-03
      DOI: 10.4308/hjb.29.2.122-128
      Issue No: Vol. 29, No. 2 (2022)
  • Genetic Diversity and Population Structure of Canarium tramdenum Dai and
           Yakovl. in Northern Vietnam

    • Authors: Phi Hong Hai, Nguyen Thi Huyen, Ha Thi Huyen Ngoc, Tran Thi Thu Ha, Nguyen Thi Viet Ha, La Anh Duong, Pham Huu Thuong, Alice Muchugi, Le Son
      Pages: 129 - 136
      Abstract: Canarium tramdenum occurs naturally in subtropical and tropical regions of Indochina and China. The wood is used for making high quality furniture and the fruit and leaves are used in traditional medicine. However, a lack of information on genetic diversity and population structure has handicapped the genetic conservation and domestication of this high-value species. This study evaluated genetic variation within and among four C. tramdenum populations. Sixty individuals were collected from four natural populations in Vietnam in the provinces of Ninhbinh, Bacgiang, Nghean, and Backan. Genetic diversity and genetic structure were determined using 20 ISSR markers. A total of 192 DNA fragments with sizes ranging from 110 bp to 3,000 bp were detected, of which 154 segments (80.2%) were polymorphic and 38 segments (19.8%) were monomorphic. The ISSR data indicated a moderate degree of genetic diversity for the species (h = 0.252). The four populations were separated into three genetic clusters with low levels of genetic distance between them. AMOVA result showed that most (78%) of the genetic variation was within the populations. The moderate to high genetic diversity of C. tramdenum and the low genetic differentiation among populations suggested that all existing natural populations in the particular regions needed to be preserved to protect the genetic diversity of this species.
      PubDate: 2022-01-03
      DOI: 10.4308/hjb.29.2.129-136
      Issue No: Vol. 29, No. 2 (2022)
  • Aeromonas hydrophila AHL 0905-2 and Streptococcus agalactiae N14G as
           Combined Vaccine Candidates for Nile Tilapia

    • Authors: Nunak Nafiqoh, Hessy Novita, Desy Sugiani, Lila Gardenia, Taukhid Taukhid, Arynta Widyaningrum, Dine Resti Susanti
      Pages: 137 - 145
      Abstract: In Indonesia, the Nile tilapia (Oreochromis niloticus) is the most widely farmed and available fish for consumption. Production loss due to bacterial infection by Aeromonas hydrophila and Streptococcus agalactiae is the main problem in tilapia cultivation. This study aimed to determine the occurrence of concurrent infection of Aeromonas hydrophila AHL 0905-2 and Streptococcus agalactiae N14G in Nile tilapia based on biochemical and molecular characteristics. From the results of biochemical assay and sequence analysis of the 16S rRNA fragment, Aeromonas hydrophila and Streptococcus agalactiae were confirmed. Genes for aerolysin (417 bp), nuclease (504 bp), lipase (155 bp), and serine protease (211 bp) were found in Aeromonas hydrophila AHL 0905-2, while Streptococcus agalactiae N14G was determined as a 1b serotype group that had genes for CPS L (688 bp), CPSG (621 bp), and CPS J (272 bp). The confirmation in tilapia of Aeromonas hydrophila and Streptococcus agalactiae by PCR and sequencing is important for enabling the detection of these organisms and also for the development of a combined vaccine to tackle co-infection.
      PubDate: 2022-01-03
      DOI: 10.4308/hjb.29.2.137-145
      Issue No: Vol. 29, No. 2 (2022)
  • Role of Fe2+-dependent Reaction in Biodecolorization of Methyl Orange by
           Brown-rot Fungus Fomitopsis pinicola

