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- 검색키워드
- 저자 : Zoysa, (검색결과 115 건)
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De Zoysa, M.,Nikapitiya, C.,Kim, Y.,Oh, C.,Kang, D.H.,Whang, I.,Kim, S.J.,Lee, J.S.,Choi, C.Y.,Lee, J. Academic Press 2010 Fish & shellfish immunology Vol.29 No.2
Here, we report the identification and characterization of allograft inflammatory factor-1 (AIF-1) from disk abalone Haliotis discus discus that was denoted as AbAIF-1. The full-length cDNA of AbAIF-1 consists of a coding region (453 bp) for 151 amino acids with a 17 kDa molecular mass. Analysis of AbAIF-1 sequence showed that it shares characteristic two EF hand Ca<SUP>+2</SUP>binding motifs. Results from phylogenetic analysis further confirm that AbAIF-1 is a member of the AIF-1 family similar to invertebrate and vertebrate counterparts suggesting it has high evolutional conservation. Tissue-specific expression and transcriptional regulation of AbAIF-1 were analyzed after bacteria (Vibrio alginolyticus, Vibrio parahemolyticus and Lysteria monocytogenes), viral hemorrhagic septicemia virus (VHSV) immune challenge and during tissue injury by quantitative real-time PCR. It is shown that the expression of AbAIF-1 mRNA was expressed ubiquitously in all selected tissues in constitutive manner showing the highest level in hemocytes. Upon bacteria and VHSV challenge, AbAIF-1 showed the significant up-regulation in hemocytes than gills. After the tissue injury in shell and mantle, AbAIF-1 and antioxidant selenium-dependant glutathione peroxidase (SeGPx) transcripts were significantly upregulated in abalone hemocytes. Taken together, these findings suggest that AIF-1 could response against the pathogenic challenge or tissue injury in abalone like mollusks. Also, AbAIF-1 may involve in wound healing and shell repair after the tissue injury of abalone.
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De Zoysa, M.,Nikapitiya, C.,Oh, C.,Whang, I.,Lee, J.S.,Jung, S.J.,Choi, C.Y.,Lee, J. Academic Press 2010 FISH AND SHELLFISH IMMUNOLOGY Vol.28 No.5-6
The lipopolysaccharide-induced TNF-α factor (LITAF) and Rel family nuclear factor kappaB (Rel/NF-kB) are two important transcription factors which play major roles in the regulating inflammatory cytokine, apoptosis and immune related genes. Here, we report the discovery of disk abalone LITAF (AbLITAF) and Rel/NF-kB (AbRel/NF-kB) homologues and their immune responses. Full-length cDNA of AbLITAF consists of 441 bp open reading frame (ORF) that translates into putative peptide of 147 aa. Analysis of AbLITAF sequence showed it has characteristic LITAF (Zn<SUP>+2</SUP> binding domain with two CXXC motifs. Phylogenetic analysis results further revealed that AbLITAF is a member of LITAF family. AbRel/NF-kB is 584 aa protein that contains several characteristic motifs including Rel homology domain (RHD), Rel protein signature, DNA binding motif, nuclear localization signal (NLS) and transcription factor immunoglobulin - like fold (TIG) similar to their invertebrate and vertebrate counterparts. Tissue specific analysis results showed that both AbLITAF and AbRel/NF-kB mRNA was expressed ubiquitously in all selected tissues in constitutive manner. However, constitutive expression of AbLITAF was higher than AbRel/NF-kB in all tissues except mantle. Upon immune challenge by bacteria (Vibrio alginolyticus, Vibrio parahemolyticus and Lysteria monocytogenes) and viral hemoragic septicemia virus (VHSV), AbLITAF showed the significant up-regulation in gills while AbRel/NF-kB transcription was not change significantly. Based on transcriptional response against immune challenge, we could suggest that regulation of TNF-α expression may have occurred mainly by LITAF activation rather than NF-kB in disk abalone. The cumulative data from other molluscs and our data with reference to TNF-α, LITAF and Rel/NF-kB from disk abalone provide strong evidence that LITAF and NF-kB are independent pathways likely to occur throughout the Phylum mollusca.
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De Zoysa, M.,Whang, I.,Lee, Y.,Lee, S.,Lee, J.S.,Lee, J. Academic Press 2010 FISH AND SHELLFISH IMMUNOLOGY Vol.28 No.2
Gene-encoded antimicrobial peptides (AMPs) serve a major role in host defense systems against pathogens. In this study, cDNA of a new mollusk defensin was identified from a normalized cDNA library constructed from whole tissues of disk abalone. Abalone defensin peptide (pro-defensin) has a 198-bp coding sequence comprised of a putative 66 amino acids with a mature defensin consisting of 48 amino acid residues. The presence of an invertebrate defensin family domain, an arrangement of six cysteine residues and their disulfide linkage in C<SUB>1</SUB>-C<SUB>4</SUB>, C<SUB>2</SUB>-C<SUB>5</SUB> and C<SUB>3</SUB>-C<SUB>6</SUB> form, an alpha helix in three-dimensional structure and a phylogenetic relationship suggests that abalone defensin could be a new member of the invertebrate defensin family, and related to arthropod defensins. In non-stimulated abalone, defensin transcripts were constitutively expressed in all examined tissues including hemocytes, gills, mantle, muscle, digestive tract and hepatopancreas. It was observed that abalone defensin transcripts were significantly induced in hemocytes, gills and digestive tract at different time intervals after infection by pathogenic bacteria mixture containing Vibrio alginolyticus, Vibrio parahemolyticus and Lysteria monocytogenes. Our overall results suggest that disk abalone defensin could be involved in the immune response reactions as a host defense against pathogenic bacteria.
