• Title/Summary/Keyword: G-BSA

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In Vitro Production of Bovine Embryos by Modification of Simple Defined Culture Medium (단순한정배양액의 성분조정에 의한 소 수정란의 체외생산)

  • 노상호;윤종택;한기영;이병천;황우석
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.235-243
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    • 1998
  • In this study, we investigated the effects of three kinds of culture medium (Charles and Rosenkrans; CRlaa, Tyrode's; TALP, synthetic oviduct fluid: SOF), insulin transferrin + selenium complex (ITS), macromolecules(polyvinyl alcohol: PVA, fetalb-ovine serum: FBS) and NaCl on the development of early bovine embryos. In experiment 1, there were no differences in embryo development among three kinds of embryo culture medium (CR $l_{aa}$ , TALP, SOF). In experiment 2, BSA, FBS and PVA were added each in TALP as macromolecule sources. The developmental rates of embryos in BSA or FBS added TALP were significantly higher than in PVA added one (p〈0.01), but there was no difference between BSA and FBS added groups. In experiment 3, bovine embryos were cultured in TALP with the following supplements: BSA alone(1, 3 or 8 mg/ml, each) or BSA(1, 3 or 8 mg/ml, each)+ITS (10$\mu\textrm{g}$/m1 insulin, 5 $\mu\textrm{g}$/ml transferrin, 5 ng/ml selenium). In higher concentration of BSA and ITS supplemented groups, the developmental rates over compacted morula were higher than others, but there was a significant effect of ITS only in 1 mg/ml of BSA added group (p〈0.05). In experiment 4, the effect of reduced concentration of NaCl was evaluated. The developmental rate over compacted morula in the medium containing 90 mM of NaCl was higher than in 114 mM group (p〈0.05). In conclusion, BSA could be used as a macromolecule source in bovine embryo culture, and ITS, as a serum substitute, could be used for improving of embryonic development. Also, reduction of NaCl concentration from 114 mM to 90 mM may improve the development of bovine embryos.bryos.

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Developmental Potential of Bovine Nuclear Transfer Embryos Cultured in Serum-free Medium

  • Lee, S. K;Kim, D. H.;G. S. Im;B. C. Yang;Park, H. S.;W. K. Chang;Lee, H. T.
    • Korean Journal of Animal Reproduction
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    • v.27 no.4
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    • pp.339-347
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    • 2003
  • The purpose of this study was to investigate the development of bovine nuclear transfer (NT) embryos cultured in serum-free conditions. Bovine NT embryos cultured in various culture conditions were compared blastocyst development, total cell number and apoptosis using TUNEL assay. In experiment 1, blastocyst rates of NT embryos were significantly higher (P<0.01) in FBS (22.0%) and BSA (26.6%) groups than in PVA (6.3%) group. Total cell number was significantly higher in FBS (78.4$\pm$19.4) and BSA (90.9$\pm$29.1) groups than in PVA group (46.0$\pm$0.0). Apoptotic cell number was significantly fewer in FBS (3.1$\pm$1.4) and BSA (1.7$\pm$1.4) groups than in PVA group (7.0$\pm$20.0) However, all of results were not different between the FBS and BSA group. In experiment 2, blastocyst rates of NT embryos were significantly higher (P<0.05) in fatty acid free-BSA (FAF-BSA) group (26.8%) than in fraction V-BSA group (11.2%). Total cell number were somewhat higher in FAF-BSA group (89.8$\pm$30.7) than in fraction V-BSA group (88.1$\pm$19.3). Apoptotic cell number were somewhat fewer in FAF-BSA (1.7$\pm$1.5) group than in fraction V-BSA group (4.2$\pm$2.9). These findings suggest that serum free condition were effective for the in vitro development of bovine NT embryos. Therefore, we concluded that fatty acid free-BSA has beneficial effect in development bovine NT embryos and can be use as a serum substitute.

Preparation for Protein Separation of an Ion-Exchange Polymeric Stationary Phase Presenting Amino Acid and Amine Units Through Surface Graft Polymerization

  • Choi Seong-Ho;Lee Kwang-Pill;Shin Chang-Ho
    • Macromolecular Research
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    • v.13 no.1
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    • pp.39-44
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    • 2005
  • Ion-exchange polymeric stationary phases presenting amino acid and amino groups were prepared by the surface grafting of glycidyl methacrylate onto a silica gel surface and subsequent amination. Three kinds of amino acids-L-arginine (Arg), D-lysine (Lys), and D-histine (His)-were used in this study. An ion-exchange polymeric stationary phase presenting ethylene diamine (EDA) was also prepared by surface graft polymerization. Separation of the model proteins bovine serum albumin (BSA), chick egg albumin (CEA), and hemoglobin (Hb) was performed using the amino acid- and amine-derived columns. In separating the CEA/BSA mixture, the resolution time of BSA was longer than that of CEA when using the EDA column, whereas the resolution time of BSA was shorter than that of CEA when using the Arg, Lys, and His columns. In the separation of the Hb/BSA mixture, the resolution time of BSA was longer than that of Hb in the EDA column, whereas the resolution time of BSA was shorter than that of Hb in the amino acid columns (D-Lys, L-Arg, and D-His).

