• Title/Summary/Keyword: gel image analysis

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Proteomic Analysis of MCAo Ischemia Model Administered with Yukmijihwangtang (MCAo 허혈동물모델에서 육미지황탕 효능에 관한 프로테오믹스 연구)

  • Kim, Young-Ok;Cho, Dong-Wuk;Kang, Bong-Joo
    • Korean Journal of Oriental Medicine
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    • v.13 no.1 s.19
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    • pp.153-160
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    • 2007
  • In the post-genome era, analysis of the cellular transcriptome using microarray or the cellular proteome using a 2-D gel electrophoresis and MALDI-TOF mass spectrometry are most widely used. Stroke is one of the most important causes of death along with cancer and cardiac disease. When pathological change of cells in developed from cerebral ischemia accompanied by stroke administration of neuroprotective drugs before stroke can decreases the degeneration of neuronal cells. The purpose of the present study was to assess the neuroprotective effect and protein expression after administration of P004, middle cerebral artery model of cerebral ischemia in rats. SD rats were subjected to middle cerebral artery occlusion. P004 (1,000 mg/kg) was administered 2 times at 0, 90 minutes after middle cerebral artery occlusion (MCAo). Rats were killed at 48 hours, and infarct area and volume were determined by histology and computerized image analysis. We investigated the protein expression profile on the global ischemia induced by MCAo. This proteomic analysis enable us to identify several proteins differently expressed in infarct brain tissue. The aims of this study were to do investigation comparing the neuroprotection activities of P004 and to understand the mechanism of acted as neuroprotective drug.

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Catalytic combustion of $H_2$/Air mixture using Pt/$Al_2O_3$ coated nickel foam (Pt/$Al_2O_3$가 코팅된 니켈폼을 이용한 수소-공기 예혼합 기체의 촉매 연소)

  • Jin, Jung-Kun;Kwon, Se-Jin
    • 한국연소학회:학술대회논문집
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    • 2007.05a
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    • pp.37-44
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    • 2007
  • A nickel foam, one of metal foams was seleced as a catalyst support instead of conventional ceramic materials. $Al_2O_3$ was coated on the surface of nickel foam to increase the surface area. $Al_2O_3$ coating process was based on sol-gel process. SEM image was obtained and $Al_2O_3$ coverage was confirmed. Combustion experiments were carried out using SUS combustor and $H_2$/air mixture. Temperatures were measured with different equivalence ratios and $H_2$ flow rates. $H_2$ conversion rates were calculated by the analysis of product gas using gas chromatography. Catalytic combustion of $H_2$ was complete and stable with Pt/$Al_2O_3$ coated nickel foam and influences of water vapor were confirmed during the beginning of combustion.

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Data Analysis Methods for Quantitative Proteomics Research

  • Gwon Kyeong-Hun
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2006.02a
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    • pp.38-44
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    • 2006
  • 프로테오믹스는 생물체 안에 포함되어 있는 단백질을 통합적으로 연구하는 학문이다. 단백질을 동정(Protein identification)하고, 단백질의 상태를 분석(Protein characterization)하며, 단백질의 양적 변화를 관찰(Protein quantitation)한다. 유전자로부터 mRNA 로 복제되고 codon 의 규칙에 따라 합성되는 단백질이 세포 내에 얼만큼 존재하는가라는 단백질의 양적인 변화는 세포 내의 환경에 따라 시시각각 변화할 수 있으며, 이러한 변화의 추적은 단백질의 기능을 밝히는 기초자료로서 중요성을 가진다. 특히 질병의 조기 진단을 위한 바이오마커를 발굴하기 위한 스크리닝 역할로서, 단백질의 발현 양상을 비교하는 프로테오믹스는 기대를 모으고 있다. 단백질에 대한 분석, 특히 질량분석기에 의해 초고속으로 대량의 단백질 데이터를 생산하는 프로테오믹스의 연구는 정량적인 단백질 발현양상 분석의 정확도를 높이기 위해 다양한 실험기법과 데이터 분석기법을 동원하고 있다. 이번 발표에서는 프로테오믹스에서 단백질의 양을 측정하기 위한 실험 방법들과 그에 따른 데이터 분석 방법들을 소개하고자 한다. 프로테오믹스 연구의 초창기부터 사용되어온 2차원 전기영동법에 의해 생성되는 2D-gel image 에서의 spot 분석법으로부터, 탄뎀 질량분석기를 사용하는 ICAT, iTRAQ 등의 labeling 방법에 의한 정량분석, 그리고 질량분석기의 정확도를 최대한으로 활용하는 label-free 방법에 대한 기본 개념을 살펴보고 데이터 분석 기술의 적용 방법을 알아본다.