    • Authors: Adi Setyo Purnomo, Asranudin, Nela Rachmawati, Hamdan Dwi Rizqi, Refdinal Nawfa, Surya Rosa Putra
      Pages: 146 - 154
      Abstract: The involvement of Fenton reaction on biodegradation of methyl orange (MO) by brown-rot fungus Fomitopsis pinicola was investigated based on Fe2+-dependent reaction. The degradation of MO (final concentration 75 mg/L) was performed in mineral salt media with and without Fe2+ with incubation period at 0, 7, 14, 21, and 28 days. Degradation analysis was performed using UV-Vis Spectrophotometer and LC-TOF/MS. F. pinicola decolorized MO in a medium containing Fe2+ and a medium that lacked the mineral, at percentages of 89.47% and 80.08%, respectively. The optimum decolorization occurred after 28 days of incubation with the fungus on the presence of Fe2+, indicated that the presence of Fe2+ enhanced MO degradation with assumed to correlate with Fenton reaction. Two metabolites were detected through the LC-TOF/MS analysis, namely 4-(2-(4-(dimethyliminio)-2-hydroxycyclohexa-2,5-dien-1-ylidene) hydrazinyl) phenolate (m/z 258, RT: 1.28 min, compound 1) and 4-(2-(4-(dimethyliminio) cyclohexa-2,5-dien-1-ylidene) hydrazinyl) benzenesulfonate (m/z 391, RT: 2.70 min, compound 2). Compound 1 was a transformation product of hydroxylation and methylation, compound 2 was a product of dehydroxylation and desulfonation. This study indicated that the transformation of the metabolite structures was involved hydroxyl radical (OH.) and enzymatic mechanisms, which involved Fe2+-dependent reaction.
      PubDate: 2022-01-03
      DOI: 10.4308/hjb.29.2.146-154
      Issue No: Vol. 29, No. 2 (2022)
  • Genetic Diversity and Connectivity of Sea Urchin Tripneustes gratilla in
           Region Surrounding Cenderawasih Bay, Papua-Indonesia and Indo-Pacific

    • Authors: Abdul Hamid A. Toha, Ambariyanto Ambariyanto, Widodo Widodo, Luchman Hakim, Sutiman B. Sumitro, Agustina L. N. Aminin
      Pages: 155 - 163
      Abstract: Cenderawasih Bay has extremely high biodiversity with an enormous amount of endemic species due to geography isolation. Tripneustes gratilla is one of the species that is abundant in the bay. The species has ecological value that is suitable for bio-indicators of the environmental condition. Since the Bay relatively isolated area, then we examined the impact of the geographical conditions on genetic diversity and connectivity of Tripneustes gratilla among populations in the region surrounding Cenderawasih Bay based on the mitochondrial cytochrome c-oxidase-1 gene. Results of the study showed that genetic variation of the species within the population is high; despite the genetic variation among populations was low. The data suggested that the entire population of T. gratilla were closed connected, homogeneous, and shared polymorphic profile. Then we assumed the gene flow occurred for a long time among populations without geographical barriers. This information is a warrant to develop an effective strategy to maintain biodiversity in the Cenderawasih Bay.
      PubDate: 2022-01-03
      DOI: 10.4308/hjb.29.2.155-163
      Issue No: Vol. 29, No. 2 (2022)
  • Overproduction, Purification and Refolding of codon-optimized Hepatitis B
           Virus X Protein Subgenotype B3 in Escherichia coli BL21(DE3)

    • Authors: Anita Artarini, Armini Syamsidi, Anindyajati Anindyajati, Raymond R. Tjandrawinata, Debbie S. Retnoningrum
      Pages: 164 - 170
      Abstract: Hepatitis B virus (HBV) infects human and causes chronic liver infection, leading to liver cirrhosis and hepatocellular carcinoma. HBV X (Hbx) protein is known to interact with tumor suppressor protein p53 and block its translocation into the nucleus. This study outlines the overproduction of Hbx protein from HBV subgenotype B3 in Escherichia coli BL21(DE3), including its purification and refolding. The gene encoding Hbx was first codon-optimized and inserted into pET16b. The recombinant plasmid was then transformed into E. coli BL21(DE3) as an expression host. Optimization of Hbx expression was performed with variation of IPTG concentration and overproduction temperature. The results showed that Hbx protein was optimally induced by 0.075 mM IPTG and overproduction of Hbx at 17, 25, and 37°C exhibited no difference in protein level and location. The optimal refolding of Hbx was obtained using 0.1 M arginine prior to elution from Nickel column using 100 mM imidazole and 0.25 M arginine. Hbx migrates differently in SDS-PAGE reducing and non-reducing, while the melting curve pattern in TSA analysis changed after the refolding step. Essentially, this purified Hbx protein could potentially be used for interaction study with p53 and the inhibitor candidate of the protein.
      PubDate: 2022-01-17
      DOI: 10.4308/hjb.29.2.164-170
      Issue No: Vol. 29, No. 2 (2022)
  • The Structural Annotations of The Mir-122 Non-Coding RNA from The Tilapia
           Fish (Oreochromis niloticus)

