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Expression of Antibacterial Protein, Nuecin, Using Baculorivus Expression Vector System in Bm5 Insect Cell and Bombyx mori (누에 배양세포(Bm5) 및 생체에서 베큘로바이러스 발현계를 이용한 누에신 단백질 발현 특성)

  • 윤은영;구태원;황재삼;김상현;강석우;김근영;진병래
    • Journal of Sericultural and Entomological Science
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    • v.44 no.2
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    • pp.69-73
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    • 2002
  • For the practical use of nuecin protein, we tried to overexpress nuecin using Bm5 insect cell and Bombyx mori. We inserted nuecin cDNA into pBm10po1-Xa vector derived from B. mori nuclear polyhedrosis virus (BmNPV), and expressed in Bm5 cells and B. mori respectively. SDS-PAGE and Northern blot analysis showed an expressed of the protein when baculovirus expression vector system (BEVS) was used. The amount of intracellular protein is abundant, but the amount of extracellular protein is poor. The results suggest that the biologically active nuecin protein produced by using BEVS is poor because incresed level of misfolded nuecin by the strong promoter, polyhedrin and p 10 of BEVS, decrease the level of free chaperons and foldases by binding with them.

Bacterial Toxin-antitoxin Systems and Their Biotechnological Applications (박테리아의 toxin-antitoxin system과 생명공학기술 응용)

  • Kim, Yoonji;Hwang, Jihwan
    • Journal of Life Science
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    • v.26 no.2
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    • pp.265-274
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    • 2016
  • Toxin-antitoxin (TA) systems are ubiquitous genetic modules that are evolutionally conserved in bacteria and archaea. TA systems composed of an intracellular toxin and its antidote (antitoxin) are currently classified into five types. Commonly, activation of toxins under stress conditions inhibits diverse cellular processes and consequently induces cell death or reversible growth inhibition. These effects of toxins play various physiological roles in such as regulation of gene expression, growth control (stress response), programmed cell arrest, persister cells, programmed cell death, phage protection, stabilization of mobile genetic elements or postsegregational killing of plasmid-free cells. Accordingly, bacterial TA systems are commonly considered as stress-responsive genetic modules. However, molecule screening for activation of toxin in TA system is available as development of antimicrobial agents. In addition, cytotoxic effect induced by toxin is used as effective cloning method with antitoxic effect of antitoxin; consequently cells containing cloning vector inserted a target gene can survive and false-positive transformants are removed. Also, TA system is applicable to efficient single protein production in biotechnology industry because toxins that are site-specific ribonuclease inhibit protein synthesis except for target protein. Furthermore, some TA systems that induce apoptosis in eukaryotic cells such as cancer cells or virus-infected cells would have a wide range of applications in eukaryotes, and it will lead to new ways of treating human disease. In this review, we summarize the current knowledge on bacterial TA systems and their applications.

The role of Fatty acid binding protein 5 (Fabp5) in fatty acid partitioning in the liver (간에서 지방산 분할에 대한 지방산결합 단백질 5의 역할)

  • Park, Jae-Seung
    • Journal of Digital Convergence
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    • v.17 no.8
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    • pp.283-291
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    • 2019
  • The aim of investigated the role of FABP5 in the hepatic lipogenesis and lipid metabolisms. Mice were overexpressed and silenced liver FABP5 using virus particles. Mice were fed a Western-type diet or regular chow for 1week and then sacrificed mouse after 24hr fasted. Liver homogenates were used for protein analysis by Western blot and mRNA levels by RT-PCR. Hepatic and serum lipids were analysed by thin-layer chromatography. Mice fed a Western-type or high saturated fat diet revealed large increases in FABP5 expression. However, FABP5 mRNA levels were drastically reduced under fasted. Hepatic TG was significantly increased FABP5-OEAV mice, but a significantly decreased hepatic free cholesterol under fed. The discovered a substantial decrease in hepatic TG mass with FABP5 silencing. In these data, presented evidence for an important role of FABP5 in hepatic lipogenesis and hepatic TG storage. FABP5 may also be a potential target in the treatment of NAFLD, metabolic syndrome, and obesity. Furthermore, studies to which transcription factors are involved in FABP5 expression and regulation.