A Study on the Effects of Chungyeolmaksungbang on Mouse Model of Membranous Nephropathy Induced by Cationic Bovine Serum Albumin (청열막성방(淸熱膜性方)이 Cationized Bovine Serum Albumin투여로 유발된 Mouse의 Membranous Nephropathy에 미치는 영향)

  • Choi, In-Gu;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.104-116
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    • 2008
  • Objective : Membranous nephropathy (MN) is the most common cause of adult nephrotic syndrome worldwide. MN has been defined as granular subepithelial deposition of IgG immune complexes along the glomerular basement membrane (GBM). We aimed to identify the effects of Chungyeolmaksungbang (CYMSB) treatment on cBSA-induced in MN mouse model. Methods : The effect of Chungyeolmaksungbang treatment was studied on the morphology and protein excretion in the cationized bovine serum albumin (cBSA)induced mouse chronic serum sickness nephritis model. One group of mice was given intra-peritoneal (i.p.) immunizing doses of cBSA and complete Freund's adjuvant. One week later, these animals began a single i.p. injection of cBSA for 4 weeks. A second group followed the same injection protocol, but was given CYMSB p.o. Results : Proteinuria significantly was decreased and serum albumin was increased in the group treated with cBSA and CYMSB extract compared with the control. Serum BUN was significantly decreased on CYMSB compared with control. CD3e+/CD19 cells ratio of peripheral blood was decreased and CD4+/CD8 cells was increased. Level of $IL-1{\beta}$ was significantly decreased, and $IFN-{\gamma}$ was significantly increased. Concentration of IgG and IgM was significantly decreased compared with control. Thickness of GBM was decreased on histological analysis of kidney. Deposition of CD4 and CD8 was decreased on immunohistochemical staining of kidney. Conclusions : We conclude that CYMSB treatment may could be a useful remedy agents for treating the MN with cBSA.

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Effect of the Crude Polysaccharides Fraction from Eleutherococcus senticosus as a Immunoadjuvant to Soluble Antigens (BSA and OVA) (오가피 조다당의 단백질 항원 (BSA and OVA)에 대한 면역증강효과)

  • 황수현;하은숙;유광원;신광순;이상훈;이재권;이경호;윤택준;박우문
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.167-175
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    • 2003
  • Eleutherococcus senticosus is a typical oriental folk medicinal herb. It has been used clinically as a anti-rheumatic disease, anti-stress, ischemic heart disease and gastric ulcer. In the present study, we examined the adjuvant activity of the crude polysaccharides fraction from Eleutherococcus senticosus, EN-3, on the induction of humoral and cellular immune responses against bovine serum albumin (BSA) or ovalbumin (OVA). The thioglycollate-induced macrophages and silica-induced dendritic-like cells cultured with BSA and EN-3 synergistically increased the production of TNF-$\alpha$ and IL-12. When mice were subcutaneously immunized with BSA + EN-3, the antibody titer against BSA was showed significantly higher than those immunized with BSA alone. In addition, when mice were immunized with OVA + FIA + EN-3, the antibody titer was showed similar patterns with the FCA. The assay for determining subisotype of antibody revealed that EN-3 augmented OVA-specific antibody titer of IgG1 and IgG2b. The culture supernatant obtained from splenocytes of mice treated with OVA + FIA + EN-3 also showed a higher level of both OVA-specific Th1-type (IL-2, IFN-${\gamma}$ and GM-CSF) and Th2-type cytokine (IL-4, IL-6 and IL-10). In vitro analysis of T cell proliferation to OVA on 8 weeks, the splenocytes of mice treated with OVA + EN-3 showed a significantly higher proliferating activity than those treated with OVA alone. These results suggest that EN-3 may possess adjuvant activities to potentially to enhance humoral as well as cellular immune response.