    • Authors: Arli Aditya Parikesit, Imron Imron, Rizky Nurdiansyah, David Agustriawan
      Pages: 171 - 181
      Abstract: Tilapia (Oreochromis niloticus) is an important fisheries commodity. Scientific efforts have been done to increase its quality. One of them is staging a premium diet such as a fat-enriched diet. The transcriptomics approach is able to provide the signatures of the diet outcomes by observing the micro(mi)RNA signature in transcriptional regulation. Hence, it was found that the availability of mir-122 is essential in the regulation of a high-fat diet in tilapia. However, this transcriptomics signature is lacking structural annotations and the complete interaction annotations with its silencing(si)RNA. RNAcentral website was navigated for the latest annotation of mir-122 from tilapia and other species as a comparison. MEGA X was employed to comprehend the miRNA evolutionary repertoire. The RNA secondary structure prediction tools from the Vienna RNA package and the RNA tertiary structure prediction tools from simRNA and modeRNA are secured with default parameters. The HNADOCK tools were leveraged to observe the interaction between mir-122 and its siRNA. The post-processing was conducted with the Chimera visualization tool. The secondary and tertiary structure of the mir-122 and its siRNA could be elucidated, docked, and visualized. In this end, further effort to develop a comprehensive molecular breeding tool could be secured with the structural annotation information.
      PubDate: 2022-01-17
      DOI: 10.4308/hjb.29.2.171-181
      Issue No: Vol. 29, No. 2 (2022)
  • Leaf Litter Decomposition and Nutrient Release of Three Native Tree
           Species in a Drained Tropical Peatland in Riau, Indonesia

    • Authors: Ahmad Junaedi, Nina Mindawati, Avry Pribadi, Suryo Hardiwinoto
      Pages: 182 - 191
      Abstract: The decomposition and its nutrient release were the key ecological process that had a broad role in the forest ecosystem. This study aimed to investigate the leaf litter decomposition rate and its nutrient release of three native tree species of tropical peat swamp forest, namely Macaranga pruinosa, Macaranga gigantea, and Cratoxylum arborescens and one exotic species i.e Acacia crassicarpa. The decomposition and nutrient release were monitored in an experimental plot using litter bag technique. The initial litter quality of each litter and micro-environment properties were also observed. The result showed that the decomposition and its nutrient release were insignificantly different among native tree species and also between native species and Acacia crasssicarpa. The litter decomposition of all tree species was slow; with the range of k was 0.98-1.19 year-1. However, the P and K release from the decomposition of native species litter after four months of incubation were quickly, ranging 70-74% and 88-93%. We were suggested that the high of lignin content in the leaf litter (36-39%) was the main factor that made slow decomposition. These findings could be used as one of the tools in tree species selection for peat swamp forest rehabilitation.
      PubDate: 2022-01-17
      DOI: 10.4308/hjb.29.2.182-191
      Issue No: Vol. 29, No. 2 (2022)
  • Implementation of Dimer-based Screening System in Escherichia coli
           BL21(DE3) for Selection of Actinomycetes Compounds as Anti-HIV Candidate

