• 제목/요약/키워드: molecular profile

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비둘기 및 수생조류(水生鳥類) 유래(由來) Salmonella typhimurium의 생물화학적(生物化學的) 특성(特性)과 plasmid profile에 관(關)한 연구(硏究) (Studies on the biochemical characteristics and plasmid profiles of Salmonella typhimurium isolated from pigeons and aquatic birds)

  • 박노찬;최원필
    • 대한수의학회지
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    • 제30권2호
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    • pp.203-214
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    • 1990
  • A total of 166 strains of Salmonella (S) typhimurium var copenhagen isolated from pigeons (164 strains) and aquatic birds (2 strains) were examined for the biochemical characteristics and plasmid profiles. All the strains were sensitive to ampicillin, chloramphenicol, gentamicin, kanamycin and sulfadimethoxine. But 13 strains(7.8%) were resistant to streptomycin (Sm), 2 (1.2%) to tetracycline, 2 (1.2%) to rifampicin, and 1 (0.6%) to nalidixic acid. Among drug resistant strains, only one strain resistant to Sm contained conjugative R plasmid which was fertility inhibition and incompatibility group $I_{\alpha}$. All the strains were sensitive to cobalt chloride, cupric sulfate, lead nitrate, mercuric chloride and silver nitrate. Of 166 isolates, 6 (3.6%) were resistant to sodium arsenate and 1 (0.6%) to potassium tellurite. Among 166 isolates, 1 (0.6%) was colicinogenic, 12 (7.2%) sucrose fermenters, and 166 (100%) maltose fermenters. Plasmid profiles were confirmed as being 4 or 5 plasmids, and their molecular weight ranged 3.2 to 60 megadalton (MD). All the strains harbored 60 Md plasmid. There are three patterns by the plasmid profile, 150 isolates (90.4%) were pattern I (3.2, 3.5, 33, 60Md), 14 (8.4%) pattern II (3.2, 3.5, 29, 60Md), and 2 (1.2%) pattern III (4.2, 7.8, 8.5, 15, 60Md). S typhimurium var copenhagen strains containing 60Md plasmid were resistant to killing by 90% normal guinea pig serum.

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기존의 핵산증폭기를 사용한 유리슬라이드상에서의 원위치 중합효소 연쇄반응 (In Situ PCR on the Glass Slide Using the Conventional DNA Thermal Cycler)

  • 오정균;장진수;이재영
    • 미생물학회지
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    • 제39권3호
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    • pp.197-200
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    • 2003
  • 기존의 핵산증폭기를 사용하여 유리 슬라이드상에서의 효과적인 원위치 중합효소 연쇄반응(in situ PCR)을 수행하기 위해서는 여러 가지 조건들을 고려해야 하는데, 이러한 조건에는 PCR 용액의 세포속으로의 침투, 증폭된 PCR산물의 세포외 유출의 방지, PCR용 액 성분의 유리슬라이드로의 비특이적 부착으로 인한 손실, 열에 의한 시약의 증발, heat block으로부터 슬라이드로의 열전도성 둥이 있다. 특히 PCR용액 성분의 세포내로의 침투를 보장하기 위해서는 다소 높은 농도의 PCR 용액성분(특히 4.5 mM $MgCl_{2}$ 농도)이 필요하였고, Taq 효소는 PCR전 처리(pre-PCR incubation)를 수행하는 경우,50 ${\mu}l$ 반응당 5~10 units이면 충분하였다. 또한 PCR의 전형적인 온도-시간 양상(temperature-time profile)을 만족시키기 위해서는 먼저 샘플의 건조화를 방지해야 하는데, 이를 위해서 heat block속의 빈 공간에 적당량의 물을 첨가했고, 설정온도와 실제온도를 측정해본 결과 약3~$4^{\circ}C$의 차이가 있었다.

Combined Expression of Metastasis Related Markers Naa10p, SNCG and PRL-3 and its Prognostic Value in Breast Cancer Patients

  • Min, Li;Ma, Ruo-Lan;Yuan, Hua;Liu, Cai-Yun;Dong, Bing;Zhang, Cheng;Zeng, Yan;Wang, Li;Guo, Jian-Ping;Qu, Li-Ke;Shou, Cheng-Chao
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권7호
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    • pp.2819-2826
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    • 2015
  • Combinations of multiple biomarkers representing distinct aspects of metastasis may have better prognostic value for breast cancer patients, especially those in late stages. In this study, we evaluated the protein levels of N-${\alpha}$-acetyltransferase 10 protein (Naa10p), synuclein-${\gamma}$ (SNCG), and phosphatase of regenerating liver-3 (PRL-3) in 365 patients with breast cancer by immunohistochemistry. Distinct prognostic subgroups of breast cancer were identified by combination of the three biomarkers. The Naa10p+SNCG-PRL-3-subgroup showed best prognosis with a median distant metastasis-free survival (DMFS) of 140 months, while the Naa10p-SNCG+PRL-3+subgroup had the worst prognosis with a median DMFS of 60.5 months. Multivariate analysis indicated Naa10p, SNCG, PRL-3, and the TNM classification were all independent prognostic factors for both DMFS and overall survival (OS). The three biomarker combination of Naa10p, SNCG and PRL-3 performed better in patients with lymph node metastasis, especially those with more advanced tumors than other subgroups. In conclusion, the combined expression profile of Naa10p, SNCG and PRL-3, alone or in combination with the TNM classification system, may provide a precise estimate of prognosis of breast cancer patients.

