• Title/Summary/Keyword: N2a cells

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Antigen analysis of Toxoplasma gondii Iysate and excretory-secretory materials by enzyme-linked immunoelectrotransfer blot (EITB) (효소면역 전기영동이적법에 의한 톡소포자충 용해물 및 분비 항원의 분석)

  • 안명희;손혁진
    • Parasites, Hosts and Diseases
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    • v.32 no.4
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    • pp.249-258
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    • 1994
  • Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

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Resolving the 'Gray sheep' Problem Using Social Network Analysis (SNA) in Collaborative Filtering (CF) Recommender Systems (소셜 네트워크 분석 기법을 활용한 협업필터링의 특이취향 사용자(Gray Sheep) 문제 해결)

  • Kim, Minsung;Im, Il
    • Journal of Intelligence and Information Systems
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    • v.20 no.2
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    • pp.137-148
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    • 2014
  • Recommender system has become one of the most important technologies in e-commerce in these days. The ultimate reason to shop online, for many consumers, is to reduce the efforts for information search and purchase. Recommender system is a key technology to serve these needs. Many of the past studies about recommender systems have been devoted to developing and improving recommendation algorithms and collaborative filtering (CF) is known to be the most successful one. Despite its success, however, CF has several shortcomings such as cold-start, sparsity, gray sheep problems. In order to be able to generate recommendations, ordinary CF algorithms require evaluations or preference information directly from users. For new users who do not have any evaluations or preference information, therefore, CF cannot come up with recommendations (Cold-star problem). As the numbers of products and customers increase, the scale of the data increases exponentially and most of the data cells are empty. This sparse dataset makes computation for recommendation extremely hard (Sparsity problem). Since CF is based on the assumption that there are groups of users sharing common preferences or tastes, CF becomes inaccurate if there are many users with rare and unique tastes (Gray sheep problem). This study proposes a new algorithm that utilizes Social Network Analysis (SNA) techniques to resolve the gray sheep problem. We utilize 'degree centrality' in SNA to identify users with unique preferences (gray sheep). Degree centrality in SNA refers to the number of direct links to and from a node. In a network of users who are connected through common preferences or tastes, those with unique tastes have fewer links to other users (nodes) and they are isolated from other users. Therefore, gray sheep can be identified by calculating degree centrality of each node. We divide the dataset into two, gray sheep and others, based on the degree centrality of the users. Then, different similarity measures and recommendation methods are applied to these two datasets. More detail algorithm is as follows: Step 1: Convert the initial data which is a two-mode network (user to item) into an one-mode network (user to user). Step 2: Calculate degree centrality of each node and separate those nodes having degree centrality values lower than the pre-set threshold. The threshold value is determined by simulations such that the accuracy of CF for the remaining dataset is maximized. Step 3: Ordinary CF algorithm is applied to the remaining dataset. Step 4: Since the separated dataset consist of users with unique tastes, an ordinary CF algorithm cannot generate recommendations for them. A 'popular item' method is used to generate recommendations for these users. The F measures of the two datasets are weighted by the numbers of nodes and summed to be used as the final performance metric. In order to test performance improvement by this new algorithm, an empirical study was conducted using a publically available dataset - the MovieLens data by GroupLens research team. We used 100,000 evaluations by 943 users on 1,682 movies. The proposed algorithm was compared with an ordinary CF algorithm utilizing 'Best-N-neighbors' and 'Cosine' similarity method. The empirical results show that F measure was improved about 11% on average when the proposed algorithm was used

    . Past studies to improve CF performance typically used additional information other than users' evaluations such as demographic data. Some studies applied SNA techniques as a new similarity metric. This study is novel in that it used SNA to separate dataset. This study shows that performance of CF can be improved, without any additional information, when SNA techniques are used as proposed. This study has several theoretical and practical implications. This study empirically shows that the characteristics of dataset can affect the performance of CF recommender systems. This helps researchers understand factors affecting performance of CF. This study also opens a door for future studies in the area of applying SNA to CF to analyze characteristics of dataset. In practice, this study provides guidelines to improve performance of CF recommender systems with a simple modification.