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Microstructural Study of Mortar Bar on Akali-Silica Reaction by Means of SEM and EPMA Analysis (알칼리-실리카 반응에 의한 모르타르 봉의 SEM과 EPMA 분석을 통한 미세구조 연구)

  • Jun, Ssang-Sun;Lee, Hyo-Min;Jin, Chi-Sub
    • Journal of the Korea Concrete Institute
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    • v.21 no.4
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    • pp.531-537
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    • 2009
  • In this study alkali reactivity of crushed stone was conducted according to the ASTM C 227 that is traditional mortar bar test, and C 1260 that is accelerated mortar bar test method. The morphology and chemical composition of products formed in mortar bar, 3 years after the mortar bar tests had been performed, were examined using scanning electron microscopy (SEM) with secondary electron imaging (SEI) and electron probe microanalysis (EPMA) with backscattered electron imaging (BSEI). The crushed stone used in this study was not identified as being reactive by ASTM C 227. However, mortar bars exceeded the limit for deleterious expansion in accelerated mortar bar test used KOH solution. The result of SEM (SEI) analysis, after the ASTM C 227 mortar bar test, confirmed that there were no reactive products and evidence of reaction between aggregate particles and cement paste. However, mortar bars exposed to alkali solution (KOH) indicated that crystallized products having rosette morphology were observed in the interior wall of pores. EPMA results of mortar bar by ASTM C 227 indicated that white dots were observed on the surface of particles and these products were identified as Al-ASR gels. It can be considered that the mortar bar by ASTM C 227 started to appear sign of alkali-silica reaction in normal condition. EPMA results of the mortar bar by ASTM C 1260 showed the gel accumulated in the pores and diffused in to the cement matrix through cracks, and gel in the pores were found to be richer in calcium compared to gel in cracks within aggregate particles. In this experimental study, damages to mortar bars due to alkali-silica reaction (ASR) were observed. Due to the increasing needs of crushed stones, it is considered that specifications and guidelines to prevent ASR in new concrete should be developed.

Comparison of Specific Proteins of Shiga Toxin-producing E. coli (STEC) Adhesion by Lactobacillus acidophilus Strains Using Two Dimensional Gel Electrophoresis (이차원 전기영동을 이용한 Lactobacillus acidophilus Strains의 Shiga Toxin-producing E. coli (STEC) 부착 억제와 관련된 단백질 발현 변화 분석)

  • Kim Young-Hoon;Moon Yong-Il
    • Food Science of Animal Resources
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    • v.26 no.2
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    • pp.263-268
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    • 2006
  • Probiotics including Lactobacillus acidophilus, refer to a group of nonpathogenic organisms that protect the human host against gastrointestinal(GI) infections by pathogenic bacteria such as Shiga toxin-producing E. coli(STEC). In the study, the inhibitory effects of STEC ATCC 43894 adhesion by L. acidophilus A4 was investigated on the HT-29 epithelial cells. Specific proteins regulated by cell Iysates of L. acidophilus A4 on STEC ATCC 43894 were also characterized by proteomic analysis. Both cell mass and Iysate of L. acidophilus A4 have exhibited the profound inhibitory activity on the HT-29 cells(about 1.5 log scale reduction). Two-dimensional gel electrophoresis(2-DE) revealed seven proteins that were up-regulated by cell Iysates of L. acidophilus A4 and three proteins that were down-regulated. In addition, three protein spots were only detected in the presence of cell Iysates. These results suggest that inhibitory effects of STEC adhesion by L. acidophilus may be due to the regulation of specific protein of STEC.

Purification and Structural Characterization of P93 Complex from Hyperthermophilic Archaeon Thermococcus profundus (초고온 archaeon인 Thermococcus profundus에서 P93 복합체의 분리 및 구조적 특성)

  • Lee, Mi-Hong;Kim, Suk-Kyoung;Yun, Young-Gun;Park, Seong-Cheol;Bahk, Jeong-Dong;Cheong, Gang-Won
    • Applied Microscopy
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    • v.30 no.2
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    • pp.185-191
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    • 2000
  • An unusually large protein complex was found in the cytosol of the hyperthmophilic archaeon. Thermococcus profundus. The purified protein was shown to be a homomultimer of 93 kDa subunit (P93 complex). The complex is extremely heat stable. During 12 hrs incubation with SDS (final concentration 1%) at $85^{\circ}C$, no changed structure could be observed. Electron image analysis of negatively stained showed that the complex has a single, stable characteristic view and a well-preserved core with threefold rotational symmetry. The periphery of the assembly is composed of a nebulose, possibly flexible, component. Based on the projected structure suggest the P93 complex from T. profundus is composed 24 homomultimer.

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Isolation and Characterization of the Streptococcus mutans from Korean Children with Caries

  • Kang, Kyung-Hee;Kim, Ji-Young;Nam, Jin-Sik;Jin, Ingn-Yol
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.341-347
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    • 2007
  • Streptococcus mutans, one of a major causal agents of dental caries, is component of the dental plaque and produces various organic acids such as lactic acid as the end-product of glycolysis. In this study, we isolated S. mutans from Korean children with caries and also investigated the expression of protein under acid stress. S. mutans was identified at the species level using a 16S ribosomal DNA sequencing comparison method. The primer specificity was tested on eleven S. mutans strains isolated from Korean children with caries. The data showed that eleven strains are S. mutans. At treatment of concentration of 20 mM lactic acid in the mid-log phage, K-7 exhibited the highest maximum culture OD compared with those of other groups. As a consequence, we examined the expression of protein under 20 mM lactic acid stress using S. mutans K-7. The results of 2D gel electrophoresis by image analysis showed that thirteen proteins are up-regulated under the stress. Further study is being focused on amino acid analysis by mass spectrometry in order to analyze those spots.

