• Title/Summary/Keyword: e-spots

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이차원전기영동을 이용한 Escherichia coli O157:H7 균주간 항원 Spot의 비교 (Comparison of Antigenic Spots between Escherichia coli O157:H7 Strains by 2-Dimensional Gel Electrophoresis)

  • 안영창;신기욱;신용승;이응구;이형준;박미림;김영림;정태성;김곤섭;김용환
    • 대한수의학회지
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    • 제42권2호
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    • pp.231-239
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    • 2002
  • Proteomics is an emerging powerful tool in studying protein expression and function. At present study, proteomics was employed to evaluate the antigenicity among Escherichia coli O157:H7 strains using 2-dimensional gel electrophoresis (2-DE) and immunoblotting, SDS-PAGE and immunblotting analysis revealed no big differences among E coli O157:H7 strains. 2-DE analysis, however, revealed common antigens as well as specific antigens. The immunoblotting analysis revealed 20 common antigenic spots among E coli O157:H7 strains. In addition, there were 3 and 13 spots as common antigens between ATCC 43894 and KSC 109, and between ATCC 43894 and ACH 5, respectively. Antigenic spots specific for individual strain were also identified as 15, 8 and 22 for ATCC 43894, ACH 5 and KSC 109, respectively. The common antigens would be useful by employing either vaccine development or diagnosis marker, or both, whereas the specific antigens of individual strains would be applicable for epidemiological study. This study suggest that proteome analysis, representative as 2-DE, is valuable tool in exploring the E. coli antigenicity.

외대버섯속의 담자포자의 발생과 미세구조 (Basidiospore Development and Fine Structure of Entoloma)

  • 조덕현
    • 한국자원식물학회지
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    • 제6권2호
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    • pp.141-145
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    • 1993
  • Apexes of basidia in Rhodophyllus(synonyum of Entoloma) muraii var. albus are divided into four sections or depressed in the center. A spore is formed by inflation of the apex of the sterigma. The apex of the sterigma is swollen and changed from a papilla through a penisform and a club into a globeform. Six spots of globose spore are regularly or irregularly depressed with hilum axes. Finally the spores are cuboid. Basidia of Entoloma squamiferrum are developed from hymenium layer of crator-shaped parabasidium. Apex of basidium is flat or depressed in the center. Although four sterigmata are developed, only two sterigmata are symmetrically swollen to two spores in certain basidia. It means that two sterigmata among four sterigmata are infertile. The apex of the sterigma is swollen into a paillaform, and then turned into a penisform. It is swollen from a clubform into a globeform. Six spots of the surfaces of globose spore are deperssed with hilum axes. Finally the spore is cuboid, and then it is released from hilum. Four sterigmata of papillaform of E. violaceobrunneum are developed from cartor-shaped basidium. The apex of sterigma is swollen to a small globeform. And then it is swollen to a clubform. The clubform is again swollen to a ellipticalform, and then more than six spots of spore surfaces are ramomly depressed with hium axes. When the depression of surface of a elliptical spore is over, it is the multi-angular spore of the heterometrical-form.

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Proteomic Analysis in ob/ob Mice Before and After Hypoglycemic Polysaccharide Treatments

  • Kim, Sang-Woo;Hwang, Hye-Jin;Baek, Yu-Mi;Hwang, Hee-Sun;Yun, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • 제19권10호
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    • pp.1109-1121
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    • 2009
  • In an attempt to discover novel biomarker proteins in type 2 diabetes prognosis, we investigated the influence of hypoglycemic extracellular polysaccharides (EPS) obtained from the macrofungus Tremella fuciformis on the differential levels of plasma proteins in ob/ob mice using two-dimensional gel electrophoresis (2-DE). The 2-DE analysis demonstrated that 92 spots from about 900 visualized spots were differentially regulated, of which 40 spots were identified as principal diabetes-associated proteins. By comparing control with EPS-fed mice, we found that at least six proteins were significantly altered in ob/ob mice, including Apo A-I, IV, C-III, E, retinol-binding protein 4, and transferrin, and their levels were interestingly normalized after EPS treatment. Western blot analysis revealed that the altered levels of the two regulatory molecules highlighted in diabetes and obesity (e.g., resistin and adiponectin) were also normalized in response to EPS. The Mouse Diabetes PCR Array profiles showed that the expression of 84 genes related to the onset, development, and progression of diabetes were significantly downregulated in liver, adipocyte, and muscle of ob/ob mice. EPS might act as a potent regulator of gene expression for a wide variety of genes in ob/ob mice, particularly in obesity, insulin resistance, and complications from diabetes mellitus.