Safety and Efficacy of Fowl Adenovirus Serotype-4 Inactivated Oil Emulsion Vaccine (닭 유래 아데노 바이러스 혈청형 4형(FAdV-4) 사독 오일 백신의 안전성 및 효능 평가)

  • Kim, Ji-Ye;Kim, Jong-Nyeo;Mo, In-Pil
    • Korean Journal of Poultry Science
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    • v.37 no.3
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    • pp.255-263
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    • 2010
  • Inclusion body hepatitis-hydropericardium syndrome (IBH-HPS) is an acute viral disease usually found in broilers aged from 3 to 5 weeks and causes up to 75% mortality. Among the 12 serotypes of fowl adenovirus group 1, serotype-4 (FAdV-4) was identified as a primary agent of IBH-HPS and was usually isolated in IBH-HPS cases in Korea since 2007. To prevent these IBH-HPS outbreaks in Korea, we developed the FAdV-4 inactivated vaccine using Korean isolate (ADL070244) and evaluated the efficacy of this vaccine. For the efficacy test, 2-week-old specific-pathogen-free (SPF) chickens intramuscularly inoculated with 1 or 2 dose of inactivated vaccine were used and challenged with FAdV-4 through either intramuscular or oral route at 2 weeks after vaccination. The vaccine induced good seroconversion which was confirmed by agar gel precipitation (AGP) test. In addition, the vaccine could decrease the FAdV-4 detection rate and histological lesion severity such as lymphocyte infiltration and necrosis in the liver comparing with those of non-vaccination group. Based on the current results, the developed FAdV-4 inactivated vaccine in this study was effective in the terms of reduction of virus detection rate and histological lesions severity. However, it was difficult to confirm the efficacy of the vaccine clearly because of no mortality and clinical signs in the non-vaccinated group after challenge. Therefore, we need further study to develop a standard challenged model system which could clearly evaluate the efficacy of the vaccines for FAdV-4.

EFFECTS OF SIGNAL TRANSDUCTION PATHWAY IN THE RAS-INDUCED CELLULAR TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (인체 상피세포에서 ras-종양유전자의 발암화가 신호 전달 기작에 미치는 영향)

  • Jang, Do-Geun;Byeon, Ki-Jeong;Kim, Chin-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.3
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    • pp.254-261
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    • 2000
  • The present study has attempted to look into the mechanism of ras-induced carcinogenesis in a human epithelial cell system. Human epithelial cells immortalized with Ad12-SV40 hybrid virus were used to assess carcinogenic potential of the ras-oncogene. Cells transfected with pSV2-ras showed characteristics of cellular transformation. The transformation parameters such as cell density, soft-agar colony formation, and cell aggregation were significantly increased in the cells expressing ras oncoprotein. In addition, the duration required for the appearance of foci was shortened in the ras-transfected cells. Consistent with other reports, our results demonstrated an evidence that the ras-oncogene induced the cellular transformation of human epithelial cell system. When a high concentration of glucocorticoid was added into the media, transformation process was accelerated. It is speculated that glucocorticoid may provide an advantageous environment for the proliferation of the transformed cells. The induction of the intracellular free calcium concentrations following agonist treatment was significantly lower in the transformed cells than in the control cells. These effects were more manifested in the presence of extracellular cacium, indicating that the transformation process may alter the influx pathway of extracellular calcium. The induction of $IP_3$ following agonist treatment was also lower in the transformed cells than in the control cells. Thus, it is suggested that phospholipase C-coupled pathway was down-regulated in the process of the ras-induced transformation. While the levels of $TGF-{\beta}_1$ and PAI-2 mRNAs were decreased, the level of fibronectin mRNA was increased. The results indicate that mechanism of the ras-induced transformation may be associated with the altered expressions of growth regulatory factors. The present study demonstrates an evidence that the ras-induced cellular transformation may be associated with alteration of signal transduction and growth regulatory factors. The study will contribute to improve the understanding of molecular mechanism of epithelium-derived cancers including oral cancer.