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Mechanism study on DNA damage and Apoptosis induced by heak shock using Comet Assay

  • Seo, Young-Rok;Han, Sung-Sik;Kim, L. O′Neill;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 1997.12a
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    • pp.101-101
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    • 1997
  • Comet assay, single cell gel electrophoresis has been known as useful, rapid, simple, visual, and sensitive technique for measuring the DNA breakage in mammalian ce1ls. For evaluation of DNA damage using comet assay, early studies reported a change in comet length and intensity with DNA damage using simple visual technique, such as fluorescence microscopy with eyespiece. In recent, some workers are observing and analyzing nucleotide of comets using quantitative fluorescence image analysis system to estimate 'tail moment', which is defined as the product of the tail length and the fraction of total DNA in tail. Our laboratory also adopted the image analysis software for qualification. In addition, many of the practical features of comet assay render it potentially attractive as useful tool for molecular toxicology and carcinogenesis, because the system is already showing considerable promise as rapid predictor in both in vitro and in vivo experimental designs. Recently, the comet assay becomes a attractive technique to study of apoptosis, because apoptotic fragmentation of nuclear DNA into nucleosomal sizes can be evaluated by the comet assay. So, we attempted to apply the comet assay to studying the effect of various stress on the apoptosis-sensitive cell lines. Particularly, focusing on the hyperthermic apoptosis, we could find that heat shock(44˚C for 60 minutes) was sufficient to induced apoptosis in these cell lines. But using the highly sensitive comet assay, we could not detect DNA breaks immediately after heat shock.

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Expression of AGR-2 in Chicken Oviduct during Laying Period

  • Kim, Nam-Soo;Shen, Yan-Nan;Kim, Tae-Yoon;Byun, Sung-June;Jeon, Ik-Soo;Kim, Sang-Hoon
    • BMB Reports
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    • v.40 no.2
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    • pp.212-217
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    • 2007
  • The chicken oviduct is a dynamic organ that produces secretory proteins such as ovalbumin during the laying period. In this study, we identified oviduct-specific proteins in hens during the egg-laying period by proteomic analysis. Proteins extracted from the magnum of hens of different ages (5, 35, and 65 weeks) were analyzed by two-dimensional gel electrophoresis to compare the intensity of proteins among samples. Approximately 300 spots were detected on each gel. Based on the comparison of image gels, we found that the intensity of eight spots in 35-week magnums was increased at least by 2-fold compared with the others. Five of the eight spots were identified as calumenin, acidic ribosomal phosphoproteins (ARP), prohibitin, heart fatty acid-binding protein, and anterior gradient-2 (AGR-2). In particular, ARP and AGR-2 were highly expressed in 35- week magnums compared with 5- and 65-week magnums. In addition, the level of these proteins was consistent with their RNA levels. Expression of AGR-2 mRNA was detected in the mature magnum, whereas no signal was observed in premature tissue. Among various tissues, expression of AGR-2 mRNA was highest in the magnum, high in the isthmus, and five fold lower in muscle. It was undetectable in the liver and in other tissues (heart and kidney). However, the mRNA levels of other proteins were ubiquitous among tissues. In transcriptional activity of AGR-2, a 3.0 kb fragment of promoter region containing potential estrogen receptor binding sites had enhanced its activity strongly. In conclusion, these results suggest that AGR-2 has functional regulatory roles in the chicken oviduct during the egglaying period.

Protein Expression of Mouse Uterus in Post-Implantation

  • Kim, Hong-Rye;Han, Rong-Xun;Kim, Myung-Youn;Diao, Yunfei;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.237-242
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    • 2009
  • Pregnancy is a unique event in which a fetus develops in the uterus despite being genetically and immunologically different from the mother, and the underlying mechanisms remain poorly understood. To analyze the differential gene expression profiles in nonpregnant and 7 days post coitus (dpc) pregnant uterus of mice, we performed a global proteomic study by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. The uterine proteins were separated using 2-DE, Approximately 1,000 spots were detected on staining with Coomassie brilliant blue. An image analysis using Melanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between pregnant and nonpregnant uterus. Twenty-one spots were identified as differentially expressed proteins, of which 10 were up-regulated proteins such as alpha-fetoprotein, chloride intracellular channel 1, transgelin, heat-shock protein beta-1, and carbonic anhydrase II, while 11 were down-regulated proteins such as X-box binding protein, glutathione S-transferase omega 1, olfactory receptor Olfr204, and metalloproteinase-disintegrin domain containing protein TECADAM. Most of the identified proteins appeared to be related with catabolism, cell growth, metabolism, regulation, cell protection, protein repair, or protection. Our results uncovered key proteins of mouse uterus involved in pregnancy.