• Title/Summary/Keyword: 2-D Electrophoresis

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Systematic Study on the Fishes of the Family Cobitidae(Pisces, Cypriniformes). 2. Taxonomic Study on the Cobitis taenia complex from Korea (기름종개과(family Cobitidae)어류의 계통분류에 관한 연구 2. 한국산 Cobitis taenia complex의 분류학적 고찰)

  • 양서영;이혜영;양홍준;전상린;박병상;김재흡
    • Animal Systematics, Evolution and Diversity
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    • v.9 no.2
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    • pp.151-170
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    • 1993
  • Morphometric, band-pattern and electrophoretic analysis on Cobitis taenia complex were performed to investigate the morphological and genetic differentiation and to clarify their taxonomic status. Intermediate types of band-pattern (C and D type) were more frequently expressed than that of types of C. t. taenia(type A) and C. t. lutheri (type B). Sexual dimorphism of band-pattern was observed not only in C. t. taenia(type A) and C. t. lutheri(type B). Sexual dimorphism of band-pattern was observed not only in C. t. lutheri but also in C. t. taenia and C. t. striata as well. Discriminant function analysis based on 19 morphological characters shows no significant differences among C. taenia complex. The degree of genic variation of C. t. striata was higher ( =1.48, P=31.2%, HD=0.009) than those of C. t. striata was higher( =1.48, P=31.2%, HD=0.082 and HG=0.009) than those of C. t. lutheri ( =1.37, P=2.7%, HD=0.058 and HG=0.065). The average genetic similarities between C. t. lutheri and C. t. taenia-C. t. striata were S=0.62 and S=0.66 respectively and these values indicate that C. t. tanenia has evolved specific level of differentiation. C. t. striata and C. t. lutheri show subspecifc level of close genetic similarity (S=0.82). Based on the divergent time estimate (Nei, 1975) it is assumed that C. t. tanenia was branched off from the other subspecies about two million years before present (MYBP) and C. t. striata and C. t. lutheri were differentiated about 0.6 MYBP. The use of C. sinesis an the scientific name for the Korean C. t. taenia, proposed by Kim and Lee (1988) seems incorrect since they are quite different in the structure of lamina circularis (Vladycov, 1935), the external morphology and distribution (Cheng and Zheng, 1987) and the chromosome number(Yu et al., 1989). Kim and Lee(1988) also argued that C.t. striata and C. t. lutheri should be treated as distinct species but the present study and other reports (Kim and Lee, 1984; Kim and Yang, 1993) do not support it. We conclude that C. t. taenia is a good species and C. t. striata and C. t. lutheri are subspecific status. Their scientific names should be revised in the future.

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Immuno-stimulating Activities of Polysaccharides Isolated from Commercial Soy Sauce and Traditional Korean Soy Sauce (시판양조 및 재래식 조선간장으로부터 분리한 다당의 면역증강 활성 비교)

  • Park, Hye-Ryung;Lee, Moon-Su;Jo, Sun-Young;Won, Hye-Jin;Lee, Hyun-Sun;Lee, Ho;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.44 no.2
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    • pp.228-234
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    • 2012
  • The varying characteristics between traditional and commercial soy sauce may be initiated by raw materials and fermentation techniques for the production of $meju$ and $koji$. We examined properties regarding polysaccharides isolated from commercial soy sauce made by the $koji$ process (CSP-0) and Korean traditional soy sauce made by the $meju$ process (KTSP-0) as well as their immuno-stimulating activities. KTSP-0 had rhamnogalacturonan II (RG-II) including 1.1% of unusual monosaccharides 3-deoxy-D-$manno$-2-octulosonic acid (KDO). Anti-complementary activities of CSP-0 and KTSP- 0 were increased dose-dependently but KTSP-0 (64.7%) was higher than CSP-0 (56%) at $1,000{\mu}g/mL$. C3 activation products were identified by crossed immuno-electrophoresis. CSP-0 caused complementary activations $via$ only classical pathway while KTSP-0 caused complementary activations $via$ both alternative and classical pathways. KTSP-0 significantly increased the secretion of interleukin (IL)-6 at $8-1,000{\mu}g/mL$ and IL-12 at $40{\mu}g/mL$ on macrophages. The results suggest that the immuno-stimulating activity of KTSP-0 is greater than that of CSP-0 from anti-complementary activity.

