• Title/Summary/Keyword: Molecular Characteristics

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Morphological Changes of Bones and Joints with Rheumatoid Arthritis and Osteoarthritis

  • Hong, Yun-Kyung;Javaregowda, Palaksha Kanive;Lee, Sang-Kil;Lee, Sang-Rae;Chang, Kyu-Tae;Hong, Yong-Geun
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.143-149
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    • 2011
  • Arthritis is a common disease in aged people, and is clinically divided into rheumatoid arthritis (RA) and osteoarthritis (OA). Although common symptoms such as pain are present, the underlying pathological mechanisms are slightly different. Therefore, the objectives of the present study were to compare joint damage induced by RA and OA by analyzing the major morphological and molecular differences, and to propose a suitable therapeutic intervention based on the pathophysiological conditions of bones and joints. For the RA animal model, 8-week-old DBA1/J mice were immunized with bovine type II collagen emulsified in complete Freund's adjuvant (CFA). Normal C57BL/6 mice (over 2 years of age) were used for OA. The clinical arthritis score was calculated using a subjective scoring system, and paw thicknesses were measured using calipers. The serum TNF ${\alpha}$ level was analyzed using an ELISA kit. Micro-CT was used to identify pathological characteristics and morphological changes. In collagen-induced RA mice, there were increased ankle joint volumes and clinical scores (p<0.01). The concentration of TNF ${\alpha}$ was significantly increased from 3 to 7 weeks after immunization. Micro-CT images showed trabecular bone destruction, pannus formation, and subchondral region destruction in RA mice. OA among aged mice showed narrowed joint spaces and breakdown of articular cartilage. This study suggests that a careful therapeutic intervention between RA and OA is required, and it should be based on morphological alteration of bone and joint.

Comparison of recovery rates of DNPH-Cartridge sampler in the analysis of odorous carbonyl compounds (DNPH Cartridge를 이용한 악취성 카르보닐화합물의 분석회수율 비교)

  • Hong, One-Feel;Lim, Sung-Min;Kim, Ki-Hyun
    • Analytical Science and Technology
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    • v.23 no.5
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    • pp.446-456
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    • 2010
  • In this study, the performance characteristics of DNPH sampling were investigated in the collection and analysis of 5 carbonyl compounds (CC) in air using the cartridge products produced by three different makers. For these experiments, gaseous standards of 5 CCs were prepared to cover 9 concentration levels for each compound (33~2600 nmol). Some cartridge products exhibited relatively high blank values of acetaldehyde (AA) and propionaldehyde (PA). The recovery rates of all three cartridges showed moderate reduction as the molecular weight of CC increased. In addition, when the recovery rate was compared by percent error (%), the most stable patterns were achieved in the intermediate concentration range of 263~1312 nmol (in case of AA). The overall results of our study suggest that the optimal range of recovery for a given concentration range should be considered to obtain the most reliable data for the DNPH cartridge method.

Platform Technology for Food-Grade Expression System Using the genus Bifidobacterium

  • Park, Myeong-Soo;Kang, Yoon-Hee;Cho, Sang-Hee;Seo, Jeong-Min;Ji, Geun-Eog
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.155-157
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    • 2001
  • Bifidobacterium spp. is nonpathogenic, gram-positive and anaerobic bacteria, which inhabit the intestinal tract of humans and animals. In breast-fed infants, bifidobacteria comprise morethan 90% of the gut bacterial population. Bifidobacteria spp. are used in commericial fermented dairy products and have been suggested to exert health promoting effects on the host by maintaining intestinal microflora balances, improving lactose tolerance, reducing serum cholesterol levels, increasing synthesis of vitamins, and aiding the immune enchancement and anticarcinogenic activity for the host. These beneficial effects of Bifidobacterium are strain-related. Therefore continued efforts to improve strain characteristics are warranted. in these respect, development of vector system for Bifidobacterium is very important not only for the strain improvement but also because Bifidobacterium is most promising in serving as a delivery system for the useful gene products, such as vaccine or anticarcinogenic polypeptides, into human intestinal tract. For developing vector system, we have characterized several bifidobacterial plasmids at genetic level and developed several shuttle vectors between E. coli and Bifidobacterium using them. Also, we have cloned and sequenced several metabolic genes and food grade selection marker. Also we have obtained bifidobacterial surface protein, which will be used as the mediator for surface display of foreign genes. Recently we have succeeded in expressing amylase and GFP in Bifidobacterium using our own expression vector system. Now we are in a very exciting stage for the molecular breeding and safe delivery system using probiotic Bifidobacterium strains.