Immunochemical Studies on Expression of Quinoproteins in Escherichia coli

  • Ryou, Chong-Suk;Kim, Jae-Beom;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • 제10권1호
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    • pp.95-98
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    • 2000
  • An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

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Coupled temperature-displacement modeling to study the thermo-elastic instability in disc brakes

  • Ramkumar, E.;Mayuram, M.M.
    • Coupled systems mechanics
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    • 제1권2호
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    • pp.165-182
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    • 2012
  • Macroscopic hot spots formed due to the large thermal gradients at the surface of the disc brake rotor, make the rotor to fail or wear out early. Thermo-elastic deformation results in contact concentration, leading to the non uniform distribution of temperature making the disc susceptible to hot spot formation. The formation of one hot spot event will predispose the system to future hot spotting at the same location. This leads to the complete thermo-elastic instability in the disc brakes; multitude parameters are responsible for the thermo elastic instability. The predominant factor is the sliding velocity and above a certain sliding velocity the instability of the brake system occurs and hot spots is formed in the surface of the disc brake. Commercial finite element package ABAQUS(R) is used to find the temperature distribution and the result is validated using Rowson's analytical model. A coupled analysis methodology is evolved for the automotive disc brake from the transient thermo-elastic contact analysis. Temperature variation is studied under different sliding speeds within the operation range.

유충의 생식후원선이 아닌 한 쌍의 소반점에 관한 조사연구 (1) (Study on a pair of small spot of nonsexual gland hind of the larvae in Bombyx mori L. (I))

  • 윤종관
    • 한국잠사곤충학회지
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    • 11호
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    • pp.51-54
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    • 1970
  • 유충기와 용기는 성충기에 비하여 외 상대징이 획연치 않기 때문에 유충기에 있어서의 현행 자웅감별 방법은 5령의 2∼3일째에 비교적 뚜렷이 관찰 할 수 있는 수컷의 Herold's gland와 암컷의 석도생식원선 (생식전선과 생식후선)에 의하여 이루어 지고 있다. 그러나 품종에 따라서는 수컷의 제9복절의 복면에 한쌍의 소반점이 있고 이 소반점의 위치가 암컷에서 볼 수 있는 생식후원선의 위치와 일치하기 때문에 암컷으로 오인되기 쉬워 자웅감별에 있어 혼란을 이르키고 있으므로 이를 조사연구 하였는데 이를 요약하면 다음과 같다. 1. 수컷의 제9복절에 있는 소반점의 유무는 품종에 따라 다르다. 계통별로 보아 중국종은 일본종에 비하여 소반점이 나타나는 개체가 많을뿐더러 전감별 수컷의 과반수를 초과한 57.0%를 차지 하고 있다. 2. 소반점의 형상은 암컷의 생식후원선과 같이 대개 원형을 이루고 있지만 빛깔에 있어서는 생식후원선은 불투명한 유백색이며 소반점은 품종에 따라 다소 다르다고는 하나 약간의 회백색을 띄우고 있다. 그리고 이 소반점은 생식후원선에 비하기 소형이다. 누에의 생식선은 숙잠기에 이르면 식별할 수 없게 되지만 이 소반점은 숙잠기에 이르러도 육면으로 구별할 수 있고 또 생식후원선의 위치에 있다. 3. 소반점과 Herold's gland를 기초로 하여 감별을 한 다음 그 정확성을 다시 확인하기 위하여 화용후에 개체감별을 한 결과를 보면 평균 5.2%의 이성이 혼입되어 있다. 이러한 까닭은 소반점의 위치가 암컷에 있는 생식후원선의 위치와 일치하기 때문인 것으로 생각된다. 4. 소반점의 유무와 농담 등에 의하여 화용 후에 있어서의 활동상황 및 생식능력에 있어서는 하등의 이상이 없었다. 5. 소반점의 유무는 계통별로 차이가 있을뿐더러 품종간의 교잡시에 유전적인 연관성도 있을 것이고 화성 및 면성 등에 의해서도 그 출현상태지 차이가 있을 것으로 추정된다.