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De Zoysa, M.,Lee, J. Academic Press 2009 FISH AND SHELLFISH IMMUNOLOGY Vol.26 No.3
Suppressor of cytokine signaling (SOCS) proteins are inhibitors of cytokine signaling pathways and key physiological regulators of both innate and adaptive immune systems in vertebrates. In present study, we provided the initial characterization of SOCS-2 homologue from the mollusk abalone Haliotis discus discus as a member of invertebrate SOCS-2. The disk abalone SOCS-2 cDNA (AbSOCS-2) contains 1700-bp full length with 771-bp coding sequence, which codes 257 amino acids protein. Based on the sequence analysis results, AbSOCS-2 showed characteristic SH2 domain and SOCS box similar to vertebrate SOCS counterparts. Additionally, 3' UTR of the AbSOCS-2 contained two RNA instability motifs (ATTTA). Quantitative real-time PCR expression results showed that AbSOCS-2 was constitutively expressed in all examined tissues of healthy abalone showing tissue specific variation. AbSOCS-2 mRNA expression was induced significantly (p<0.05) by bacteria mixture containing Vibrio alginolyticus, Vibrio parahemolyticus, and Listeria monocytogenes in abalone gill at 3h post-induction (p.i.). Furthermore, AbSOC-2 mRNA was significantly (p<0.05) induced by viral haemorrhagic septicemia virus (VHSV) and lipopolysaccharide (LPS) treatments in abalone gill. Therefore, discovery and expression analysis of abalone SOCS-2 gene would provide evolutional relationship of SOCS family members and immune defense responses against bacteria, virus and LPS like immune modulators.
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Collective Forest Management System in Japan: a Case Study in Osawa Property Ward Forest
De Zoysa, Mangala Premakumara,Inoue, Makoto,Yamashita, Utako,Hironori, Okuda Institute of Forest Science 2013 Journal of Forest Science Vol.29 No.1
Iriai an Indigenous forest management system in Japan from the viewpoint of "common pool resources" was a success resilient institution and resulted with sustainable production system and environmental conservation. This study was conducted in Osawa of the Nagano prefecture through group discussions, field observations and an in-depth field survey. Osawa Property Ward Forest is managed under the concept very much similarly to traditional "Iriai". This study firstly examined the changes of collective forest management system in terms of awareness and interest in forest management; forest management activities; role of forest; and collection of forest products. Then it analyzed the current threats for collective forest management have been identified as: land abandonment due to loss of benefits and lack of active community participation; deterioration of forest environment particularly the micro-climate and aesthetic values; conflict with local government authorities restraining the use of money in property ward forest and conflict with outsiders on damping of the garbage. Community cantered forestry management rules; livelihood contribution; protection of environment; local initiatives for protection and economic activities are the prevailing opportunities for collective forest management. The main requirements for revitalization of collective forest management are explained as local reciprocity; imposition of community based forest rules; encouraging local innovations; and building partnerships with stakeholders. Collective forest management system addresses the limitations of conventional forestry models, which had invalidated traditional 'iriai' institutions, and key to restoring sustainable use of forest and environmental resources. Cross-institutional collaborations together with responsibilities of local communities would ensure the revitalization of forest resources.