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Purification and Characterization of the Fibrinolytic Enzyme Produced by Bacillus subtilis KCK-7 from Chungkookjang

  • Paik, Hyun-Dong;Lee, Si-Kyung;Heo, Seok;Kim, Soo-Young;Lee, Hyung-Hoan;Kwon, Tae-Jong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.829-835
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    • 2004
  • A fibrinolytic enzyme has been found in several bacteria isolated from fermented food. This study was carried out to investigate the purification and characteristics of the fibrinolytic enzyme produced by Bacillus subtilis KCK-7 originated from Chungkookjang. The fibrinolytic enzyme was purified to homogeneity from the culture supernatant using ammonium sulfate fractionation and chromatographies on DEAE-cellulose and on Sephadex G-100. The final specific activity of the purified enzyme increased 11.0-fold, and the protein amount in the purified enzyme was about 16% of that in the culture supernatant. The molecular weight of the purified enzyme was estimated to be about 45,000 by SDS-PAGE. The optimum pH and temperature for the enzyme activity were pH 7.0 and $60^{\circ}C$, respectively. The enzyme activity was relatively stable up to $60^{\circ}C$ over the pH range of 7.0-10.0. The fibrinolytic enzyme activity increased by $Ca^{2+}$ and $Cu^{2+}$, whereas it was inhibited by $Hg^{2+}$ and $Ba^{2+}$. In addition, it was severely inhibited by PMSF and DFT. It is suggested that the purified enzyme was a serine protease for the fibrinolysis. The purified enzyme could completely hydrolyze fibrin in vitro within 8 h. Hence, it is suggested that the purified enzyme can be put into practice as an effective thrombolytic agent.

Analysis of Squalene Synthase Expression During the Development of Ganoderma lucidum

  • Zhao, M.W.;Zhong, J.Y.;Liang, W.Q.;Wang, N.;Chen, M.J.;Zhang, D.B.;Pan, Y.J.;Jong.S.C.
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.116-120
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    • 2004
  • The medicinal properties of Ganoderma lucidum have been recognized in China for many centuries. Active pharmaceutical components include triterpenes. To elucidate the molecular regulation of triterpene biosynthesis in this mushroom, a 57-base pair DNA fragment encoding the fourth conserved domain SQ-4 (SMGLFLQKTNIIRDYNEDL) of squalene synthase was synthesized and cloned into the expression vector pET-32a(+). The recombinant fusion protein induced by IPTG (isopropyl-$\beta$-D-thiogalactopyranoside) was overexpressed in the Escherichia coli. Using the purified recombinant fusion protein of 20.9 kDa, a specific polyclonal antibody was obtained from immunized rabbit. Expression of squalene synthase at different development stages of Ganoderma lucidum was analyzed.

Studies on the Development and the Characteristics of the Powerful Raw Starch Digesting Enzyme (강력한 생전분 분해효소의 개발과 특성)

  • ;;Hajime Taniguchi
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.251-259
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    • 1990
  • Asp. usumii IAM 2185 was selected as a strain producing the powerful raw starch digesting glucoamylase. The optimum initial pH, the optimum temperature and the optimum cultural time for the enzyme production on wheat bran medium were pH 6-8,25-$30^{\circ}C$ and 72 hrs, respectively. The addition of ammonium nitrate and albumin on wheat bran medium, respectively, increase slightly the enzyme production. The enzyme was purified by ammonium sulfate fractionation, CM-cellulose and DEAE-cellulose column chromatography. The specific activity of the purified enzyme was 34.3 U/mg protein and the yield of enzyme activity was 10.3%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis and its molecular weight was estimated to be 67,000 by SDS polyacrylamide disc gel electrophoresis. The isoelectric point for the purified enzyme was pR 3.7. The optimum temperature and optimum pH were $60^{\circ}C$and pH 3.0 and the purified enzyme was stable in the pH range of 1.0-11.0. The purified enzyme was stable below $50^{\circ}C$ and its thermostability was greatly increased by the addition of $Ca^{2+}$. The purified enzyme showed a high hydrolysis rate on various raw starches such as corn, rice, yam, arrow root, sweet potato and glutinous rice.

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Conditions for Transformation of Alkalophilic Bacillus sp. K-17 (호알칼리성 Bacillus속 B-17의 형질전환조건)

  • 성낙계;정운상;고학룡;정정희
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.213-218
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    • 1989
  • To investigate the possibility of using alkalophilic Bacillus sp. K-11 as a host for molecular cloning, plasmid pUB110 and pBD64 were introduced into alkalophilic Bacillus sp. K-17 by protoplast transformation system. Protoplasts of Bacillus sp. K-11 were prepared by treatment with 200 $\mu\textrm{g}$/$m\ell$ Iysozyme in SMM buffer containing 0.4M sucrose. Optimal temperature, pH and culture time for protoplast formation were 4$0^{\circ}C$, 7.0 and 4hrs, respectively. Cell wall was regenerated efficiently on DM3 medium containing 0.8% agar and 0.5M sodium succinate. Under these conditions for protoplast formation and regeneration, the highest transformation efficiency was obtained with cells incubated for 4hrs, and using 30%(V/V) of 40%(W/V) PEG6,000, In characteristics of transfer-mants, plasmid pUB110 was more stable than plasmid pBD64 in Bacillus sp. K-17. Maximum xylanase production of both transformants carrying pUB110 and pBD64, respectively was similar, but under the same conditions, enzyme secretion by transformant carrying pUB110 was earlier than that of transformant carrying pBD64.

