• Title/Summary/Keyword: G-BSA

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Partial characterization of a 29kDa cysteine protease purified from Taenia solium metacestodes

  • KIM Ji-Young;YANG Hyun-Jong;KIM Kwang-Sig;CHUNG Young-Bae
    • Parasites, Hosts and Diseases
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    • v.43 no.4 s.136
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    • pp.157-160
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    • 2005
  • A 29kDa cysteine protease of Taenia solium metacestodes was purified by Mono Q anion-exchanger and Superose 6 HR gel filtration chromatography. The enzyme was effectively inhibited by cysteine protease inhibitors, such as iodoacetic acid (IAA) and trans-epoxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) while inhibitors acting on serine- or metallo-proteases did not affect the enzyme activity. The purified enzyme degraded human immunoglobulin G (IgG), collagen and bovine serum albumin (BSA), but human IgG was more susceptible for proteolysis by the enzyme. To define the precise biological roles of the enzyme, more detailed biochemical and functional studies would be required.

Development of Radioimmunoassay(RIA) for Residue Analysis with Endosulfan in Water and Carp(Cyprinus carpio L.) (잉어중 Endosulfan의 잔류분석(殘留分析)을 위한 Radioimmunoassay(RIA)의 개발(開發))

  • Lee, Kang-Bong;Shim, Jae-Han;Suh, Yong-Tack
    • Korean Journal of Environmental Agriculture
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    • v.13 no.3
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    • pp.262-270
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    • 1994
  • The established methods in the residue analysis of endosulfan require an extensive sample clean-up prior to quantification by relatively complex equipment. A radioimmunoassay(RIA) provides a simple procedure with theoretically higher sensitivity and specificity necessitating only a minimum of sample clean-up. Endosulfan-specific antibodies were developed in rabbits by using a bovine serum albumin(BSA) conjugate wherein the alcohol form of endosulfan was multiply bound to the protein via succinylation. Produced antibodies showed the high titers to endosulfan-BSA(1 : 32,000). An RIA method was developed in water and carp by using $^{14}C-labeled$ endosulfan as a tracer. The lowest detection amount of endosulfan was 1 ng in the liver, kidneys, gut and water samples, and 3 ng in the whole body sample of carp without any clean-up, corresponding to 0.1 ppb of endosulfan.

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A Study for Effects of Magsungsinyeom-bang(Moxingshenyanfang) on Membranous Nephropathy induced by Cationic Bovine Serum Albuminin in Mouse Model(Effects of Magsungsinyeom-bang on Membranous Nephropathy) (막성신염방(膜性腎炎方)이 Cationized Bovine Serum Albumin투여로 유발된 Mouse의 Membranous Nephropathy에 미치는 영향)

  • Kim, Jae-Hyeong;Cho, Chung-Sik;Kim, Cheol-Jung
    • Korean Journal of Acupuncture
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    • v.24 no.4
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    • pp.163-180
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    • 2007
  • Objective : Oriental medicines have been applied to Membranous nephropathy(MN) for the purpose on increasing renal blood flow and modulating immune activity in according to Oriental medicine theories. Magsungsinyeom-bang (MSSYB) is one of the prescription which is known to show positive results in clinic with lack of laboratorial evidence. Thus, this study was aimed to evaluate the effects of MSSYB and partially investigate the mechanisms of it. Methods : The effect of MSSYB was evaluated by the morphology for the GBM thickening, protein excretion in urine and biochemical parameters in serum using cBSA-induced MN mice model. Mice were administered with MSSYB(250 or 500 mg/kg) or PBS for control group from experimental week 3 for 4 weeks. Results : 24 hrs proteinuria and the concentrations BUN was significantly decreased in the MS groups compared to the control group while the concentrations of serum albumin was higher in the MS groups than control group. MSSYB didn't affect the ratio of CD3e+/CD or 19CD4+/CD8 in the spleen and kidney, but inhibit the expression of IL-1${\beta}$, TNF-${\alpha}$, IL-6, and production of IgG and IgM. In histological analysis of kidney tissue, thickening of GBM was significantly decreased in the MS group compared to control group. Conclusions : MSSYB showed the positive results on the cBSA-induced membranous nephropathy in mice, thus, it could be a useful candidate for oriental drug for treating the membranous nephropathy in clinic.

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Effect of Defined KSOM Medium on the Development of 1-antitrypsin Transgenic Nuclear Transfer Bovine Embryos

  • M.M.U. Bhuiyan;J.K. Cho;G. Jang;Park, E.S.;S.K. Kang;Lee, B.C.;W.S. Hwang
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.74-74
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    • 2002
  • Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

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Survival and Development of Porcine Embryos Produced in vitro Using Open Pulled Straw Methods (돼지에서 Open Pulled Straw(OPS) 방법에 의해 동결-융해한 수정란의 생존능력)

  • Lee, S.Y.;Yu, J.S.;Sa, S.J.;Park, C.K.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.255-262
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    • 2006
  • The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

Syntheses of 3-Pyrimidyl- and 3-Pyranyl-5,6-benzocoumarin Derivatives

  • El-Deen, Ibrahim M.;Al-Wakeel, El-Sayed I.;El-Mawla, Ahmed G.
    • Bulletin of the Korean Chemical Society
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    • v.23 no.4
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    • pp.610-612
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    • 2002
  • A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of organophosphorus insecticide cyanophos. An analogue (hapten) of cyanophos was synthesized and was coupled to BSA to produce polyclonal antibodi es from rabbits. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 310 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivities with other organophosphorus pesticides except for parathion-methyl, which makes the assay suitable for the selective detection of cyanophos.