Microsatellite 마커를 이용한 딸기 품종의 DNA Profile Database 구축 (Construction of DNA Profile Data Base of Strawberry Cultivars Using Microsatellite Markers)

  • 홍지화;최근진;권용삼
    • 원예과학기술지
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    • 제32권6호
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    • pp.853-863
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    • 2014
  • 국내외에서 재배되고 있는 딸기 100품종의 DNA profile 데이터베이스를 구축하기 위하여 다형성이 높은 microsatellite 마커의 선정과 유전적 유사도 분석을 통한 품종식별력 검정 등에 대한 연구를 수행하였다. 딸기 21품종을 274개의 microsatellite 마커로 검정하여 반복 재현성이 높은 25개의 다형성이 높은 마커를 선정하였다. 이들 마커와 국내외에서 재배되고 있는 딸기 100품종을 검정하였을 때 마커당 평균 대립유전자수는 7.50개로 나타났고, 3-13개까지 다양한 분포를 나타내었다. PIC 값은 마커의 유전자형에 따라 0.333-0.841 범위에 속하였으며 평균값도 0.706으로 높게 나타났다. Microsatellite 마커의 대립유전자를 이용하여 딸기 100 품종에 대한 계통도를 작성하였을 때 품종 육성의 계보 및 육성 지역에 따라 7개의 그룹으로 크게 나누어졌으며 2품종을 제외한 98품종이 microsatellite 마커의 유전자형에 의해 식별이 되는 것으로 나타났다. 본 연구에서 얻어진 딸기 품종별 DNA profile 데이터베이스는 품종보호 출원 품종의 재배심사 및 품종진위성과 관련된 종자분쟁을 해결하는 수단으로 유용하게 활용될 수 있을 것이다.

Molecular Mechanism of Crocin Induced Caspase Mediated MCF-7 Cell Death: In Vivo Toxicity Profiling and Ex Vivo Macrophage Activation

  • Bakshi, Hamid A;Hakkim, Faruck Lukmanul;Sam, Smitha
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권3호
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    • pp.1499-1506
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    • 2016
  • Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

Effects of fermentation on protein profile of coffee by-products and its relationship with internal protein structure measured by vibrational spectroscopy

  • Samadi;Xin Feng;Luciana Prates;Siti Wajizah;Zulfahrizal;Agus Arip Munawar;Peiqiang Yu
    • Animal Bioscience
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    • 제36권8호
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    • pp.1190-1198
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    • 2023
  • Objective: To our knowledge, there are few studies on the correlation between internal structure of fermented products and nutrient delivery from by-products from coffee processing in the ruminant system. The objective of this project was to use advanced mid-infrared vibrational spectroscopic technique (ATR-FT/IR) to reveal interactive correlation between protein internal structure and ruminant-relevant protein and energy metabolic profiles of by-products from coffee processing affected by added-microorganism fermentation duration. Methods: The by-products from coffee processing were fermented using commercial fermentation product, called Saus Burger Pakan, consisting of various microorganisms: cellulolytic, lactic acid, amylolytic, proteolytic, and xylanolytic microbes, for 0, 7, 14, 21, and 28 days. Protein chemical profiles, Cornell Net Carbohydrate and Protein System crude protein and CHO subfractions, and ruminal degradation and intestinal digestion of protein were evaluated. The attenuated total reflectance-Ft/IR (ATR-FTIR) spectroscopy was used to study protein structural features of spectra that were affected by added microorganism fermentation duration. The molecular spectral analyses were carried using OMNIC software. Molecular spectral analysis parameters in fermented and non-fermented by-products from coffee processing included: Amide I area (AIA), Amide II (AIIA) area, Amide I heigh (AIH), Amide II height (AIIH), α-helix height (αH), β-sheet height (βH), AIA to AIIA ratio, AIH to AIIH ratio, and αH to βH ratio. The relationship between protein structure spectral profiles of by-products from coffee processing and protein related metabolic features in ruminant were also investigated. Results: Fermentation decreased rumen degradable protein and increased rumen undegradable protein of by-products from coffee processing (p<0.05), indicating more protein entering from rumen to the small intestine for animal use. The fermentation duration significantly impacted (p<0.05) protein structure spectral features. Fermentation tended to increase (p<0.10) AIA and AIH as well as β-sheet height which all are significantly related to the protein level. Conclusion: Protein structure spectral profiles of by-product form coffee processing could be utilized as potential evaluators to estimate protein related chemical profile and protein metabolic characteristics in ruminant system.