Identification of Novel Bacillus subtilis IDCC 9204 Producing a High-Level Fibrinolytic Enzyme and Properties of NK-IL9204 (고농도 혈전용해효소를 생산하는 신규 Bacillus subtilis IDCC 9204의 분리 및 NK-IL9204의 효소학적 특성)

  • Lee, Seung-Hun;An, Gwangmin;Kim, Heu-Hang;Kang, Jae-Hoon;Kang, Dae-Jung
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.600-606
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    • 2012
  • A Bacillus sp. that produces fibrinolytic enzyme was isolated from Cheonggukjang, a traditional Korean soybean-fermented food. According to 16S rRNA gene base sequencing, the bacillus was identified as a variety of Bacillus subtilis, and named Bacillus subtilis IDCC 9204. Fibrinolytic enzyme NK-IL9204 was stable up to $60^{\circ}C$ and within pH range of 5-10. Purified NK-IL9204 was detected through fibrin zymography. The molecular weight and isoelectric point of the enzyme were estimated to be 27.7 kDa and 6.7 by SDS-PAGE and 2D electrophoresis, respectively. Its amino acid sequence was similar to that of nattokinase (identities 99.5%) and different from that of nattokinase BPN (identities 86.4%). The plasma fibrinolytic activity of NK-IL9204 was measured by euglobulin clot lysis times (ECLT). The NK-IL9204 was orally administered to SD rats for 3 weeks (1,000 FU/rat/day). The ECLT was significantly shortened by supplementation of NK-IL9204.

Electrophoretic Analysis on the Protein Alteration in the Brain of Actylamide Administered Mouse (Acrylamide 에 의한 생쥐 뇌단백질의 변화양상에 관한 전기영동적 분석)

  • 김동수;하재청
    • The Korean Journal of Zoology
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    • v.33 no.4
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    • pp.461-467
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    • 1990
  • To investigate the neurological effect of acrylamide, whole brain of Intoxicated mouse induced early hindlimbs ataxia was examined by using the methods of SDS-PAGE and two-dimensional electrophoresis. In the gel patterns by SDS-PAGE, when the patterns of each group were compared relatively, there were no remakable changes but in the patterns of 2D-PAGE, some protein alterations were observed. Especially, the spots containing 20 (14,500, 5.64) and 21 (19,900, 6.78) were disappeared, and the spots 9 (31,300, 5.82), 11 (31,300, 5.36) and 19 (16,400, 5.42) showed marked decrease relatively in the case of treatment group. Among these changed spots, the spot 20 (14,500, 5.64) showed higher quantity than that of control group but several spots containing the spots 11 (31,300, 5.36), and 19(16.400, 5.42) were identical or equal to those of control In quantity in the case of recovery group. It seems that acrylamide might already inhibit the brain protein synthesis mechanism at the time of onset of distal neuropathy by participation in the protein metabolism so as to impair the brain regulation ability followed by a malfunction of mouse central nervous system (CNS) and recovery is gradually progressed with the dose and duration dependence after the cessation of acrylamide administration.

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Association of Succinate Dehydrogenase and Triose Phosphate Isomerase Gene Expression with Intramuscular Fat Content in Loin Muscle of Korean (Hanwoo) Cattle (한우 등심조직 내 succinate dehydrogenase 및 triosephosphate isomerase 발현이 근내 지방함량에 미치는 영향에 관한 연구)

  • Kim, Nam-Kuk;Lee, Seung-Hwan;Lim, Da-Jeong;Yoon, Du-Hak;Lee, Chang-Soo;Kim, Oun-Hyun;Kim, Hyeong-Cheol;Oh, Sung-Jong;Hong, Seong-Koo
    • Journal of Life Science
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    • v.22 no.1
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    • pp.31-35
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    • 2012
  • In a previous study, succinate dehydrogenase (SDH) and triose phosphate isomerase (TPI) were detected by 2D gel electrophoresis as differentially expressed proteins in the longissimus thoracis muscles of cattle aged between 12 and 27 months old. In the present study, we investigated the association of SDH and TPI gene expression with intramuscular fat content in 50 Hanwoo steers. The SDH gene was expressed at a 4 times higher level in the 12 month old group than in the 27 month old group (p<0.01). A regression analysis between gene expression value and intramuscular fat content showed a negative correlation between expression of the SDH gene and intramuscular fat content (p<0.001). In contrast, the expression of the TPI gene showed no significant association with intramuscular fat content. This result suggests that the SDH gene may be a candidate marker gene for intramuscular fat content in the longissimus thoracis of Korean cattle.