Experimental investigation of organic fouling mitigation in membrane filtration and removal by magnetic iron oxide particles

  • Jung, Jaehyun;Sibag, Mark;Shind, Bora;Cho, Jinwoo
    • Membrane and Water Treatment
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    • v.11 no.3
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    • pp.223-229
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    • 2020
  • Here magnetic iron oxide particles (MIOPs) were synthesized under atmospheric air and which size was controlled by regulating the flow rate of alkali addition and used for efficient removal of bovine serum albumin (BSA) from water. The MIOPs were characterized using field-emission scanning electron microscopy (FE-SEM), Fourier transformation-Infrared spectroscopy (FT-IR) and vibrating sample magnetometer (VSM). The results revealed a successful preparation of the MIOPs. The removal efficiency for BSA using MIOPs was found to be about 100% at lower concentrations (≥ 10 mg/L). The maximum adsorption of 64.7 mg/g for BSA was achieved as per the Langmuir adsorption model. In addition, microfiltration membrane for removal of BSA as model protein organic foulant is also studied. The effect of various MIOPs adsorbent sizes of 210, 680 and 1130 nm on the absorption capacity of BSA was investigated. Water permeability of the BSA integrated with the smallest size MIOPs membrane was increased by approximately 22% compared by the neat BSA membrane during dead-end filtration. Furthermore, the presence of small size MIOPs were also effective in increasing the permeate flux.

Investigation on the Interaction of Gabapentin with Bovine Serum Albumin by Spectroscopic Techniques

  • Ashoka, S.;Seetharamappa, J.;Kandagal, P.B.;Shaikh, S.M.T.
    • Journal of Photoscience
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    • v.12 no.3
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    • pp.113-117
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    • 2005
  • Spectroscopic investigations on the interaction of gabapentin (GBP) with bovine serum albumin (BSA) were reported. The association constant of GBP-BSA system was determined at different temperatures (298, 302, 306 and 311 K) based on the fluorescence quenching results. The GBP was found to quench the fluorescence of BSA through static mechanism. Thermodynamic parameters, the standard enthalpy change, $({\Delta}H^o)$ and the standard entropy change $({\Delta}S^o)$ were observed to be $-9.61{\pm}0.008\;kJ\;mol^{-1}$ and $3.58{\pm}0.011\;Jmol^{-1}K{-1}$ respectively. These indicated that the hydrophobic and electrostatic forces played a role in the interaction of GBP with BSA. The negative value of ${\Delta}G^o$ revealed that the binding reaction is spontaneous. The circular dichroism studies indicated the conformational changes in BSA upon interaction with GBP. The effect of some metal ions on the binding constant was also investigated.

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Effects of Boyangmaksung-bang (BYMSB) on the Membranous Nephropathy Induced by Cationic Bovine Serum Albumin in Mouse Model (보양막성방(補陽膜性方)이 Cationic Boivine Serum Albumin 투여로 유발된 Mouse의 Membranous Nephropathy에 미치는 영향)

  • Lim, Bum-Soo;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Journal of Korean Medicine
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    • v.30 no.2
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    • pp.63-78
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    • 2009
  • Objective: The purpose of experimental study was to prove the effects of Boyangmakseong-bang (BYMSB) treatment on cBSA-induced in a MN mouse model. Methods: We divided mice into 4 groups. The Normal group had no treatment. We used cBSA and induced MN mouse model to the other 3 groups. The Control group was treated with cBSA (9mg/kg i.p) only. The second group, named 'BY-250', was treated with cBSA (9mg/kg i.p) and BYMSB extract (250mg/kg, p.o). The third group, named 'BY-500', was treated with cBSA (9mg/kg i.p) and BYMSB extract (500mg/kg, p.o). After cBSA and BYMSB extract treatment for 4 weeks, the increase in percentage of body weight, proteinuria, serum albumin, total cholesterol, creatinine and BUN of all groups were measured. The CD3+, CD19+, CD4+, CD8+ cell levels of spleen of all groups were analyzed. IgA, IgG, IgM, IL-$1{\beta}$, TNF-${\alpha}$, IL-6 and IFN-${\gamma}$ levels of all groups were gauged. H&E staining, immunofluorescence staining and electron microscopy of kidney were observed. Results: BYMSB showed significant decrease in the 24hrs proteinuria, serum total cholesterol, serum IgG levels and BUN levels, and showed significant increase in the serum albumin levels compared with the control group. BYMSB showed increase in the increasing percentage of body weight and IFN-${\gamma}$ levels compared with the control. BYMSB showed decrease in the CD3+ T cells, CD4+ Th cells, IL-$1{\beta}$, TNF-${\alpha}$ and IL-6 levels, but did not show significant change compared with the control. BYMSB showed considerable decrease in the thickening of the GBM on H&E staining, deposition of IgG on immunofluorescence staining and deposition of electron-density on electron microscopy of kidney compared with the control. Conclusions: According to the above results, it is suggested that BYMSB decreases the symptoms of MN induced by cBSA in a mouse model. Therefore BYMSB seems to be applicable to MN in clinical practice.