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Molecular cloning of metal-responsive transcription factor-1 (MTF-1) and transcriptional responses to metal and heat stresses in Pacific abalone, Haliotis discus hannai

  • Lee, Sang Yoon;Nam, Yoon Kwon
    • Fisheries and Aquatic Sciences
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    • v.20 no.7
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    • pp.9.1-9.13
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    • 2017
  • Background: Metal-responsive transcription factor-1 (MTF-1) is a key transcriptional regulator playing crucial roles in metal homeostasis and cellular adaptation to diverse oxidative stresses. In order to understand cellular pathways associated with metal regulation and stress responses in Pacific abalone (Haliotis discus hannai), this study was aimed to isolate the genetic determinant of abalone MTF-1 and to examine its expression characteristics under basal and experimentally stimulated conditions. Results: The abalone MTF-1 shared conserved features in zinc-finger DNA binding domain with its orthologs; however, it represented a non-conservative shape in presumed transactivation domain region with the lack of typical motifs for nuclear export signal (NES) and Cys-cluster. Abalone MTF-1 promoter exhibited various transcription factor binding motifs that would be potentially related with metal regulation, stress responses, and development. The highest messenger RNA (mRNA) expression level of MTF-1 was observed in the testes, and MTF-1 transcripts were detected during the entire period of embryonic and early ontogenic developments. Abalone MTF-1 was found to be Cd inducible and highly modulated by heat shock treatment. Conclusion: Abalone MTF-1 possesses a non-consensus structure of activation domains and represents distinct features for its activation mechanism in response to metal overload and heat stress. The activation mechanism of abalone MTF-1 might include both indirect zinc sensing and direct de novo synthesis of transcripts. Taken together, results from this study could be a useful basis for future researches on stress physiology of this abalone species, particularly with regard to heavy metal detoxification and thermal adaptation.

Stress Analysis of Total Knee System Depending on Implant Materials and Fixation Methods (인공무릎관절에 있어서 임플란트의 재료 및 고정방법에 따른 응력분석)

  • Cho, C.H.;Cho, Y.K.;Choi, J.B.;Choi, K.
    • Proceedings of the KOSOMBE Conference
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    • v.1997 no.11
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    • pp.484-488
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    • 1997
  • Three-dimensional finite element analyses were used to compare the stress distribution and the stability of the fixation among seven different tibial components and to investigate the effect due to implant materials in total knee arthroplasty. The components included an intact tibia(Type I), Cemented Cobalt-Chromium tibial tray implanted with a PMMA cemented Co-Cr stem(Type II), Cemented Co-Cr tibial tray with a uncemented Co-Cr stem(Type III), Cemented Ultra High Molecular Weight Polyethylene (UHMWPE) tibial tray with a cemented UHMWPE stem (Type IV), Cemented UHMWPE tray with a uncemented UHMWPE stem(Type V), Cemented Co-Cr tray without a stem(Type VI), and Cemented UHMWPE tray without a stem(Type VII). Uncemented components were assumed to have complete bony in growth and a rigid state of fixation between component and bone. The interface between bone/cement/component of cemented components was also assumed to be fully bonded. Bi-condylar forces were applied. The results indicated that Uncemented stem components provided lower bone stress shielding and stress concentration. The UHMWPE tray and stem component showed better agreement with the intact tibia than the Co-Cr Alloy tray and stem components. If the implant tray can be fixed firmed without a stem, Cemented PE tray without a stem(Type VII) may be recommended to give the best characteristics in the sense of stress distribution and stability.

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Biovars of Pseudomonas syringae pv. actinidiae Strains, the Causal Agent of Bacterial Canker of Kiwifruit, Isolated in Korea (우리나라에서 분리한 참다래 궤양병균 Pseudomonas syringae pv. actinidiae 균주들의 Biovar)

  • Lee, Young Sun;Kim, Jin;Kim, Gyoung Hee;Choi, Eu Ddeum;Koh, Young Jin;Jung, Jae Sung
    • Research in Plant Disease
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    • v.23 no.1
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    • pp.35-41
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    • 2017
  • Pseudomonas syringae pv. actinidiae is the causative agent of bacterial canker of kiwifruit. The population of this pathogen is differentiated into three biovars, biovar 1, 2 and 3, according to their molecular characteristics. In this work, we determined biovars of P. syringae pv. actinidiae strains isolated in Korea since 1997 and stored in Department of Biology, Sunchon National University, Suncheon, Korea. The biovars of P. syringae pv. actinidiae strains were determined by PCR using biovar specific primers developed previously. Of 682 strains investigated, 288 strains belonged to biovar 2, while 394 strains were biovar 3. There were no P. syringae pv. actinidiae strains belonging to biovar 1 among the strains isolated in Korea. Sudden outbreak and spreading of bacterial canker caused by biovar 3 strain suggest that this strain has character of rapid transmission.