• 대한수의학회지
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• 제5권1호
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• pp.1-16
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• 1965

I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

• 대한한방내과학회지
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• 제30권3호
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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.

• 동의생리병리학회지
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• 제23권5호
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• pp.1063-1072
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• 2009

Membranous nephropathy(MN) is the most common cause of adult nephrotic syndrome worldwide. But treatment of MN is not defined. This study was to evaluate the effects of Lonicerae Flos Extract(LFE) on the MN induced by cBSA in mice. Mice were divided into 4 groups. The first group named for 'Normal' was injected with a saline solution. The second group named 'Control' treated with cBSA(10 mg/kg i.p) only. The third group named 'LFE-250', treated with cBSA(10 mg/kg i.p) and LFE(250 mg/kg, p.o). The fourth group named 'LFE-500'treated with cBSA(10 mg/kg i.p) and LFE(500 mg/kg, p.o). After cBSA and LFE treatment for 4 weeks, we measured change of body weight, 24hrs proteinuria, serum albumin, total cholesterol, triglyceride, BUN, creatinine, TNF-$\alpha$, IL-6, IL-$1{\beta}$, IL-10, IFN-$\gamma$, IgA, IgM and IgG levels. The morphologic changes of renal glomeruli were also observed with a light microscope. The levels of 24 hrs proteinuria, total cholesterol, IgG , IgM, IgA, IL-6 were significantly decreased in both LFE groups. The level of triglyceride, IL-$1{\beta}$ was significantly decreased in LFE-500 group. The level of Albumin was significantly increased in LFE-250 group. The level of TNF-$\alpha$, IFN-$\gamma$ were significantly decreased in LFE-250 group. The mRNA expression of IL-$1{\beta}$ in splenocytes was consideraly decreased in LFE-500 group. In histological findings of kidney tissue, thickening of GBM decreased in both LFE groups. This study shows that the LFE might be effective for treatment of MN. More clinical data and studies are to be done for efficient application.

• Journal of Photoscience
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• 제11권1호
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• pp.29-33
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• 2004

The mechanism of interaction of cyanocobalamin (CB) with bovine serum albumin (BSA) has been investigated by spectrofluorometric and circular dichroism methods. Association constant for the CB-BSA system showed that the interaction is non-covalent in nature. Binding studies in the presence of an hydrophobic probe, 8-anilino-l-naphthalene sulphonic acid, sodium salt (ANS) showed that there is hydrophobic interaction between CB and ANS and they do not share common sites in BSA. Stern-Volmer analysis of fluorescence quenching data showed that the fraction of fluorophore (protein) accessible to the quencher (CB) was close to unity indicating thereby that both tryptophan residues of BSA are involved in drug-protein interaction. The rate constant for quenching, greater than $10^{10}$ $M^{-1}$ $s^{-1}$, indicated that the drug binding site is in close proximity to tryptophan residue of BSA. Thermodynamic parameters obtained from data at different temperatures showed that the binding of CB to BSA involves hydrophobic bonds predominantly. Significant increase in concentration of free drug was observed for CB in presence of paracetamol. Circular dichroism studies revealed the change in helicity of BSA due to binding of CB to BSA.

• Bulletin of the Korean Chemical Society
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• 제32권6호
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• pp.2063-2069
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• 2011

We studied the interaction of 3,3',3'',3'''-ethylenetetrakis-4-hydroxycoumarin (EHC) with bovine serum albumin (BSA) in acetate buffer and phosphate buffer with different pH values by UV-vis absorption spectrometry and fluorescence spectrometry respectively. It was found that the pH values of the buffer solutions had an effect on the interaction process. In acetate buffer of pH 4.70, the carbonyl groups in EHC bound to the amino groups in BSA by means of hydrogen bond and van der Waals force, which made the extent of peptide chain in BSA changed. By contrast, in phosphate buffer of pH 7.40, hydrophobic force played a major role in the interaction between EHC and BSA, while the hydrogen bond and van der Waals force were also involved in the interaction. The results of spectrometry indicated that BSA could enhance the fluorescence intensity of EHC by forming a 1:1 EHC-BSA fluorescent complex through static mechanism at pH 4.70 and 7.40 respectively. Furthermore, EHC bound on site 1 in BSA.