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The Effects of Nuclear Factor-κB Decoy Oligodeoxynucleotide on Lipopolysaccharide-Induced Direct Acute Lung Injury (리포다당질로 인한 직접성 급성폐손상에서 Nuclear Factor-κB Decoy Oligodeoxynucleotide의 효과)

  • Kim, Je Hyeong;Yoon, Dae Wui;Jung, Ki Hwan;Kim, Hye Ok;Ha, Eun Sil;Lee, Kyoung Ju;Hur, Gyu Young;Lee, Sung Yong;Lee, Sang Yeub;Shin, Chol;Shim, Jae Jeong;In, Kwang Ho;Yoo, Se Hwa;Kang, Kyung Ho
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.2
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    • pp.95-104
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    • 2009
  • Background: The pathophysiologic mechanisms of early acute lung injury (ALI) differ according to the type of primary insult. It is important to differentiate between direct and indirect pathophysiologic pathways, and this may influence the approach to treatment strategies. NF-$\kappa$B decoy oligodeoxynucleotide (ODN) is a useful tool for the blockade of the expression of NF-$\kappa$B-dependent proinflammatory mediators and has been reported to be effective in indirect ALI. The purpose of this study was to investigate the effect of NF-$\kappa$B decoy ODN in the lipopolysaccharide (LPS)-induced direct ALI model. Methods: Five-week-old specific pathogen-free male BALB/c mice were used for the experiment. In the preliminary studies, tumor necrosis factor (TNF)-$\alpha$, interleukine (IL)-6 and NF-$\kappa$B activity peaked at 6 hours after LPS administration. Myeloperoxidase (MPO) activity and ALI score were highest at 36 and 48 hours, respectively. Therefore, it was decided to measure each parameter at the time of its highest level. The study mice were randomly divided into three experimental groups: (1) control group which was administered 50 ${\mu}L$ of saline and treated with intratracheal administration of 200 ${\mu}L$ DW containing only hemagglutinating virus of Japan (HVJ) vector (n=24); (2) LPS group in which LPS-induced ALI mice were treated with intratracheal administration of 200 ${\mu}L$ DW containing only HVJ vector (n=24); (3) LPS+ODN group in which LPS-induced ALI mice were treated with intratracheal administration of 200 ${\mu}L$ DW containing 160 ${\mu}g$ of NF-$\kappa$B decoy ODN and HVJ vector (n=24). Each group was subdivided into four experimental subgroups: (1) tissue subgroup for histopathological examination for ALI at 48 hours (n=6); (2) 6-hour bronchoalveolar lavage (BAL) subgroup for measurement of TNF-$\alpha$ and IL-6 in BAL fluid (BALF) (n=6); (3) 36-hour BAL subgroup for MPO activity assays in BALF (n=6); and (4) tissue homogenate subgroup for measurement of NF-$\kappa$B activity in lung tissue homogenates at 6 hours (n=6). Results: NF-$\kappa$B decoy ODN treatment significantly decreased NF-$\kappa$B activity in lung tissues. However, it failed to improve the parameters of LPS-induced direct ALI, including the concentrations of tumor necrosis factor-$\alpha$ and interleukin-6 in BALF, myeloperoxidase activity in BALF and histopathologic changes measured by the ALI score. Conclusion: NF-$\kappa$B decoy ODN, which has been proven to be effective in indirect models, had no effect in the direct ALI model.

A Clinicopathological Study of Idiopathic Membranous Nephropathy in Children (소아에서 일차성 막성 신병증의 임상-병리학적 고찰)