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De Zoysa, M.,Pushpamali, W.A.,Oh, C.,Whang, I.,Kim, S.J.,Lee, J. Academic Press 2008 FISH AND SHELLFISH IMMUNOLOGY Vol.25 No.4
Selenium dependent glutathione peroxidase (Se-GPx) belongs to the family of selenoprotein, which acts mainly as an antioxidant in the cellular defence system. We have identified Se-GPx full length cDNA from disk abalone (Haliotis discus discus) designated as AbSe-GPx. It has a characteristic codon at <SUP>223</SUP>TGA<SUP>225</SUP> that corresponds to selenocysteine (Sec) amino acid as U<SUB>75</SUB>. The full length cDNA consists of 675bp, an open reading frame encoding 225 amino acids. Sequence characterization revealed that AbSe-GPx contains a characteristic GPx signature motif 2 (<SUP>97</SUP>LGFPCNQF<SUP>104</SUP>), an active site motif (<SUP>183</SUP>WNFEKF<SUP>188</SUP>) and essential residues for the enzymatic function. Additionally, the eukaryotic selenocysteine insertion sequence (SECIS) is conserved in the 3' UTR. The AbSe-GPx amino acid sequence exhibited the highest level of identity (46%) with insect (Ixodes scapularis) GPx, and shares 41% with bivalve (Unio tumidus) Se-GPx. The RT-PCR analysis revealed that AbSe-GPx mRNA was expressed constitutively in gill, mantle, gonad, abductor muscle, digestive tract, and hemocytes in a tissue specific manner. AbSe-GPx mRNA expression was significantly up-regulated in gill and digestive tract tissues after H<SUB>2</SUB>O<SUB>2</SUB> injection and Vibrio alginolyticus infection. However, AbSe-GPx expression was not up-regulated after Aroclor 1254 injection. These results indicate that AbSe-GPx mRNA is expressed at a basal level in abalone tissues, which can be up-regulated transcriptionally by H<SUB>2</SUB>O<SUB>2</SUB> oxidative stress and Vibrio alginolyticus infection. Therefore, AbSe-GPx may be involved in a protective role against H<SUB>2</SUB>O<SUB>2</SUB> oxidative stress and immune defence against bacterial infection.
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De Zoysa, M.,Lee, J. Academic Press 2007 Fish & Shellfish Immunology Vol.23 No.3
Ferritin plays a key role in cellular iron metabolism, which includes iron storage and detoxification. From disk abalone, Haliotis discus discus, the cDNA that encodes the two ferritin subunits abalone ferritin subunit 1 (Abf1) and abalone ferritin subunit 2 (Abf2) were cloned. The complete cDNA coding sequences for Abf1 and Abf2 contained 621 and 549bp, encoding for 207 and 183 amino acid residues, respectively. The H. discus discus Abf2 subunit contained a highly conserved motif for the ferroxidase center, which consists of seven residues of a typical vertebrate heavy-chain ferritin with a typical stem-loop structure. Abf2 mRNA contains a 27bp iron-responsive element (IRE) in the 5'UTR position. This IRE exhibited 96% similarity with pearl and Pacific oyster and 67% similarity with human H type IREs. However, the Abf1 subunit had neither ferroxidase center residues nor the IRE motif sequence; instead, it contained iron-binding region signature 2 (IBRS) residues. Recombinant Abf1 and Abf2 proteins were purified and the respective sizes were about 24 and 21kDa. Abf1 and Abf2 exhibited iron-chelating activity 44.2% and 22.0%, respectively, at protein concentration of 6μg/ml. Analysis of tissue-specific expression by RT-PCR revealed that Abf1 and Abf2 ferritin mRNAs were expressed in various abalone tissues, such as gill, mantle, gonad, foot and digestive tract in a wide distribution profile, but Abf2 expression was more prominent than Abf1.
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De Zoysa, M.,Nikapitiya, C.,Moon, D.O.,Whang, I.,Kim, G.Y.,Lee, J. Academic Press 2009 FISH AND SHELLFISH IMMUNOLOGY Vol.27 No.3
Fas ligand is a member of the TNF superfamily that plays an important role by inducing apoptosis and homeostasis of immune responses. The gene encoding Fas ligand was isolated from a disk abalone (Haliotis discus discus) cDNA library, denoted as the AbFas ligand. It contains an 1832bp transcript with a 945bp open reading frame, encoding 315 amino acids. The AbFas ligand showed characteristic transmembrane and TNF family signature domains. The deduced amino acid comparison showed that the AbFas ligand exhibits 22.0, 16.1 and 14.5% identities to human Fas ligand, TNF-α, and lymphotoxin (LT-α), respectively. Phylogenetic analysis indicates that the AbFas ligand belongs to the invertebrate TNF family and it is closely related to vertebrate Fas ligand counterparts. Quantitative real-time PCR analysis results showed that the AbFas ligand transcripts were constitutively expressed in abalone hemocytes, gills, mantle, muscle, digestive tract and digestive gland in a tissue-specific manner. By immune stimulation, AbFas ligand mRNA was significantly (p<0.05) up-regulated after infection with a mixture of bacteria (Vibrio alginolyticus, Vibrio parahemolyticus, and Listeria monocytogenes), viral haemorrhagic septicaemia virus (VHSV), and lipopolysaccharide (LPS) in abalone gills. The recombinant AbFas ligand was over-expressed in Escherichia coli (E. coli) and purified using a pMAL protein fusion system. This recombinant AbFas ligand showed its biological activity by inducing both superoxide anion (O<SUB>2</SUB><SUP>-</SUP>) and H<SUB>2</SUB>O<SUB>2</SUB> in human THP-1 cells in concentration-dependant manner. Correlating the AbFas ligand transcriptional up-regulation against bacteria, virus and LPS with the biological activity of its recombinant protein, we could suggest that the abalone Fas ligand may control microbial infection by inducing O<SUB>2</SUB><SUP>-</SUP>, H<SUB>2</SUB>O<SUB>2</SUB> and other ROS.
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