Serum Anti-Gal-3 Autoantibody is a Predictive Marker of the Efficacy of Platinum-Based Chemotherapy against Pulmonary Adenocarcinoma

  • Yanagita, Kengo;Nagashio, Ryo;Ryuge, Shinichiro;Katono, Ken;Jiang, Shi-Xu;Tsuchiya, Benio;Nakashima, Hiroyasu;Fukuda, Eriko;Goshima, Naoki;Saegusa, Makoto;Satoh, Yukitoshi;Masuda, Noriyuki;Sato, Yuichi
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7959-7965
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    • 2015
  • Background: Identification of predictive markers for the efficacy of platinum-based chemotherapy is necessary to improve the quality of the life of cancer patients. Materials and Methods: We detected proteins recognized by autoantibodies in pretreated sera from patients with lung adenocarcinoma (AC) evaluated as showing progressive disease (PD) or a partial response (PR) after cisplatin-based chemotherapy by proteomic analysis. Then, the levels of the candidate autoantibodies in the pretreated serum were validated by dot-blot analysis for 22 AC patients who received platinum-based chemotherapy, and the expression of identified proteins was immunohistochemically analyzed in 40 AC biopsy specimens. Results: An autoantibody against galectin-3 (Gal-3) was detected in pretreated sera from an AC patient with PD. Serum IgG levels of anti-Gal-3 autoantibody were significantly higher in patients evaluated with PD than in those with PR and stable disease (SD) (p = 0.0084). Furthermore, pretreated biopsy specimens taken from patients evaluated as showing PD following platinumbased chemotherapy showed a tendency to have a higher positive rate of Gal-3 than those with PR and SD (p = 0.0601). Conclusions: These results suggest that serum IgG levels of anti-Gal-3 autoantibody may be useful to predict the efficacy of platinum-based chemotherapy for patients with lung AC.

Proteomic Analysis and the Antimetastatic Effect of N-(4methyl)phenyl-O-(4-methoxy) phenyl-thionocarbamate-Induced Apoptosis in Human Melanoma SK-MEL-28 cells

  • Choi Su-La;Choi Yun-Sil;Kim Young-Kwan;Sung Nack-Do;Kho Chang-Won;Park Byong-Chul;Kim Eun-Mi;Lee Jung-Hyung;Kim Kyung-Mee;Kim Min-Yung;Myung Pyung-Keun
    • Archives of Pharmacal Research
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    • v.29 no.3
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    • pp.224-234
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    • 2006
  • We employed human SK-MEL-28 cells as a model system to identify cellular proteins that accompany N-(4-methyl)phenyl-O-(4-methoxy)phenyl-thionocarbamate (MMTC)-induced apoptosis based on a proteomic approach. Cell viability tests revealed that SK-MEL-28 skin cancer cells underwent more cell death than normal HaCaT cells in a dose-dependent manner after treatment with MMTC. Two-dimensional electrophoresis in conjunction with matrixassisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analysis or computer matching with a protein database further revealed that the MMTC-induced apoptosis is accompanied by increased levels of caspase-1, checkpoint suppressor-1, caspase-4, NF-kB inhibitor, AP-2, c-Jun-N-terminal kinase, melanoma inhibitor, granzyme K, G1/S specific cyclin D3, cystein rich protein, Ras-related protein Rab-37 or Ras-related protein Rab-13, and reduced levels of EMS (oncogene), ATP synthase, tyrosine-phosphatase, Cdc25c, 14-3-3 protein or specific structure of nuclear receptor. The migration suppressing effect of MMTC on SK-MEL-28 cell was tested. MMTC suppressed the metastasis of SK-MEL-8 cells. It was also identified that MMTC had little angiogenic effect because it did not suppress the proliferation of HUVEC cell line. These results suggest that MMTC is a novel chemotherapeutic and metastatic agents against the SK-MEL-28 human melanoma cell line.

Synthesis and Characterization of Polyethylenimine-conjugated Polydiacetylene Liposome as a Gene Delivery Carrier (폴리디아세틸렌 리포좀 표면에 저분자량의 폴리에틸렌이민을 연결한 새로운 유전자 전달체 합성 및 특징 연구)

  • Lee, Young Hwa;Yim, Kang Hyuck;Heo, Jungseok;Choi, Joon Sig
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.43-48
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    • 2014
  • In this paper, we made a new polycationic polymeric liposome composed of low molecular weight polyethylenimine (PEI) and 10,12-pentacosadiynoic acid (PCDA). PCDA liposome was prepared by ultraviolet irradiation. PEI was further conjugated on the surface of the polymerized PCDA liposome using coupling reagents to make PCDA-PEI. The blue-to-red transition of PCDA liposome was observed during the coupling reaction. The size distribution of liposome and complexes with plasmid DNA was measured by dynamic light scattering (DLS). The complex formation was also identified by agarose gel electrophoresis and PicoGreen reagent assay. We confirmed the complex formation of the polymeric liposome with DNA and then performed transfection and cytotoxicity assay in HEK 293 and HeLa cells. As a result, PCDA-PEI showed significant gene transfection efficiency and low cytotoxicity. This study shows that PEI-conjugated PCDA liposome could be an efficient gene or drug delivery carrier.