    • Authors: Kenia Permata Sukma, Putri Cahya Destiani, Azzania Fibriani
      Pages: 192 - 203
      Abstract: Actinomycetes are reported to have inhibitory activity against several types of Human Immunodeficiency Virus proteases, enzyme with major role in the process of maturation of the virus thus it can infect new cells. Therefore, exploration of Indonesia’s actinomycetes species is expected to be a breakthrough for HIV treatment. In this study, selection of anti-HIV candidate compounds was conducted using a dimer-based screening system on recombinant Escherichia coli BL21(DE3). The construct includes the fusion of the AraC DNA binding domain + HIV-1 protease as the regulator and the green fluorescence protein as the reporter. Confirmation of the plasmid construct was carried out by PCR which showed size of ~1,076 bp. Sequencing analysis proved 100% similarity and identity between construct used in this study and one previously designed. SDS-PAGE showed the presence of band in the size of ~24 kDa equal to the size of the fusion protein. Compounds BLH 1-12 (2) EA, MAE 1-13 EA, BLH 1-1 EA, BLH 7-5 MetA, LC 98 (1) EA, exhibited consistent and significant protease-HIV inhibitory activity at certain concentrations. Thus, in this study, dimer-based screening system is considered to be able to detect actinomycetes as a new anti-HIV candidate for the protease inhibitor group.
      PubDate: 2022-01-18
      DOI: 10.4308/hjb.29.2.192-203
      Issue No: Vol. 29, No. 2 (2022)
  • Glycoengineering of Darbepoetin-α in CHO-DG44 Cells through
           Overexpression of α-2,3-sialyl-transferase and CMP-sialic Acid

    • Authors: Adi Santoso, Endah Puji Septisetyani, Ratna Dwi Ramadani, Yana Rubiyana, Pekik Wiji Prasetyaningrum, Popi Hadi Wisnuwardhani, Arizah Kusumawati, Neny Nuraini
      Pages: 204 - 213
      Abstract: Sialic acid plays a very important role in determining the circulation life span of glycoprotein in various organisms. Therefore, having a high content of sialic acid is needed by glycoprotein therapeutic agents to be able to function as desired. For example, Darbepoetin (DPO), the 5 N-linked erythropoietin showed higher bioavailability and efficacy compared to 3 N-linked erythropoietin. However, in the DPO production process, the molecular weight can vary and is highly dependent on the content of sialic acid and its production host. To improve the DPO sialic acid contents in our CHO-DG44 expressing DPO, we have engineered the cells through overexpression of α-2,3-sialyl-transferase (ST) and CMP-sialic acid transporter (CST). The DPO contained in the supernatant of the engineered cells was analyzed by Western blot and characterized by using PNGase-F or neuraminidase enzyme digestions. The results showed that, two clones, overexpressing ST or CST, were obtained. The clones showed higher molecular weight of DPO as compared to DPO expressed by the parental cells, yet retained the same protein backbone. The overexpression of these two genes does not affect cell growth. This suggests that may be these cells beneficial for therapeutic glycoproteins.
      PubDate: 2022-01-18
      DOI: 10.4308/hjb.29.2.204-213
      Issue No: Vol. 29, No. 2 (2022)
  • Antioxidant Activity of Endophytic Bacteria Derived from Hoya multiflora
           Blume Plant and Their Cellular Activities on Schizosaccharomyces pombe

    • Authors: Nabila Tsoerayya Gustia Pudjas, Nisa Rachmania Mubarik, Rika Indri Astuti, Lisdar I Sudirman
      Pages: 214 - 221
      Abstract: Endophytic bacteria isolated from plant tissues can produce the same secondary metabolites as their host plants. One of the metabolites that the bacteria can produce is antioxidants. This research aimed to analyze and measure the antioxidant activities of two endophytic bacteria, i.e. Bacillus siamensis HMB1 and Bacillus aryabhattai HMD4 cultures, derived from Hoya multiflora Blume plant, a tropical epiphytic plant species that grows in Indonesia, and to identify their cellular effects on Schizosaccharomyces pombe. The active compounds went through extraction process, and the antioxidant activities were measured, the extracts went under phytochemical analysis, and their phenol and flavonoid contents were measured. In vitro analysis was carried out using S. pombe. The results of this research indicated that both cultures had antioxidant activities, where HMB1 showed the highest IC50 value (51.18 mg/ml) among all. In vitro analysis indicated that HMD4 bacterial crude extract in 250 ppm concentration showed the highest resistance effect and significantly enhanced S. pombe growth. In addition, the results of the LC-MS analysis suggested that a total of 14 compounds potentially had antioxidant activity.
      PubDate: 2022-01-19
      DOI: 10.4308/hjb.29.2.214-221
      Issue No: Vol. 29, No. 2 (2022)
  • Intra- and Interspecies Wing Venation Variations of Apis cerana and Apis
           nigrocincta Species in Indonesia