  • Lee Bum-Hee;Cho Hee-Yeon;Kang Ju-Hyung;Kang Hee-Gyung;Ha Il-Soo;Cheong Hae-Il;Lee Hyun-Soon;Choi Yong
    • Childhood Kidney Diseases
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    • v.7 no.2
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    • pp.133-141
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    • 2003
  • Purpose : Idiopathic Membranous Nephropathy(IMN) is a rare renal disease in children. To help better understanding of its clinical course and treatment strategies, we reviewed the clinical manifestations and pathological findings of children with IMN. Methods : Among 58 cases with MN, from 1977 to 2003, 42(72.4%) were hepatitis B virus (HBV) associated and 16(27.6%), 6 males and 10 females, were idiopathic. All cases diagnosed aster 2000 were IMN. Several clinicopathological findings(sex, onset age, proteinuria, serum albumin, cholesterol, creatinine clearance, tubulointerstitial changes, glomerular sclerosis, hypertension, renal vein thrombosis, the use of ACE inhibitor, and immunosuppressive therapy) were compared between the remission and the non-remission group of the patients with IMN. Results : The median onset age was 13.4 years. Clinical manifestations were nephrotic syn-drome(7 cases, 43.8%), gross hematuria(5 cases, 31.3%) and microscopic hematuria with proteinuria(3 cases, 18.8%). Hypertension, hypocalcemic tetany and renal vein thrombosis were accompanied in 2, 1 and 2 cases, respectively. In addition to the typical findings of MN, the kidney biopsies showed segmental sclerosis(5 cases, 31.3%) or global sclerosis(6 cases, 37.5 %), diffuse crescents(1 case), and mild(11 cases, 68.7%) or moderate tubulointerstitial changes(3 cases, 18.8%). Thirteen cases(86.7%) received oral steroid. Among them 2 cases received cyclophophamide and 1 received cyclosporin as well. Ten cases(62.5%) received ACE inhibitors. In the patients followed up, 7 cases(46.7%) became free from proteinuria (remission group) while 8(53.3%) presented continous proteinuria (non-remission group), two (13.3%) of which progressed to renal failure. Clinicopathological findings showed no significant differences between the two groups. Conclusion : With HBV vaccination, HBV associated MN decreased markedly and IMN has taken up most of MN in children. For better understanding of this rare disease, a prospective multicenter study of the clinical course and treatment strategies should be done.

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Comparison of pathogen detection from wild and cultured olive flounder, red sea bream, black sea bream and black rockfish in the coastal area of Korea in 2010 (2010년 한국 연근해 자연산과 양식산 넙치, 참돔, 감성돔, 조피볼락의 병원체 비교)

  • Park, Myoung Ae;Do, Jeung-Wan;Kim, Myoung Sug;Kim, Seok-Ryel;Kwon, Mun-Gyeong;Seo, Jung Soo;Song, Junyoung;Choi, Hye-Sung
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.263-270
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    • 2012
  • This study surveyed for the prevalence of parasites, bacteria and viruses in four fish species, olive flounder (Paralichthys olivaceus), red sea bream (Pagrus major), black sea bream (Acathopagrus schlegeli) and black rockfish (Sebastes schlegeli) in 2010. The survey was aimed to compare the pathogens detected from wild and cultured fish for an epidemiological study. Anisakis sp. was predominantly detected from wild olive flounder and red sea bream (58.6% and 41.7% respectively), but not from the cultured fishes, suggesting anisakid infection is rare in cultured fish. The wild fish get in contact with the anisakids through their prey such as small fishes or crustaceans which carry the anisakids; whereas the cultured fish are fed with formulated feed, free of anisakids. Bacterial detection rates from the wild fishes examined in the study were lower than those of cultured fishes. Vibrio sp. dominated among detected bacterial population in cultured olive flounder (18%). Since vibriosis is known as a secondary infection caused by other stressful factors such as parasitic infections, handling and chemical treatment, it seems that cultured olive flounder are exposed to stressful environment. Viruses diagnosed in the study showed difference in distribution between wild and cultured fishes; hirame rhabdovirus (HRV) (0.1%) and lymphocystis disease virus (LCDV) (3.9%) were detected in the cultured olive flounder, but not in the wild fish, and marine birnavirus (MBV) (1.7%) and red sea bream iridovirus (RSIV) (3.2%) were detected from the wild and cultured red sea bream, respectively. From the survey conducted, it can be concluded that even though some pathogens (Trichodina sp., Microcotyle sp., etc.) are detected from both the wild and cultured fish, pathogens such as Anisakis sp., Vibrio sp. and LCDV showed difference in distribution in the wild and cultured host of same fish species and this can be attributed to their environmental condition and feeding.

