Molecular Cloning and Characterization of a Ferritin Heavy Chain Subunit of the Cricket Teleogryllus emma

Mi Ri Sohn,Nam Jung Kim,Hung Dae Sohn,Byung Rae Jin 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05

We describe here the cloning and characterization of a cDNA encoding the ferritin heavy chain homologue (TeFerHCH) from the cricket Teleogryllus emma. The TeFerHCH gene spans 1,009 bp and consisted of four introns and five exons coding for 217 amino acids residues. The TeFerHCH subunit contained the conserved motifs for the ferroxidase center typical of vertebrate ferritin heavy chains and the iron-responsive element (IRE) sequence with a predicted stem-loop structure was present in the 5'-untranslated region (UTR) of TeFerHCH mRNA. TeFerHCH was grouped with the S type (HCH) in a phylogenetic tree. The TeFerHCH cDNA was expressed as approximately 27 kDa polypeptide in baculovirus-infected insect Sf9 cells. Northern blot analysis revealed that TeFerHCH exhibited ubiquitous expression and was upregulated by wounding and iron overload in the fatbody, suggesting a functional role for TeFerHCH in iron metabolism.

Production of Classical Swine Fever Virus Envelope Glycoprotein E2 as a Granule-Form Antigen in Baculovirus-Infected Silkworm Larvae

Mi Ri Sohn,Kwang Sik Lee,Bo Yeon Kim,Young Moo Choo,Soo Dong Woo,Sung Sik Yoo,Yeon Ho Je,Jae Young Choi,Jong Yul Roh,Hyun Na Koo,Young In Kim,Byung Rae Jin 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10

Classical swine fever virus (CSFV) envelope glycoprotein E2 is the main target for inducing neutralizing antibodies and protective immunity in swine. Here, we report a novel strategy forthe large-scale production of a CSFV E2 subunit vaccine that demonstrates a high immunogenic capability in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (CSFV E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed an approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Mice that were immunized with the granule form of recombinant polyhedra or the soluble form of the fusion protein elicited CSFV E2 antibodies, which indicated that the recombinant polyhedra carrying CSFV E2ΔC were immunogenic. The virus neutralization test showed that the serum from mice that were treated with recombinant polyhedra or the soluble form of the fusion protein contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen and that the recombinant polyhedra containing CSFV E2ΔC as a granule antigen can be used as a potential subunit vaccine against CSFV.

Production and Immunogenicity of Classical Swine Fever Virus Envelope Glycoprotein E2 as Recombinant Polyhedra in Baculovirus-Infected Silkworm Larvae

Mi Ri Sohn,Kwang Sik Lee,Byung Rae Jin 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.05

The silkworm-baculovirus expression system has distinct advantages, such as a high yield and safe usage in vertebrates. Here, we report a novel strategy for the large-scale production of a classical swine fever virus (CSFV) envelope glycoprotein E2 in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed native polyhedrin and approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra contained both the fusion protein and native polyhedrin were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg per ml of hemolymph and 0.53 mg per larva at 6 days post-infection. Six-week-old female BALB/c mice that were immunized with the E2ΔC protein purified from solubilized recombinant polyhedraelicited CSFV E2 antibodies, which indicated that the CSFV E2ΔC protein from recombinant polyhedra was immunogenic. The virus neutralization test showed that the serum from mice that were treated with E2ΔC protein from recombinant polyhedra contained significant levels of virus neutralization activity. These results demonstrate that the present strategy can be used for the large-scale production of CSFV E2 antigen.

Molecular characterization of a venom serine protease from the bumblebee Bombus terrestris

Yuling Qiu,Young Moo Choo,Mi Ri Sohn,Hyung Joo Yoon,Hung Dae Sohn,Byung Rae Jin 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05

Bee venom contains a variety of peptides and enzymes, including serine proteases. Here we describe the molecular cloning and characterization of a serine protease (Bt-VSP) isolated from the venom of the bumblebee Bombus terrestris. The Bt-VSP gene consists of six exons encoding a 358-amino acid protein. The form of Bt-VSP detected in bee venom was the 34-kDa mature protein, which is created by cleavage of the catalytic domain of Bt-proVSP between Arg111 and Val112. Bt-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles for Bt-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. The finding that Bt-VSP acts as a fibrin(ogen)olytic enzyme is similar to a previous finding that Bi-VSP, a venom serine protease of B. ignitus, exhibits fibrin(ogen)olytic activity. We also compared major venom components in honeybee and bumblebee, and found that bumblebee venom contains a larger amount of serine protease. Furthermore, unlike bumblebee venom, which exhibits fibrin(ogen)olytic activity owing to the presence of a serine protease, it is likely that honeybee venom lacks fibrin(ogen)olytic activity.

