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Comparative antigen analysis of yrichomonus vaginulis by enzyme-linked immunoelectrotransfer blot technique (효소면역 전기영동 이적법을 이용한 질트리코모나스 항원의 비교 분석)

  • 민득영;임미혜
    • Parasites, Hosts and Diseases
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    • v.30 no.4
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    • pp.323-328
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    • 1992
  • Analysis of siR isolates of Trichemenes veginalis was carried out with sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunoelectrotransfer blot (EITB). Trichloroacetic acid-treated antigens of the 6 isolates revealed 25 protein profiles ranging 12~170 kDa of molecular weight in SDS-PAGE. In EITB, the specific immunogenic bands were visualized at 51 kDa and 96 kDa when HY-1 antigen was probed with difFerent mice sera immunized with 6 isolates of T. vaginalis. The banding patterns with different sera showed isolate-to-isolate variability. In EITB, homologous antigen (HY-1) did not show any enhanced response in reacting to homologous antiserum(HY-1) when 6 isolates of T. vaginalis were probed with a single serum (HY-1). It is assumed that the different banding patterns of six isolates show isolate-to-isolate variability and immunogenic common bands in 41, 47, 74 and 9:k kDa on EITB may connote the important significance on immune response in T. vaginalis infection.

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Diversity of Bacillus thuringiensis Strains Isolated from Citrus Orchards in Spain and Evaluation of Their Insecticidal Activity Against Ceratitis capitata

  • J.C., Vidal-Quist;Castanera, P.;Gonzalez-Cabrera, J.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.749-759
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    • 2009
  • A survey of Bacillus thuringiensis (Berliner) strains isolated from Spanish citrus orchards has been performed, and the strains were tested for insecticidal activity against the Mediterranean fruit fly Ceratitis capitata (Wiedemann), a key citrus pest in Spain. From a total of 150 environmental samples, 376 isolates were selected, recording a total B. thuringiensis index of 0.52. The collection was characterized by means of phase-contrast microscopy, SDS-PAGE, and PCR analysis with primer pairs detecting toxin genes cry1, cry2, cry3, cry4, cry5, cry7, cry8, cry9, cry10, cry11, cry12, cry14, cry17, cry19, cry21, cry27, cry39, cry44, cyt1, and cyt2. Diverse crystal inclusion morphologies were identified: bipyramidal (45%), round (40%), adhered to the spore (7%), small (5%), and irregular (3%). SDS-PAGE of spore-crystal preparations revealed 39 different electrophoresis patterns. All primer pairs used in PCR tests gave positive amplifications in strains of our collection, except for primers for detection of cry3, cry19, cry39, or cry44 genes. Strains containing cry1, cry2, cry4, and cry27 genes were the most abundant (48.7%, 46%, 11.2%, and 8.2% of the strains, respectively). Ten different genetic profiles were found, although a total of 109 strains did not amplify with the set of primers used. Screening for toxicity against C. capitata adults was performed using both spore-crystal and soluble fractions. Mortality levels were less than 30%. We have developed a large and diverse B. thuringiensis strain collection with huge potential to control several agricultural pests; however, further research is needed to find out Bt strains active against C. capitata.

Antigenicity of Protein Entrapped in Poly(lactide-co-glycolide) Microspheres (폴리락티드-글리콜리드 마이크로스피어에 봉입된 단백질의 항원성 평가)

  • Song, Seh-Hyon;Cho, Seong-Wan;Shin, Taek-Hwan;Yoon, Mi-Kyoung;Choi, Young-Wook
    • Journal of Pharmaceutical Investigation
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    • v.31 no.3
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    • pp.191-196
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    • 2001
  • Biodegradable polymeric microspheres were studied for their usefulness as carriers for the delivery of vaccine antigens. However, protein antigen could be denatured during microencapsulation processes due to the exposure to the organic phase and stress condition of cavitation and shear force. Therefore this study was carried out to re-evaluate the degree of protein denaturation during microencapsulation with poly(lactide-co-glycolide) (PLGA) copolymer. PLGA microspheres containing ovalbumin (OVA), prepared by W/O/W multiple emulsification method, were suspended in pH 7.4 PBS and incubated with shaking at $37.5^{\circ}C$. Drug released medium was collected periodically and analyzed for protein contents by micro-BCA protein assay. In order to evaluate the protein integrity, release medium was subjected to the analyses of SDS-PAGE and size exclusion chromatography (SEC). And enzyme-linked immunosorbent assay (ELISA) was introduced to measure the immunoreactivity of entrapped OVA and to get an insight into the three-dimensional structure of epitope. The structures of entrapped protein were not affected significantly by the results of SDS-PAGE and SEC. However, immunoreactivity of released antigen was varied, revealing the possibility of protein denaturation in some microspheres when it was evaluate by ELISA method. Therefore, in order to express the degree of protein denaturation, antigenicity ratio (AR) was obtained as follows: amount of immunoreactivity of OVA/total amount of OVA released ${\times}100(%)$. ELISA method was an efficient tool to detect a protein denaturation during microencapsulation and the comparison of AR values resulted in more accurate evaluation for immunoreactivity of entrapped protein.