Genetic Characterization of Antigenic Variant Infectious Bursal Disease Virus (IBDV) in Chickens in Korea

  • Jong-Yeol Park;Ki-Woong Kim;Ke Shang;Sang-Won Kim;Yu-Ri Choi;Cheng-Dong Yu;Ji-Eun Son;Gyeong-Jun Kim;Won-Bin Jeon;In-Hwan Kim;Bai Wei;Min Kang;Hyung-Kwan Jang;Se-Yeoun Cha
    • Korean Journal of Poultry Science
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    • v.50 no.4
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    • pp.231-240
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    • 2023
  • Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive disease in young chickens, and causes considerable economic losses to the poultry industry. More than 30 years ago, an antigenic variant IBDV (avIBDV) was reported in chicken farms in the United States. Recently, a novel avIBDV exhibited clear differences in molecular characteristics compared with previous variant strains. This study investigated the molecular characteristics of recently isolated avIBDV strains in Korea. Strains of avIBDV were confirmed by reverse transcription PCR (RT-PCR) and were propagated in 10-day-old specific-pathogen-free (SPF) embryonated chicken eggs through chorioallantoic membrane (CAM) inoculation. Multiple sequence alignment and phylogenetic analyses of hypervariable regions VP2 gene revealed that the strains originated from two different avIBDV lineages (G2a and G2d). In our results, we confirmed the co-existence and prevalence of avIBDV genogroup G2a and G2d in chicken farms. It is necessary to study the protective efficacy of current vaccines against avIBDVs.

A study on the classification of research topics based on COVID-19 academic research using Topic modeling (토픽모델링을 활용한 COVID-19 학술 연구 기반 연구 주제 분류에 관한 연구)

  • Yoo, So-yeon;Lim, Gyoo-gun
    • Journal of Intelligence and Information Systems
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    • v.28 no.1
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    • pp.155-174
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    • 2022
  • From January 2020 to October 2021, more than 500,000 academic studies related to COVID-19 (Coronavirus-2, a fatal respiratory syndrome) have been published. The rapid increase in the number of papers related to COVID-19 is putting time and technical constraints on healthcare professionals and policy makers to quickly find important research. Therefore, in this study, we propose a method of extracting useful information from text data of extensive literature using LDA and Word2vec algorithm. Papers related to keywords to be searched were extracted from papers related to COVID-19, and detailed topics were identified. The data used the CORD-19 data set on Kaggle, a free academic resource prepared by major research groups and the White House to respond to the COVID-19 pandemic, updated weekly. The research methods are divided into two main categories. First, 41,062 articles were collected through data filtering and pre-processing of the abstracts of 47,110 academic papers including full text. For this purpose, the number of publications related to COVID-19 by year was analyzed through exploratory data analysis using a Python program, and the top 10 journals under active research were identified. LDA and Word2vec algorithm were used to derive research topics related to COVID-19, and after analyzing related words, similarity was measured. Second, papers containing 'vaccine' and 'treatment' were extracted from among the topics derived from all papers, and a total of 4,555 papers related to 'vaccine' and 5,971 papers related to 'treatment' were extracted. did For each collected paper, detailed topics were analyzed using LDA and Word2vec algorithms, and a clustering method through PCA dimension reduction was applied to visualize groups of papers with similar themes using the t-SNE algorithm. A noteworthy point from the results of this study is that the topics that were not derived from the topics derived for all papers being researched in relation to COVID-19 (

    ) were the topic modeling results for each research topic (
    ) was found to be derived from For example, as a result of topic modeling for papers related to 'vaccine', a new topic titled Topic 05 'neutralizing antibodies' was extracted. A neutralizing antibody is an antibody that protects cells from infection when a virus enters the body, and is said to play an important role in the production of therapeutic agents and vaccine development. In addition, as a result of extracting topics from papers related to 'treatment', a new topic called Topic 05 'cytokine' was discovered. A cytokine storm is when the immune cells of our body do not defend against attacks, but attack normal cells. Hidden topics that could not be found for the entire thesis were classified according to keywords, and topic modeling was performed to find detailed topics. In this study, we proposed a method of extracting topics from a large amount of literature using the LDA algorithm and extracting similar words using the Skip-gram method that predicts the similar words as the central word among the Word2vec models. The combination of the LDA model and the Word2vec model tried to show better performance by identifying the relationship between the document and the LDA subject and the relationship between the Word2vec document. In addition, as a clustering method through PCA dimension reduction, a method for intuitively classifying documents by using the t-SNE technique to classify documents with similar themes and forming groups into a structured organization of documents was presented. In a situation where the efforts of many researchers to overcome COVID-19 cannot keep up with the rapid publication of academic papers related to COVID-19, it will reduce the precious time and effort of healthcare professionals and policy makers, and rapidly gain new insights. We hope to help you get It is also expected to be used as basic data for researchers to explore new research directions.


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