Molecular cloning and characterization of 1-Cys and 2-Cys peroxiredoxins from the bumblebee Bombus ignitus

Kwang Sik Lee,Zhigang Hu,Young Moo Choo,Mi Ri Sohn,Hyung Joo Yoon,Sang Mong Lee,Hung Dae Sohn,Byung Rae Jin 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10

We cloned and characterized two peroxiredoxins (Prxs), BiPrx1 (a 1-Cys Prx) and BiTPx1 (a 2-Cys Prx) from the bumblebee Bombus ignitus. The BiPrx1 gene consists of 5 exons, encoding 220 amino acid residues with one conserved cysteine residue. The BiTPx1 gene consists of three exons, encoding 195 amino acid residues with 2 conserved cysteine residues. Recombinant BiPrx1 (27 kDa) and BiTPx1 (25 kDa), expressed in baculovirus-infected insect Sf9 cells, reduced H2O2 in the presence of electrons donated by dithiothreitol. Unlike BiTPx1, however, BiPrx1 did not show reduction activity when thioredoxin was used as the electron donor. Both BiPrx1 and BiTPx1 protected super-coiled DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Tissue distribution analyses showed the presence of BiPrx1 and BiTPx1 in the fat body, midgut, muscle and epidermis, but not in the hemolymph, suggesting that BiPrx1 and BiTPx1 are not secretable. When H2O2 was injected into B. ignitus bees, BiPrx1 and BiTPx1 transcripts were acutely up-regulated in the fat body tissues. We also demonstrated regulation of BiPrx1 and BiTPx1 expression via reduction of transcript levels in the fat body with RNA interference (RNAi). Under H2O2 overload, the RNAi-induced BiPrx1 knock-down B. ignitus worker bees showed up-regulated expression of BiTPx1. Reciprocally, BiTPx1 RNAi knockdowns showed up-regulated BiPrx1 expression in the fat body. These results indicate that loss of expression of BiPrx1 or BiTPx1 is compensated by up-regulation of expression of the other peroxidase in response to H2O2 overload.

Molecular Cloning and Oxidative Stress Response of a Sigma-Class Glutathione S-Transferase of the Bumblebee Bombus ignitus

Bo Yeon Kim,Kwang Sik Lee,Hu Wan,Mi Ri Sohn,Yuling Qiu,Hyung Joo Yoon,Byung Rae Jin 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainlyinvolved in the xenobiotic metabolism and protection against oxidative damage. Most studies of GSTs in insects have been focused on their role in detoxifying exogenous compounds in particular insecticides. Here, we show the expression profiles of GSTs of the bumblebee Bombus ignitus in response to oxidative stress. We identified a sigma-class GST from B. ignitus (BiGSTS). The BiGSTSgene consists of 4 exons that encode 201 amino acids. Comparative analysis indicates that the predicted amino acid sequence of BiGSTS shares a high identity with the sigma-class GSTs of hymenopteran insects such as Apis mellifera (70% protein sequence identity) and Solenopsis invicta (59% protein sequence identity). Tissue distribution analyses showed the presence of BiGSTS in all tissues examined, including the fat body, midgut, muscle and epidermis. The oxidative stress responses analyzed by quantitative real-time PCR showed that under H2O2 overload, BiGSTS and BiGSTD (identified in our previous study) were upregulated in all tissues examined, including the fat body and midgut of B. ignitus worker bees. Under uniform conditions of H2O2 overload, the expression profile of GSTs and other antioxidant enzyme genes, such as phospholipid-hydroperoxide glutathione peroxidase (Bi-PHGPx) and peroxiredoxins (BiPrx1 and BiTPx1), showed that other antioxidant enzyme genes are acutely induced at 3 h after H2O2 exposure, whereas BiGSTS and BiGSTD are highly induced at 9 h after H2O2 exposure in the fat body of B. ignitus worker bees. These findings indicate that GSTs and other antioxidant enzyme genes in B. ignitusare differentially expressed in response to oxidative stress. Taken together, our findings indicate that BiGSTS and BiGSTD are oxidative stress-inducible antioxidant enzymes that may play a role in oxidative stress response.

Biological Effects of Different Thin Layer Hydroxyapatite Coatings on Anodized Titanium

Sohn, Sung-Hwa,Jun, Hye-Kyoung,Kim, Chang-Su,Kim, Ki-Nam,Ryu, Yeon-Mi,Lee, Seung-Ho,Kim, Yu-Ri,Seo, Sang-Hui,Kim, Hye-Won,Shin, Sang-Wan,Ryu, Jae-Jun,Kim, Meyoung-Kon The Korean Society of Toxicogenomics and Toxicopro 2005 Molecular & cellular toxicology Vol.1 No.4

Several features of the implant surface, such as roughness, topography, and composition play a relevant role in implant integration with bone. This study was conducted in order to determine the effects of various thin layer hydroxyapatite (HA) coatings on anodized Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on A (100 nm HA coating on anodized surface), B (500-700 nm HA coating on anodized surface), C ($1{\mu}m$ HA coating on anodized surface), and control (non HA coating on anodized surface) Ti. The morphology of these cells was assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1,152 elements). The appearances of the surfaces observed by SEM were different on each of the four dental substrate types. MG63 cells cultured on A, C and control exhibited cell-matrix interactions. It was B surface showing cell-cell interaction. In the expression of several genes were up-, and down-regulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface morphology of the dental materials used.