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Identification of Different Species and Dultivars of Brassica by SDS-PAGE, Isozyme and Molecular Marker

  • Mukhlesur Rahman Md.;Hirata Yutaka
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.27-35
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    • 2005
  • Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.

[${^3H}Ryanodine$ Binding Sites of SR Vesicles of the Chicken Pectoral Muscle

  • Yun, Hyo-Yung;Jeon, Jong-Rye;Hong, Jang-Hee;Hur, Gang-Min;Lee, Jae-Heun;Seok, Jeong-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.377-384
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    • 1997
  • To investigate the properties of ryanodine binding sites of the bird skeletal SR vesicles, SDS PAGE, purification of RyR, and $[^3H]ryanodine$ binding study were carried out in the SR vesicles prepared from the chicken pectoral muscle. The chicken SR vesicles have two high molecular weight (HMW) protein bands as in eel SR vesicles on SDS PAGE. The HMW bands on SDS PAGE were found in the $[^3H]ryanodine$ peak fraction $(Fr_{3-5})$ obtained from the purification step of the ryanodine receptor protein. Bmax and KD of the chicken $[^3H]ryanodine$ binding sites were 12.52 pmol/mg protein and 14.53 nM, respectively. Specific $[^3H]ryanodine$ binding was almost maximal at $50{\sim}100$ ${\mu}M$ $Ca^{2+}$, but was not increased by 5 mM AMP and not inhibited by high $Ca^{2+}$. Binding was significantly inhibited by $20{\sim}100$ ${\mu}M$ ruthenium red and 1 mM tetracaine, but slightly inhibited by $Mg^{2+}$. From the above results, it is suggested that chicken SR vesicles have the ryanodine binding sites to which the binding of ryanodine is almost maximal at $50{\sim}10$ ${\mu}M$ $Ca^{2+}$, is significantly inhibited by ruthenium red and tetracaine, slightly inhibited by $Mg^{2+}$, but not affected by AMP and not inhibited by high $Ca^{2+}$.

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An Efficient Address Mapping Table Management Scheme for NAND Flash Memory File System Exploiting Page Address Cache (페이지 주소 캐시를 활용한 NAND 플래시 메모리 파일시스템에서의 효율적 주소 변환 테이블 관리 정책)

  • Kim, Cheong-Ghil
    • Journal of Digital Contents Society
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    • v.11 no.1
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    • pp.91-97
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    • 2010
  • Flash memory has been used by many digital devices for data storage, exploiting the advantages of non-volatility, low power, stability, and so on, with the help of high integrity, large capacity, and low price. As the fast growing popularity of flash memory, the density of it increases so significantly that its entire address mapping table becomes too big to be stored in SRAM. This paper proposes the associated page address cache with an efficient table management scheme for hybrid flash translation layer mapping. For this purpose, all tables are integrated into a map block containing entire physical page tables. Simulation results show that the proposed scheme can save the extra memory areas and decrease the searching time with less 2.5% of miss ratio on PC workload and can decrease the write overhead by performing write operation 33% out of total writes requested.

Extracellular synthesis of silver nanoparticle by Pseudomonas hibiscicola - Mechanistic approach

  • Punjabi, Kapil;Mehta, Shraddha;Yedurkar, Snehal;Jain, Rajesh;Mukherjee, Sandeepan;Kale, Avinash;Deshpande, Sunita
    • Advances in nano research
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    • v.6 no.1
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    • pp.81-92
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    • 2018
  • Biosynthesis of nanoparticles has acquired particular attention due to its economic feasibility, low toxicity and simplicity of the process. Extracellular synthesis of nanoparticles by bacteria and fungi has been stated to be brought about by enzymes and other reducing agents that may be secreted in the culture medium. The present study was carried out to determine the underlying mechanisms of extracellular silver nanoparticle synthesis by Pseudomonas hibiscicola isolated from the effluent of an electroplating industry in Mumbai. Synthesized nanoparticles were characterized by spectroscopy and electron microscopic techniques. Protein profiling studies were done using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (1D-SDS PAGE) and subjected to identification by Mass Spectrometry. Characterization studies revealed synthesis of 50 nm nanoparticles of well-defined morphology. Total protein content and SDS PAGE analysis revealed a reduction of total protein content in test (nanoparticles solution) samples when compared to controls (broth supernatant). 45.45% of the proteins involved in the process of nanoparticle synthesis were identified to be oxidoreductases and are thought to be involved in either reduction of metal ions or capping of synthesized nanoparticles.