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The Effects of Bogimakseong-bang(補氣膜性方) Treatment on cBSA-induced Membranous Nephropathy in Mouse Model (보기막성방(補氣膜性方)이 Cationized Bovine Serum Albumin투여로 유발된 Mouse의 Membranous Nephropathy에 미치는 영향)

  • Lee, Jung-Won;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Journal of Internal Korean Medicine
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    • v.29 no.4
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    • pp.1083-1099
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    • 2008
  • Objective : We aimed to identify the effects of Bogimakseong-bang(BGMSB) treatment on cBSA-induced MN in a mouse model. Methods : We divided 20 mice into 4 groups. The normal group (NR) had no treatment. We used cBSA to induced MN to the other 3 groups. One group (CT) was treated with cBSA (7mg/kg i.p) only. The second (BG-250) was treated with cBSA (7mg/kg i.p) and BGMSB extract (250mg/kg, p.o). The third group (BG-500) was treated with cBSA(7mg/kg i.p) and BGMSB extract (500mg/kg, p.o). After cBSA and BGMSB extract treatment for 4 weeks, proteinuria, serum albumin, total cholesterol, serum creatinine, BUN, total nucleated spleen cell number and total infiltrated kidney cell number of all groups were measured. CD3e+/CD19+ and CD4+/CD8 cells ratio of peripheral blood, kidney and spleen of all groups were analyzed. $IL-1{\beta}$ and $TNF-{\alpha}$, IL-6, IgG, IgM, and $IFN-{\gamma}$ levels of all groups were gauged. Histological analysis of kidney tissue and immunohistochemical staining (CD4 CD8) of kidneys were observed. Results : Proteinuria significantly decreased and serum albumin increased in groups treated with cBSA and BGMSB extract compared with the control. Total cholesterol decreased but not significantly. CD3e+/CD19cells ratio of peripheral blood decreased. CD3e+/CD19+ and CD4+/CD8 cells percentage of kidney and spleen showed no significant change. Level of $IL-1{\beta}$, $TNF-{\alpha}$ and IL-6 significantly decreased. and $IFN-{\gamma}$ increased but has not significantly. Concentration of IgG and IgM significantly decreased compared with control. Thickness of GBM decreased on histological analysis of kidney. Deposition of CD4 and CD8 decreased on immunohistochemical staining of kidney. Conclusions : According to the above result, BGMSB had a significant effect for treating MN which is cBSA-induced.

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Effects of the Acasia Catechu Extract on the Membranous Nephropathy Induced by Cationic Bovine Serum Albumin in Mice (아차(兒茶)가 Cationic Bovine Serum Albumin 투여로 유발된 Membranous Nephropathy Mouse Model에 미치는 영향)

  • Jeong, Gi-Hun;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Journal of Internal Korean Medicine
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    • v.30 no.3
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    • pp.495-509
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    • 2009
  • Objective : Membranous nephropathy(MN) is an organ-specific autoimmune disease and a relatively common cause of nephrotic syndrome in adults worldwide. But treatment of MN is not defined. This study was to evaluate the effects of Acasia Catechu extract(ACE) on the MN induced by cBSA in mice. Methods : Mice were divided into 4 groups. The normal group was injected with a saline solution. The control group was treated with cBSA(10 mg/kg i.p.) only. The third group was treated with cBSA (10 mg/kg i.p.) and ACE (250 mg/kg, p.o.). The fourth group was treated with cBSA (10mg/kg i.p.) and ACE (500mg/kg, p.o.). After cBSA and ACE treatment for 6 weeks, we measured change of body weight, 24hrs proteinuria, serum albumin, total cholesterol, triglyceride, BUN, creatinine, TNF-$\alpha$, IL-6, IL-$1{\beta}$, IFN-$\gamma$, IgA, IgM and IgG levels. The morphologic changes of renal glomeruli were also observed with a light microscope. Results : The levels of 24 hrs proteinuria, total cholesterol, triglyceride, IgG, IgM, IgA, TNF-$\alpha$, IL-6, IL-$1{\beta}$, IFN-$\gamma$ significantly decreased in both ACE groups. The level of albumin significantly increased in both ACE groups. The mRNA expression of IL-$1{\beta}$ in splenocytes considerably decreased in the ACE-500 group. In histological findings of kidney tissue, thickening of GBM decreased in both ACE groups. Conclusions : This study shows that ACE might be effective for treatment of MN. More clinical data and studies are to be done for efficient application.

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