Small-Dose India Ink Tattooing for Preoperative Localization of Colorectal Tumor

Hwang, Mi Ri,Sohn, Dae Kyung,Park, Ji Won,Kim, Byung Chang,Hong, Chang Won,Han, Kyung Su,Chang, Hee Jin,Oh, Jae Hwan Mary Ann Liebert 2010 Journal of Laparoendoscopic & Advanced Surgical Te Vol.20 No.9

RNA-RNA Interactions between RNA Elements at the 5' end and at the Upstream of sgRNA of RNA Genome are Required for Potato virus X RNA Replication

Park, Mi-Ri,Park, Sang-Ho,Cho, Sang-Yun,Hemenway, Cynthia L.,Choi, Hong-Soo,Sohn, Seong-Han,Kim, Kook-Hyung The Korean Society of Plant Pathology 2008 Plant Pathology Journal Vol.24 No.3

RNA-RNA interactions and the dynamic RNA conformations are important regulators in virus replication in several RNA virus systems and may also involved in the regulation of many important virus life cycle phases, including translation, replication, assembly, and switches in these important stages. The 5' non-translated region of Potato virus X(PVX) contains multiple cis-acting elements that facilitate various viral processes. It has previously been proposed that RNA-RNA interactions between various RNA elements present in PVX RNA genome are required for PVX RNA accumulation(Hu et al., 2007; Kim and Hemenway, 1999). This model was based on the potential base-pairing between conserved sequence elements at the upstream of subgenomic RNAs(sgRNAs) and at the 5' and 3' end of RNA genome. We now provide more evidence that RNA-RNA base-pairing between elements present at the 5' end and upstream of each sgRNA is required for efficient replication of genomic and subgenomic plus-strand RNA accumulation. Site-directed mutations introduced at the 5' end of plus-strand RNA replication defective mutant(${\Delta}12$) increasing base-pairing possibility with conserved sequence elements located upstream of each sgRNAs restored genomic and subgenomic plus-strand RNA accumulation and caused symptom development in inoculated Nicotiana benthamiana plants. Serial passage of a deletion mutant(${\Delta}8$) caused more severe symptoms and restored wild type sequences and thus retained possible RNA-RNA base-pairing. Altogether, these results indicate that the RNA element located at the 5' end of PVX genome involved in RNA-RNA interactions and play a key role in high-level accumulation of plus-strand RNA in vivo.

대사증후군 환자의 영양소 섭취상태 및 식사의 질과 염증지표 농도의 상관성

김미성(Mi Sung Kim),김주영(Ju Young Kim),김소혜(So Hye Kim),배우경(Woo Kyung Bae),손정민(Cheong Min Sohn),이예송(Ye Song Lee),나우리(Woo Ri Na) 대한지역사회영양학회 2011 대한지역사회영양학회지 Vol.16 No.1

Elevated serum concentration of inflammation markers is known as an independent risk factor of metabolic syndrome (MS) and dietary intake is an important factor to control MS. The purpose of this study was to investigated the hypothesis that inflammatory indices are associated with dietary intake and diet quality index-international (DQI-I) in subjects with MS. A cross-sectional study was conducted on 156 men and 73 postmenopausal women with MS, defined by three or more risk factors of the modified Adult Treatment Panel III criteria. Serum levels of high sensitive C-reactive protein (hs-CRP), adiponectin were examined and nutrients intake and DQI-I were assessed using a semiquantitative food frequency questionnaire. The total DQI-I score was significantly higher in female subjects (65.87 ± 9.86) than in male subjects (62.60 ± 8.95). There was a positive association between hs-CRP and polyunsaturated fatty acid intake (p < 0.05) and a negative association between adiponectin and lipid (p < 0.05), total sugar (p < 0.01), and total fatty acids (p < 0.05). When the subjects were divided into 5 groups by quintile according to serum adiponectin and hs-CRP level, there was no association between DQI-I score and hs-CRP levels. Moderation score of DQI-I was significantly higher in highest quintile group than the lower quintile groups. Therefore, our results provide some evidence that dietary intake and diet quality are associated with inflammation markers and dietary modification might be a predictor to decrease risk for metabolic syndrome complications. However further research is needed to develop the dietary quality index reflecting the inflammatory change by considering the dietary habit and pattern of Koreans. (Korean J Community Nutr 16(1) : 51~61, 2011)