• Title/Summary/Keyword: G-BSA

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THE EFFECT OF SUPEROXIDE DISMUTASE ON EXPERIMENTAL GINGIVITIS AND ACTIVITY OF 3T3 FIBROBLAST (Superoxide Dismutase가 백서의 실험적 치은염과 3T3 섬유모 세포의 활성에 미치는 영향)

  • Kim, Yoon-Seong;Yoo, Hyung-Keun;Kang, Hyun-Ku;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.222-238
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    • 1995
  • Inflammatory cells may produce active species of oxygen in antimicrobial defense. While such species can directly damage surrounding tissue, their major secondary role may be to mediate important components of the inflammatory response. Superoxide dismutase, antioxidant, have significant anti-inflammatory properties in rheumatoid arthritis, ischemic tissue injury and gastrointestinal disease. Increased oxidative product formation diseases. And superoxide dismutase produced by Porphyromonas Gingivalis is resistant to killing by polymorphonuclear leukocyte. The purpose of this study was to investigate on the effects of superoxide dismutase in 3T3 fibroblast and in experimental gingivitis in the rats. The effect of superoxide dismutase(SOD) to cell morphology and cell activity was measured in cultured mouse 3T3 fibroblast. After experimental gingivitis were induced by lipopolysaccharide(LPb) and bovine serum albumin(BSA), injection of SOD were done. WBC count and histologic findings were observed at 1, 2, 3, and 7 days. The results were as follows; 1. There was a little difference between LPS treated groups and SOD treated groups in 3T3 fibroblast morpholoy. 2. There was no difference between only SOD treated groups (except SOD 150U at 3days) and control in 3T3 fibroblast activity. 3. LPS $0.5{\mu}g/ml$ and SOD treated groups (except 150U) had decreased 3T3 fibroblast activity and no significant difference at 3 days. 4. LPS $5.0{\mu}g/ml$ and SOD treated groups were significantly increased cell activity of 3T3 fibroblast than control group at 1 day(P<0.05). 5. In LPS induced gingivitis, the number of leukocytes in SOD treated was significantly decreased than in saline treated at 1 day(P<0.05). 6. In histopathologic findings of LPS or BSA induced gingivitis, inflammatorycell infiltration in SOD treated groups were less than in saline treated group at 1, 2 and 3 days.

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Enzyme-Linked Immunosorbent Assay for Detection of Nivalenol (Nivaleno의 검출을 위한 효소 면역 측정법)

  • 손동화;이향범;곽보연;김수호;권창희
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.129-134
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    • 1998
  • To develop an enzyme-linked immunosorbent assay (ELISA) for nivalenol (NIV), we produced polyclonal antibodies against tetraacetyl nivalenol (Ac4-NIV) and established ELISA conditions. Ac4-NIV-hemisuccinate conjugated to bovine serum albumin (Ac4-NIV-HS-BSA) was immunized with Freund's adjuvants into rabbits subcutaneously several times. By use of the antiserum showing the highest titer and Ac4-NIV-HS-HRP conjugate, we established competitive direct ELISA (cdELISA). Standard curve of cdELISA showed that the detection range of Ac4-NIV was about 10~5,000 ng/ml (ppb). The cross-reactivities of the polyclonal antibody towards Ac4-NIV and acetyl T-2 were 100 and 70% respectively, and those towards NIV, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, triacetyl deoxynivalenol, fusarenonX, and T-2 were less than 0.1%. When cdELISA was applied to NIV-spiked corns followed by extraction with 70% acetonitrile and acetylation with acetic anhydride in pyridine, the recovery rates of the Ac4-NIV were 108, 143, and 70% (average, 107%) in the levels of 100, 300, and 1,000 ng/g (ppb), respectively.

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Development of Miniaturized Automatic Chromatography System for validation Study of Chromatographic Resin lifetime (크로마토그래피 담체의 수멍을 검증하기 위한 자동화 미니 크로마토그래피 시스템 개발)

  • 박재하;서창우
    • KSBB Journal
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    • v.17 no.4
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    • pp.326-332
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    • 2002
  • The quality of biopharmaceutical proteins is strongly affected by a manufacturing process employed to produce Et, and thus validation of the manufacturing bioprocess is a very important issue. Chromatography is probably the most widely used bioprocess unit operation for protein purification. In this study, a miniaturized automatic chromatography system was designed and constructed for scale-down studies for process chromatography validation. This system, named MiniValChrom, has the following features: automatic and repeated operation, flexible sequences and intervals among the steps, on-line and real-time monitoring and control, method files savings, etc. Using the MiniValChrom, we peformed a case study of an abbreviated experiment to estimate chromatographic resin lifetime. BSA (bovine serum albumin) and Cibacron Blue 3G-A were used as the model protein and the resin, respectively. The resin deterioration was evaluated by determining and monitoring the HETP and NTP values from the chromatograms every 5 cycles. It was observed that the HETP and the NTP values were changed by 9% after 15 cycles. The resin lifetime validation could be completed by repeating this experiment until the HETP value reached a predetermined value. The MiniValChrom's concept and the protocol suggested in this study can serve as a rapid and economical tool for the validation studies of bioprocess chromatography system.