  • Quantitative Measurement of Membrane $Na^+-K^+$ ATPase Activity using Thallium-201 : Comparison with Rubidium-86 (Thallium-201을 이용한 세포막 $Na^+-K^+$ ATPase 활성도 측정: Rubidium-86 측정법과의 비교)

    • Lee, Jae-Tae;Lee, In-Kyu;Sohn, Sang-Kyun;Lee, Kyu-Bo
      • The Korean Journal of Nuclear Medicine
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      • v.32 no.2
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      • pp.121-128
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      • 1998
    • Purpose: $Na^+-K^+$ ATPase activity has been estimated by the degree of inhibition of cation transport by cardiac glycosides (ouabain) using Rb-86 as a substrate. The biological characteristisc of T1-201 is known to be similar to those of potassium as a transport substrate in the presence of glucose, insulin or phobol myristate acetate (PMA). The purpose of this study was to measure ouabain sensitive $Na^+-K^+$ ATPase activity using T1-201 and compare with that using Rb-86. Materials and Methods: Smooth muscle cells isolated from rat aorta or human placental umbilical artery were cultured, and used to measure cellular $Na^+-K^+$ ATPase activity. $Na^+-K^+$ ATPase activity was measured as a percentage decrease in cellular uptake of T1-201 or Rb-86 by ouabain under the presence of glucose, insulin or PMA in media. Results: $Na^+-K^+$ ATPase activity measured with T1-201, as a transport substrate, was not different from those measured with Rb-86 in rat or human smooth muscle cell preparation. Incubation with high concentration glucose resulted in about 30% decrease in enzyme activity. In contrast, insulin or PMA resulted in 10-70% or 28% increases from baseline activity, respectively. Conclusion: These results suggests that 71-201 could replace Rb-86 in measurement of ouabain sensitive $Na^+-K^+$ ATPase activity in vitro. High level of glucose concentration decreased cellular $Na^+-K^+$ ATPase activity, but insulin or PMA increased it.

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    Growth enhancement and cytotoxicity of Korean mistletoe fractions on human cell lines (한국산 겨우살이 분획물의 면역세포의 생육증진 및 세포독성)

    • Lee, So-Jin;Lee, Mi-Kyoung;Choi, Geun-Pyo;Yu, Chang-Yeon;Roh, Seong-Kyu;Kim, Jong-Dai;Lee, Hyeon-Yong;Lee, Jin-Ha
      • Korean Journal of Medicinal Crop Science
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      • v.11 no.1
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      • pp.62-70
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      • 2003
    • The biological activities on human immune and cancer cell lines of the four kinds of Korean mistletoes (Korean Viscum album, var. coloratum, : Korean Viscum sp. in Quercus acutissima Carr., Korean Viscum sp. in Castanea crenata, Korean Viscum sp. in Betula platyphylla, and Korean Viscum sp. in Salix koreensis) extracts were investigated. The extracts were preparated with ethanol, and fractionated with n-butanol, ethyl acetate, chloroform, hexane, and second distilled water. Cytotoxic potencies of the fractions on human normal lung cell line (HEL 299) showed under 28% in the concentration of 0.5 mg/ml. Growth inhibition effect of the Korean mistletoe extracts on the several human cancer cell lines depends on the concentration of the extracts, and extracting solvent. The hexane, chloroform, and ethyl acetate fractions indicated a strong anticancer activity, but not in aqueous and butanol fractions. Some mistletoe fractions have a different characteristic on the cancer cell lines. Stimulation on the growth of human immuno cell lines(B cell : Raji, T cell: Jurkat) of the extracts were confirmed in the ethyl acetate, chloroform, hexane fractions, but not in aqueous system.

    Multiplication of Infectious Flacherie and Densonucleosis Viruses in the Silkworm, Bombyx mori (가잠의 전염성 연화병 및 농핵병 바이러스 증식에 관한 연구)

    • 김근영;강석권
      • Journal of Sericultural and Entomological Science
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      • v.25 no.2
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      • pp.1-31
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      • 1984
    • Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

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