    • Authors: Nisfia Rakhmatun Nisa, Berry Juliandi, Rika Raffiudin, Jauharlina Jauharlina, Mahardika Gama Pradana, Araz Meilin, Jasmi Jasmi, Yulia Pujiastuti, Puji Lestari, Fahri Fahri, Windra Priawandiputra, Tri Atmowidi
      Pages: 222 - 233
      Abstract: Apis cerana has a wide distribution in Asia, including Sundaland, and is currently found in Wallacea, while the sister species, A. nigrocincta, is native in Sulawesi. The wide geographic distribution and the island isolation led to form morphological differences in the bees. The morph and wing venations are known to have a high
      genetic inheritance. Therefore, this research aimed to (1) analyze the landmark variation of wing venations of A. cerana from Sundaland and Wallacea, and A. nigrocincta from Sulawesi, (2) determine the relationship between these two bee species. The research was conducted by digitizing 550 wing venations based on nineteen landmarks. Our study on intraspecies showed that A. cerana Sumatra revealed a high variation in bending energy. Overall, the deformation grid of A. cerana from Sundaland has higher displacement than those from Wallacea, meaning higher variations of the Sundaland A. cerana. We found geometric morphometric markers of landmarks 16 and 17 in intraspecies and interspecies bees. Thus, these landmarks known as a cubital index can be used for species identification. The differentiation of interspecies has been shown in the PCA. Apis nigrocincta was separated from the single group of the centroid A. cerana and was supported by the Neighbor-Joining tree.
      PubDate: 2022-01-26
      DOI: 10.4308/hjb.29.2.222-233
      Issue No: Vol. 29, No. 2 (2022)
  • Resistance of Nilaparvata lugens to Fenobucarb and Imidacloprid and
           Susceptibility to Neem Oil Insecticides

    • Authors: Danar Dono, Neneng S Widayani, Safri Ishmayana, Yusup Hidayat, Fitri Widiantini, Ceppy Nasahi
      Pages: 234 - 244
      Abstract: This study aims to monitor and determine the level of resistance of N. lugens to fenobucarb and imidacloprid, to analyse of the specific activities of asetylcholinesterase, Glutation S-Transferase, esterase, and to determine sensitivity of resistant N. lugens to the neem oil. Resistance tests were conducted on a field population from Cipunagara, Subang, West Java, Indonesia, and a standard population originating from Banyuwangi, East Java, that was obtained from the Indonesian Center for Rice Research. Synthetic and neem Insecticides were applied using the leaf-stem dipping method. The resistance ratio obtained by comparing the LC50 value of the field population with the standard. The test insects were considered to be resistant to synthetic insecticide if they had a resistance ratio (RR) of more than 4, indicated resistance if RR ≥1, and susceptible to neem if has RR <1. The results of the research showed that the N. lugens population of Cipunagara indicated resistance to fenobucarb (RR = 2.43), while it was clearly resistance to imidacloprid (RR = 13.95). Specific activities of the acetylcholinesterase and Glutathione S-Transferase of the N. lugens Cipunagara populations were higher than the standard, while the esterase specific activity was lower. These two enzymes are responsible for the resistance of N. lugens to fenobucarb and imidacloprid. Nevertheless, the N. lugens population of Cipunagara was susceptible to neem (RR <1) that never been reported before. Therefore, it is important to know the change in the sensitivity of the target and the metabolic pathway of the insecticide used to manage insect resistance.
      PubDate: 2022-01-26
      DOI: 10.4308/hjb.29.2.234-244
      Issue No: Vol. 29, No. 2 (2022)
  • Reef Fish in the Mudflats of Kaledupa Island in Wakatobi National Park,