Genetic Properties and Antimicrobial Resistance of Campylobacter jejuni Isolates from Diarrhea Patients in Gyeonggi-do (경기도내에서 분리한 캠필로박터 제주니균의 유전적특성 및 항생제내성 연구)

  • Hur, Eun-Seon;Park, Po-Hyun;Kim, Jong-Hwa;Son, Jong-Sung;Yun, Hee-Jeong;Lee, Yea-Eun;Choi, Yun-Sook;Yoon, Mi-Hye;Lee, Jong-Bok
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.228-236
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    • 2013
  • Campylobacter jejuni is one of important food-borne pathogens causing human gastroenteritis. We isolated 42 strains of C. jejuni from diarrhea patients and 4 food-poisoning outbreaks in 2010, Gyeonggi-do. In this study, 42 strains were tested for genetic characteristics, the serotype distribution and antimicrobial resistant rate. The presence of hipO (100%), cdtB (100%), and mutated gyrA (95.2%) genes was detected in C. jejuni by polymerase chain reaction (PCR). Detection of mutated gyrA gene correlated with ciprofloxacin resistance. Forty isolates had mutated gyrA gene and were actually resistant to ciprofloxacin. Furthermore, comparing the gyrA DNA sequence data, ciprofloxacin-resistant isolates had a mutation of the DNA sequence from ACA (threonine) to ATA (isoleucine). But 41 strains (97.6%) of patient isolates were susceptible to erythromycin and azithromycin. A total of 35.7% among 42 C. jejuni isolates were identified into 4 different serotypes. The serotype distribution of C. jejuni strains were shown to be HS2(B), HS3(C), HS4(D), HS19(O). To investigate the genotypes of C. jejuni isolated in Gyeonggi province, repetitive sequence polymerase chain reaction (rep-PCR) analysis and SmaI-digested pulsed-filed gel electrophoresis (PFGE) profile analysis were performed. From the PFGE analysis of 42 C. jejuni strains, 12 clusters of PFGE profile were obtained. On the other hand, 11 clusters of rep-PCR profile were obtained from 42 strains of C. jejuni.

Postmortem Changes of the Protein and Amino Acid Composition of Muscles in the Partially Frozen Prawn, Pandalus japonica (보리새우육의 부분동결저장중 단백질 및 아미노산의 조성변화)

  • PYEUN Jae-Hyeung;CHOI Young-Joon;KIM Jeung-Han;CHO Kweon-Ock
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.280-290
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    • 1984
  • An extensive study has been made on the relationship between the freshness and the compositions of the muscle protein of prawn, Pandalus japonica during the storage under partially frozen condition. The variations of the subunit distribution for sarcoplasmic protein and myofibrillar protein extracted from the samples by changes of freshness were discussed by sodium dodecylsulfate-poly-acrylamide gel (SDS-PAG) electrophoresis. On the other hand, the denaturation constant ($K_D$) of the myofibrillar protein extracted from the prawn stored at $-3^{\circ}C\;and\;-20^{\circ}C$ were successively compared. The prawn muscle contained about $18\%$ of protein with the composition of $32\%$ in sarcoplasmic protein, $56\%$ in myofibrillar protein, $10\%$ in residual intracellular protein and $2\%$ in stroma. The indices for estimating freshness of the muscle were approached to the early stage of putrefaction on the 26th day of the storage with $25.29mg\%$ of total volatile basic nitrogen, $31.36\%$ of K-value and 8.83 of pH. The content of the myofibrillar protein was remarkably decreased with the time during the storage while that of residual intracellular protein was increased. The $K_D$ values of the myofibrillar protein were $9.03{\times}10^{-6}sec^{-1}\;at\;-3^{\circ}C\;and\;4.42{\times}10^{-6}sec^{-1}\;at\;-20^{\circ}C$. The results of the analysis of SDS-PAG electrophoretograms indicated that the sarcoplasmic protein and the myofibrillar protein were composed of 12 subunits and 17 subunits in the muscle of instantaneously killed prawn ana were changed into 8 subunits and 22 subunits in the muscle stored for 26 days, respectively. It is noticeable that 30,000, 41,000, 107,000, 136,000, 170,000 173,000, 185,000, and 198,000 daltons of the newly appeared 8 subunits were found in the myofibrillar protein from the prawn muscle stored for 26 days. The amino acid composition of the muscle protein showed that the most of amino acids were slightly decreased with the days of the storage. With respect to the free amino acid composition of the muscle of instantaneously killed prawn, glycine, proline, arginine, alanine and taurine comprised $93\%$ of the total free amino acids. Taurine, valine, leucine, phenylalanine, serine, lysine, methionine, isoleucine and histidine were increased during the storage period but exceptionally proline was decreased.

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