The Development of Forest Fire Statistical Management System using Web GIS Technology

  • Jo, Myung-Hee;Kim, Joon-Bum;Kim, Hyun-Sik;Jo, Yun-Won
    • Proceedings of the KSRS Conference
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    • 2002.10a
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    • pp.183-190
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    • 2002
  • In this paper forest fire statistical information management system is constructed on web environment using web based GIS(Geographic Information System) technology. Though this system, general users can easily access forest fire statistical information and obtain them in visual method such as maps, graphs, and text if they have web browsers. Moreover, officials related to forest fire can easily control and manage all information in domestic by accessing input interface, retrieval interface, and out interface. In order to implement this system, IIS 5.0 of Microsoft is used as web server and Oracle 8i and ASP(Active Server Page) are used for database construction and dynamic web page operation, respectively. Also, Arc IMS of ESRI is used to serve map data using Java and HTML as system development language. Through this system, general users can obtain the whole information related to forest fire visually in real time also recognize forest fire prevention. In addition, Forest officials can manage the domestic forest resource and control forest fire dangerous area efficiently and scientifically by analyzing and retrieving huge forest data through this system. So, they can save their manpower, time and cost to collect and manage data.

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Studies on Viral Disease of masu salmon, Oncorhynchus masou-II Isolation of infectious hematopoietic necrosis virus form masu salmon fry (산천어의 바이러스성 질병에 관한 연구-II -산천어 치어에서 1HNV 분리-)

  • Sohn, Sang-Gyu;Park, Myoung-Ae;Park, Jeong-Woo
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.87-92
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    • 1993
  • In February of 1990, an epizootic disease to masu salmon. Onchorynchus masou cultured at the hatchery of trout in Samchuk. Kwangwondo have broken out and induced heavy mortality. An infectious hematopoietic necrosis virus(IHNV) was isolated from diseased masu salmon fry by the use of fish cell line, CHSE-214. This IHNV isolated from masu salmon was compared with USA isolates of IHNV, SRCV and RB-76 by analysis of virion proteins in sodium dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE) and neutralization tests with two monoclonal antibodies raised against SRCV(MAb SRCV/A4) and RB-76(MAb RB/B5). In the antigenicity and the size of structural proteins. this IHNV, SCS atrain was smilar to RB-76 belonged to the electropherotype I proposed by Hsu et al.(1986).

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Allergenic potential and enzymatic resistance of buckwheat

  • Lee, Sujin;Han, Youngshin;Do, Jeong-Ryong;Oh, Sangsuk
    • Nutrition Research and Practice
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    • v.7 no.1
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    • pp.3-8
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    • 2013
  • Buckwheat is known as a health food but is one of the major food allergens triggering potentially fatal anaphylaxis in Asia, especially in Japan and Korea. This study was conducted to investigate the characteristic of enzymatic resistance of buckwheat protein and allergenic potential. Enzymatic resistance of buckwheat protein was performed with in vitro digestibility test in simulated gastric fluid (SGF), pH 1.2, using pepsin and simulated intestinal fluid (SIF) using chymotrypsin. Reactivity of buckwheat proteins to human IgE was performed using six allergic patients sensitized to buckwheat. Buckwheat's IgE levels were measured using the Phadia UniCAP-system. Buckwheat protein, 16 kDa, still remained after 30 min treatment of pepsin on SDS-PAGE. Even though 16 kDa almost disappeared after 60 min treatment, two out of the six buckwheat patients' sera showed reactivity to hydrolysate after 60 min treatment, indicating that allergenicity still remained. In simulated intestinal fluid (SIF) using chymotrypsin, buckwheat protein, 24 kDa, showed resistance to hydrolysis with chymotrypsin on SDS-PAGE, and still had allergenicity based on the result of ELISA. Our results suggest that buckwheat proteins have strong resistance to enzyme degradation. This may be attributed in part to the allergenic potential of buckwheat. Further study should be continued regarding buckwheat allergy.