    • Authors: Ernik Yuliana, Adi Winata, Hasan Eldin Adimu, Yuni Tri Hewindati, Wibowo A. Djatmiko
      Pages: 245 - 254
      Abstract: Although frequently described as low-fertility or low-productivity habitat, coastal mudflats serve as important feeding grounds for fish. Many fish species from adjacent coral reefs, seagrass beds, or mangroves foraging periodically in mudflats. Because of this foraging behaviour, some local fishermen are known to utilize the mudflats to catch fish. However, the impact of this catching activities to the ecosystem has not been fully discovered. An examination of the fish community structure and levels of environmental stress had carried out in the mudflat ecosystem of the coast of Kaledupa Island in Wakatobi National Park (WNP), Indonesia. Two mudflat study sites were selected from the shore of Balasuna and Tampara villages located between mangroves and coral reefs. Data were sampled from the fish catch of local fishermen using fish fences (sero) installed in each mudflat area. Fish community structure was analyzed using diversity index and index of relative importance (IRI). ABC curves and species exploitation rate were used to assess the local environmental pressure. A total of 74 fish species were recorded from the mudflats of Kaledupa, which was found to be dominated by reef-associated fish species, comprising 63 species and accounting for 85% of the total catch. Additionally, although both sites had relatively high reef fish diversity, the obtained Clarke’s W-statistic values were approximately 0, indicating that the local fish communities presented moderate levels of disturbance. Three out of five fish species with the highest IRI values were found to be over-exploited, namely Siganus canaliculatus, Lethrinus ornatus, and Lethrinus variegatus.
      PubDate: 2022-01-26
      DOI: 10.4308/hjb.29.2.245-254
      Issue No: Vol. 29, No. 2 (2022)
  • The Fishing Grounds and the Exploitation Status of Kawakawa (Euthynnus
           affinis) in Java Sea, Indonesia

    • Authors: Siti Mardlijah, Andina Ramadhani Putri Pane, Moh Fauzi, Helman Nur Yusuf, Heri Widiyastuti, Herlisman Herlisman, Achmad Zamroni, Tegoeh Noegroho, Hufiadi Hufiadi, Karsono Wagiyo
      Pages: 255 - 265
      Abstract: Kawakawa (Euthynnus affinis) is one of the highly favored mackerel tunas caught in Java Sea. The increase in demand due to local consumption and export eventually increases fishing that is proportional to the exploitation status. To maintain the fish resource, a scientific study should be carried out on the fishing grounds and the exploitation status of kawakawa. This study was carried out for 3 years, i.e. in March to October 2017, June to December 2018, and February to November 2019 at Pekalongan National Fishing Port in Pekalongan, Central Java, with 9,511 fish as the study objects. The data analysis was carried out by observing the fishing grounds using GPS and through interview. In addition, the exploitation status was analyzed using FISAT II application and ELEFAN program. The fishing grounds were mostly coastal waters with 15–55 cm sized kawakawa. The size of the first caught (Lc) of the kawakawa in this area ranged from 26.9 cm to 38.4 cm. The fish recruitment occurred all year long, peaking in Apr at 15.97% and Juli 13.62%.The fish natural mortality (M) was smaller than its fishing mortality (F), while its exploitation status was 0.66 (overfished), meaning that the current fishing efforts should be reduced by 32%. The other management efforts that can be carried out are among others conservation, keeping the fishing grounds outside the spawning grounds, and controlling the mesh size.
      PubDate: 2022-01-26
      DOI: 10.4308/hjb.29.2.255-265
      Issue No: Vol. 29, No. 2 (2022)
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