Computational Study on Oligomer Formation of Fibril-forming Peptide of α-Synuclein

  • Park, Seong-Byeong;Yoon, Je-Seong;Jang, Soon-Min;Lee, Kyung-Hee;Shin, Seok-Min
    • Bulletin of the Korean Chemical Society
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    • 제33권3호
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    • pp.848-854
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    • 2012
  • We have studied the oligomerization of a fibril-forming segment of ${\alpha}$-Synulcein using a replica exchange molecular dynamics (REMD) simulation. The simulation was performed with trimers and tetramers of a 12 amino acid residue stretch (residues 71-82) of ${\alpha}$-Synulcein. From extensive REMD simulations, we observed the spontaneous formation of both trimer and tetramer, demonstrating the self-aggregating and fibril-forming properties of the peptides. Secondary structure profile and clustering analysis illustrated that antiparallel ${\beta}$-sheet structures are major species corresponding to the global free energy minimum. As the size of the oligomer increases from a dimer to a tetramer, conformational stability is increased. We examined the evolution of simple order parameters and their free energy profiles to identify the process of aggregation. It was found that the degree of aggregation increased as time passed. Tetramer formation was slower than trimer formation and a transition in order parameters was observed, indicating the full development of tetramer conformation which is more stable than that of the trimer. The shape of free energy surface and change of order parameter distributions indicate that the oligomer formation follows a dock-and-lock process.

다상 유체 시스템의 다중 스케일 시뮬레이션 기법에 관한 연구 (Development of Multiscale Simulation Technique for Multiphase Fluid System)

  • 한민섭
    • 대한기계학회논문집B
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    • 제34권6호
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    • pp.569-577
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    • 2010
  • 다상 유체 시스템에 적용할 수 있는 다중스케일 입자 시뮬레이션 기법을 개발하였다. 거시 모델과 미시모델이 만나는 경계영역에서 세가지로 구별되는 기능을 수행하도록 하였다. 먼저, 기상과 액상을 분리하여 연결하기 위해 벽을 설정하였다. 또 경계영역을 근처에서 경계의 위치를 측정하고 이것에 벽의 각도와 위치가 연동하여 접촉각 값을 미시모델에서 거시모델로 전달하게 하였다. 마지막으로, 입자의 삽입과 제거를 통해 경계영역의 질량과 온도를 거시적 조건에 맞추도록 하였다. 이 알고리즘들을 적용한 완전습윤과 부분습윤시스템들은 좋은 결합 결과를 보였다.

아스팔트 시멘트의 분자립도 분포특성에 따른 아스팔트의 성질분석 (Estimation of Bituminous Concrete Property Based on Molecular Size Distribution of Asphalt Cement)

  • 김광우;연규석
    • 콘크리트학회지
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    • 제3권1호
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    • pp.71-78
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    • 1991
  • 3종류의 AC-20 아스팔트 시멘트의 분자입자분포를 고압-겔투수 크로마토그래프(HP-GPC)를 이용하여 관찰하였다. 건조상태의 습윤상태의 인장강도와 뤠질리언계수 측정을 위하여 한종류의 골재와 아스팔트 시멘크를 사용하여 실험실에서 역청콘크리트 공시체를 제조하였다. 이들의 역학적 성질시험결과와 아스팔트 시멘트 크로마토그램의 형태를 분석하여 상호간의 관계를 조사하였다. 희귀분석 결과는 역학적 성질과 크로마토그램의 분할면적비율간에 강한 상관관계가 있음을 보여주었다. 이 희귀분석 결과로부터 수학적 추정모델이 얻어졌다. 연구결과 아스팔트 크로마토그램의 특성이 정해진 아스팔트콘크리트의 역학적 성질추정이 잘 사용될 수 있음을 보여주었다.

Toxicoproteomic Analysis of Differentially Expressed Proteins in Rat Liver by DEHP

  • Son, Bu-Soon;Seong, Ah-Reum;Park, Seul-Ki;Kim, Wan-Jong;Ryu, Jae-Chun;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • 제3권4호
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    • pp.299-305
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    • 2007
  • The endocrine disrupting chemical, di (2-ethylhexyl) phthalate (DEHP) is a plasticizer used in polyvinyl chloride products ubiquitous in our daily lives. DEHP has potentially adverse effects on the liver, kidney, lung, heart, reproductive organs and endocrine systems. Many toxicological data on the DEHP toxicity have been stated, but complete protein profiles have not yet been reported. In this study, DEHP-induced oxidative DNA damage in rat lymphocyte was evaluated by Comet assay (single-cell gel electrophoresis) for the first time. Moreover, DEHP-induced protein profile alterations were examined in rat liver by using toxicoproteomic tools. 34 protein spots in the liver were identified to be significantly deregulated by DEHP on the 2-dimensional gel. Among them, 20 spots were up-regulated and 14 spots down-regulated by DEHP.