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Nano-scale Proteomics Approach Using Two-dimensional Fibrin Zymography Combined with Fluorescent SYPRO Ruby Dye

  • Choi, Nack-Shick;Yoo, Ki-Hyun;Yoon, Kab-Seog;Maeng, Pil-Jae;Kim, Seung-Ho
    • BMB Reports
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    • 제37권3호
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    • pp.298-303
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    • 2004
  • In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).

Erwinia chrysanthemi에 의한 Ivy-aureus (Scindapsus aureus)의 세균성 갈색부패병 (Bacterial Brwon Rot of Ivy-aureus (Scindapsus aureus) Caused by Erwinia chrysanthemi)

  • 최재을;한광섭
    • 한국식물병리학회지
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    • 제10권4호
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    • pp.336-338
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    • 1994
  • In 1991, the leaf brown spot of ivy-aureus (Scindapsus aureus) was found in Taejon and Seoul, Korea. The symptoms were appeared as dark-brown spots. The lesions were often surrounded by yellowish halos. These spots were enlarged to circular or elliptical in shape and dark-brown to black in color with slightly elevated in margin and sunken in center. The pathogenic bacteria were isolated from the diseased leaf of ivy-aureus were identified as Erwinia chrysanthemi on the basis of bacterial characteristics therefore, we would like to propose to the name of ivy-aureus disease caused by E. chrysanthemi as“bacterial brown rot of ivy-aureus”hereafter.

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회전형 저온 플라즈마 시스템을 이용한 분말식품의 균일한 살균 연구 (A Study of Homogeneous Sterilization of Micro-sized Food Powder by Rotatable Low-Temperature Plasma System)

  • 김명찬;박덕모;한진수;우인봉;김동후;장성은;윤찬석;김인
    • 한국식품영양학회지
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    • 제31권2호
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    • pp.301-307
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    • 2018
  • In this study, a relatively effective process is used to sterilize Escherichia coli on the surface of micro-sized calcium citrate powder using nitrogen and argon as process gases in a low-temperature vacuum plasma treatment. The purpose of this study is to confirm and to introduce the effectiveness of homogeneous surface treatment for the sterilization of fine inorganic powder by the rotatable low-temperature RF plasma system designed by ourselves. The results of the test using 3M petrifilm showed that there were no remarkable spots in the case of the surface of plasma treated powder, whereas the untreated powder showed many blue spots, which indicating that the E. coli was alive. After 5 days, in the same samples, the blue spots were seen to be larger and darker than before, while the plasma-treated powder showed no changes. The results from FE-SEM analysis showed that the E. coli was damaged and/or destroyed by reactive species generated in the plasma space, resulting in the E. coli being sterilized. Furthermore, the sterilization effects according to the selected parameters ($N_2$ and Ar; flow rate 30 and 50 sccm) adapted in this study were mutually similar, regardless of such different process parameters, and this indicates that homogeneous treatment of powder surfaces could be more effective than conventional methods. Therefore, the plasma apparatus used in this study may be a practical method to use in a powerful sterilization process in powder-type food.

Profiling of differential expressed proteins from various explants in Platycodon grandiflorum

  • Kim, Hye-Rim;Kwon, Soo Jeong;Roy, Swapan Kumar;Kamal, Abu Hena Mostafa;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Cho, Kab Yeon;Woo, Sun-Hee
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.131-131
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    • 2017
  • Though the Platycodon grandiflorum, has a broad range of pharmacologic properties, but the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two-dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}2-fold$) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, the frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). Taken together, the protein profile may provide insight clues for better understanding the characteristics of proteins and its metabolic activities in various explants of this essential medicinal plant P. grandiflorum.

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