A fiber optic surface plasmon resonance (SPR) sensorusing cyclic olefin copolymer (COC) polymer prism (Cyclic olefin copolymer (COC) 폴리머 프리즘을 사용한 광섬유 기반 표면 플라즈몬 공명 (SPR) 바이오 센서)

  • Yun, Sung-Sik;Lee, Soo-Hyun;Ahn, Chong-H.;Lee, Jong-Hyun
    • Journal of Sensor Science and Technology
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    • v.17 no.5
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    • pp.369-374
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    • 2008
  • A novel fiber optic surface plasmon resonance (SPR) sensor using cyclic olefin copolymer (COC) prism with the spectral modulation is presented. The SPR sensor chip is fabricated using the SU-8 photolithography, Ni-electroplating and COC injection molding process. The sidewall of the COC prism is partially deposited with Au/Cr (45/2.nm thickness) by e-beam evaporator, and the thermal bonding process is conducted for micro fluidic channels and optical fibers alignment. The SPR spectrum for a phosphate buffered saline (0.1.M PBS, pH.7.2) solution shows a distinctive dip at 1300.nm wavelength, which shifts toward longer wavelength with respect to the bovine serum albumin (BSA)concentrations. The sensitivity of the wavelength shift is $1.16\;nm{\cdot}{\mu}g^{-1}{\cdot}{\mu}l^{-1}$. From the wavelength of SPR dips, the refractive indices (RI) of the BSA solutions can be theoretically calculated using Kretchmann configuration, and the change rate of the RI was found to be $2.3{\times}10^{-5}RI{\cdot}{\mu}g^{-1}{\cdot}l^{-1}$. The realized fiber optic SPR sensor with a COC prism has clearly shown the feasibility of a new disposable, low cost and miniaturized SPR biosensor for biochemical molecular analyses.

Application of Silver Ion for Clinical Mastitis in Holstein Cows (임상형 유방염을 가진 홀스테인(Holstein) 소에 대한 은 이온의 적용 연구)

  • Seol, Jae-Won;Hur, Tai-Young;Jung, Young-Hun;Park, Sang-Youel;Kang, Seog-Jin
    • Journal of Veterinary Clinics
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    • v.27 no.3
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    • pp.246-251
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    • 2010
  • Mastitis is an inflammation of the mammary gland that develops as a response to pathogenic microorganisms. Somatic cell count (SCC) and bacteriological analysis are the most accepted tools to determine udder health. This study evaluated the effects of intra-mammary gland infusion of silver ion against clinical mastitis in Holstein cows. Silver ion (20 ${\mu}g$/ml) was infused through the intra-mammary gland in quarters having clinical mastitis and milk was collected to determine SCC, and levels of lactoferrin and bovine serum albumin (BSA). Silver ion infusion decreased udder inflammation, firmness and swelling, and reduced clots, BSA, lactoferrin and SCC in milk. However, milk yield and circulating blood cells remained unchanged. The silver ion-mediated reductions of BSA and SCC indicate reduced inflammation and bacterial activity in silver ion-treated mammary glands in Holsteins with mastitis, which may be exploited in a curative strategy.

Development of Serum-free Media for the Culture of Mouse Hybridoma (I) ; Determination of Optimal Media Composition (쥐 하이브리도마 세포배양을 위한 무혈청 배지개발( I ) -최적 배지성분의 결정-)