FET type Urea Sensor (FET형 요소 감지 소자)

  • Moon, Byung-Joon;Lee, Jong-Hyun;Sohn, Byung-Ki
    • Proceedings of the KIEE Conference
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    • 1987.07a
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    • pp.490-492
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    • 1987
  • Urea-Sensor was fabricated by immobilizing urease on ISFET's gate using BSA(bovine serum albumin) and glutaraldehyde, and its characteristics were examined. This sensor showed approximately linear characteristic in the urea concentration range of $3{\times}10^{-5}-10^{-9}$ (g/ml). Fast response time was obtained and minute amounts of expensive enzyme were used in comparison to general electrode type biosensors.

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Patterning Biological Molecules onto Poly(amidoamine) Dendrimer on Gold and Glass

  • Hong, Mi-Young;Lee, Do-Hoon;Yoon, Hyun C.;Kim, Hak-Sung
    • Bulletin of the Korean Chemical Society
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    • v.24 no.8
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    • pp.1197-1202
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    • 2003
  • Patterning of biological molecules was attempted on both gold and glass using fourth generation (G4) poly(amidoamine) (PAMAM) dendrimer as an interfacing layer between solid surfaces and biomolecules. As for the patterning of avidin and anti-biotin antibody on gold, PAMAM dendrimers representing amine functionalities were firstly printed onto the 11-mercaptoundecanoic acid SAM by microcontact printing, followed by biotinylation, and reacted with fluorescence-labeled avidin or anti-biotin antibody. Fluorescence microscopic analysis revealed that the patterns of avidin and anti-biotin antibody were well constructed with the resolution of < 2 ㎛. The PAMAM dendrimers were also printed onto aldehyde-activated slide glass and reacted directly with anti-BSA antibodies, which had been oxidized with sodium periodate. As a result, distinct patterns of the anti-BSA antibodies were also obtained with a comparable edge resolution to that of avidin patterns on gold. These results clearly show that PAMAM dendrimers can be adopted as an interfacing layer for the patterning of biological molecules on solid surfaces with micrometer resolution.

Protein Adsorption and Hydrodynamic Stability of a Dense, Pellicular Adsorbent in High-Biomass Expanded Bed Chromatography

  • Chow, Yen Mei;Tey, Beng Ti;Ibrahim, Mohd Nordin;Ariff, Arbakariya;Ling, Tae Chuan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.268-272
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    • 2006
  • A dense, pellicular UpFront adsorbent ($p=1.5 g/cm^3$, UpFront Chromatography, Cophenhagen, Denmark) was characterized in terms of hydrodynamic properties and protein adsorption performance in expanded bed chromatography. Cibacron Blue 3GA was immobilised into the adsorbent and protein adsorption of bovine serum albumin (BSA) was selected to test the setup. The Bodenstein number and axial dispersion coefficient estimated for this dense pellicular adsorbent was 54 and $1.63{\times}10^{-5}m^2/s$, respectively, indicating a stable expanded bed. It could be shown that the BSA protein was captured by the adsorbent in the presence of 30% (w/v) of whole-yeast cells with an estimated dynamic binding capacity $(C/C_o = 0.01)$ of approximately 6.5 mg/mL adsorbent.

A Possible Merge of FRET and SPR Sensing System for Highly Accurate and Selective Immunosensing

  • Lee, Jae-Beom;Chen, Hongxia;Lee, Jae-Wook;Sun, Fangfang;Kim, Cheol-Min;Chang, Chul-Hun L.;Koh, Kwang-Nak
    • Bulletin of the Korean Chemical Society
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    • v.30 no.12
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    • pp.2905-2908
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    • 2009
  • Immuno-sensing for high accurate and selective sensing was performed by fluorescence spectroscopy and surface plasmon resonance (SPR), respectively. Engineered assembly of two fluorescent quantum dots (QDs) with bovine serum albumin (BSA) and anti-BSA was fabricated in PBS buffer for fluorescence analysis of fluorescence resonance energy transfer (FRET). Furthermore, the same bio-moieties were immobilized on Au plates for SPR analysis. Naturally-driven binding affinity of immuno-moieties induced FRET and plasmon resonance angle shift in the nanoscale sensing system. Interestingly, the sensing ranges were uniquely different in two systems: e.g., SPR spectroscopy was suitable for highly accurate analysis to measure in the range of 10$^{-15{\sim}-10$ng/mL while the QD fluorescent sensing system was relatively lower sensing ranges in 10$^{-10{\sim}-6$ng/mL. However, the QD sensing system was larger than the SPR sensing system in terms of sensing capacity per one specimen. It is, therefore, suggested that a mutual assistance of FRET and SPR combined sensing system would be a potentially promising candidate for high accuracy and reliable in situ sensing system of immune-related diseases.