  • 조보연;최태부
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.481-488
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    • 1989
  • A serum-free medium that could be used for the large-scale culture of mouse hybridoma to produce monoclonal antibodies was developed. The medium was based on a 1:1 mixture of Iscove's Modified Dulbecco's Medium and Ham's F-12, supplemented with insulin 10$\mu\textrm{g}$/$m\ell$, transferrin 10$\mu\textrm{g}$/$m\ell$, ethanolamine 10$\mu$M and selenium 30nM (designated EBM (enriched basal medium) with the supplements). The effect of various supplements of steroid hormones, vitamins, lipid and mineral salts was investigated and their optimal concentration was determined to replace fetal calf serum (PCS). These components were added respectively and then added by way of two or three combination to discern of which component combination was effective to the culture of hybridoma. As a result, serum-free medium KM3 (EBM with BSA 100$\mu\textrm{g}$/$m\ell$, mineral cocktail and 0.05% PEG) was deter-mined. The hybridoma Alps 25-3 cultured in this medium showed almost the same growth rate as in medium added with 2% fetal bovine serum. However, the antibody concentration from KM3 cultures was 80% of that obtained from culture with FCS. KM3 was also examined for the culture of other mouse hybridomas, KW, A4W & HCGK, and it was confirmed that it could support the growth of these hybridomas and the production of monoclonal antibodies.

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Electrochemical Immunosensor Using a Gas Diffusion Layer as an Immobilization Matrix

  • Kim, Yong-Tae;Oh, Kyu-Ha;Kim, Joo-Ho;Kang, Hee-Gyoo;Choi, Jin-Sub
    • Bulletin of the Korean Chemical Society
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    • v.32 no.6
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    • pp.1975-1979
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    • 2011
  • The modification of a gas diffusion layer (GDL), a vital component in polymer electrolyte fuel cells, is described here for use in the electrochemical detection of antibody-antigen biosensors. Compared to other substrates (gold foil and graphite), mouse anti-rHBsAg monoclonal antibody immobilized on gold-coated GDL (G-GDL) detected analytes of goat anti-mouse IgG antibody-ALP using a relatively low potential (-0.0021 V vs. Ag/AgCl 3 M NaCl), indicating that undesired by-reactions during electrochemical sensing should be avoided with G-GDL. The dependency of the signal against the concentration of analytes was observed, demonstrating the possibility of quantitative electrochemical biosensors based on G-GDL substrates. When a sandwich method was employed, target antigens of rHBsAg with a concentration as low as 500 ng/mL were clearly measured. The detection limit of rHBsAg was significantly improved to 10 ng/mL when higher concentrations of the 4-aminophenylphosphate monosodium salt (APP) acting on substrates were used for generating a redox-active product. Additionally, it was shown that a BSA blocking layer was essential in improving the detection limit in the G-GDL biosensor.

Immunoelectron Microscopic Study on the Paneth Cell of Rabbit after the Common Bile Duct Ligation (총담관결찰후 집토끼 Paneth세포의 변화에 대한 면역전자현미경적 연구)

  • Park, Kyung-Ho;Cho, Hwee-Dong;Yang, Nam-Gil;Ahn, E-Tay;Ko, Jeong-Sik;Kim, Jin-Gook
    • Applied Microscopy
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    • v.24 no.2
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    • pp.78-92
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    • 1994
  • Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

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Anti-allergic Activity of Oriental Medicinal Herbs

  • Kim, Young-Mi
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.5
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    • pp.282-288
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    • 2006
  • The effects of extracts from various Oriental medicinal herbs on mast-cell-mediated allergic reactions were investigated in this study. Out of these extracts, the medicinal herb Atractylodis rhizoma alba exhibited the most potent activity in the cells, with $IC_50$ values of $20.5\;{\mu}g/ml$, by DNP-BSA and $33.04\;{\mu}g/ml$, by compound 48/80. Similar to the in vitro activity, Atractylodis rhizoma alba inhibited compound-48/80-induced systemic anaphylaxis by 31.3% at 300 mg/kg in mice. It also suppressed the secretion of $TNF-{\alpha}$, a major pro-inflammatory cytokine in the cells. These results may suggest that Atractylodis rhizoma alba shows anti-allergic activity in compound 48/80-induced anaphylactic mice through the inhibition of mast cells.

Effect of Maternal Food Intake on the Specific Antibody Level to Dietary Antigens in Human Breast Milk (임산부의 식이섭취가 모유 중에 함유된 식이 단백질에 특이적인 항체 수준에 미치는 영향)

  • Jang, Yeong-Ae;Kim, Yeong-Na;Kim, Sun-Mi
    • The Korean Journal of Food And Nutrition
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    • v.8 no.2
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    • pp.70-78
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    • 1995
  • We had examined the levels of specific IgG and IgA to dietary antigens in human breast milk and the relationships between the maternal food intake and the specific antibody level. The highest antibody titers were found in colostrum and decreased as lactation progressed. The specific antibody level was not affected by maternal calorie or protein intake, but affected by the intake frequency of a kind of food. Egg and meat intake significantly related to anti-OVA IgG and anti-BSA IgA antibodies, respectively. Meat intake frequency was generally affected by the other specific antibody levels.

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