• Bulletin of the Korean Chemical Society
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• 제32권9호
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• pp.3261-3266
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• 2011

The interaction of bilobalide (BB) and ginkgolides B (GB) with bovine serum albumin (BSA) was investigated by fluorescent technique and UV/vis absorption spectroscopy. The results showed that BB and GB could intensively quench the fluorescence of BSA through a static quenching procedure. The binding constants (Ka) and the average binding distance between the donor (BSA) and the acceptor (ginkgolides) were obtained ($r_{BB}$ = 5.33 nm and $r_{GB}$ = 4.20 nm) by the theory of non-radiation energy transfer, and then the thermodynamic parameters such as ${\Delta}S^0$ (0.17-0.32 kJ/mol), ${\Delta}G^0$ (-20.76 ~ -17.79 kJ/mol) and ${\Delta}H^0$ (32.47-76.52 kJ/mol) could be calculated, respectively. All these results revealed that the interaction of BB and GB with BSA were driven mainly by hydrophobie force. The synchronous fluorescence spectroscopy was applied to examine the effect of two ginkgolides on the configuration of BSA. The configuration alteration of BSA could be induced by the hydrophobicitv environment of tyrosine with the increase of the drug concentration.

• 대한약침학회지
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• 제14권3호
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• pp.71-79
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• 2011

Objectives : This study was undertaken to identify fibrinolytic proteases from Gloydius blomhoffii siniticus venom and to characterize a major fibrinolytic protease purified from the venom. Methods : The venom was subjected to chromatography using columns of Q-Sepharose and Sephadex G-75. The molecular weights of fibrinolytic proteases showing fibrinolytic zone in fibrin plate assay were determined in SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) The effects of inhibitors and metal ions on fibrinolytic protease and the proteolysis patterns of fibrinogen, gelatin, and bovine serum albumin were investigated. Results : 1) The fibrinolytic fractions of the three peaks isolated from Gloydius blomhoffii siniticus venom contained two polypeptides of 46 and 59 kDa and three polypeptides of 32, 18, and 15 kDa and a major polypeptide of 54 kDa, respectively. 2) The fibrinolytic activity of the purified protease of 54 kDA was inhibited by metal chelators, such as EDTA, EGTA, and 1,10-phenanthroline, and disulfhydryl-reducing compounds, such as dithiothreitol and cysteine. 3) Calcium chloride promoted the fibrinolytic activity of the protease, but mercuric chloride and cobalt(II) chloride inhibited it. 4) The fibrinolytic protease cleaved preferentially A${\alpha}$-chain and slowly B${\beta}$-chain of fibrinogen. It also hydrolyzed gelatin but not bovine serum albumin. Conclusions : The Gloydius blomhoffii siniticus venom contained more than three fibrinolytic proteases. The major fibrinolytic protease was a metalloprotease which hydrolyzed both fibrinogen and gelatin, but not bovine serum albumin.

• 약학회지
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• 제33권5호
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• pp.308-311
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• 1989

Binding parameters of some edible dyes with bovine serum albumin were determined at pH 7.4 buffer solution. The edible dyes used were fast green, brilliant blue FCF, indigo carmine and allura red AC, and the final concentrations of each dyes were $3{\sim}9{\times}10^{-6}M$, $6{\times}10^{-6}M$, $3{\times}10^{-5}M$ and $3{\times}10^{-5}M$, respectively. The final concentrations of bovine serum albumin were $1{\sim}7{\times}10^{-5}M$. The values of binding free energy between edible dye and protein were ranged from -6,024 to -6,800 cal/mole.

• 센서학회지
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• 제30권5호
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• pp.326-330
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• 2021

Reactions between gold (Au) surface plasmon resonance (SPR) chips and bovine serum albumin (BSA) dissolved in solutions of different pH were investigated. The charge on the BSA depends on the pH of the solution in which it is dissolved. Thus, dissolving BSA in different pH solutions resulted in different charges of BSA. Among the BSA dissolved in solutions with pH 4.01, 7.4, and 10.01, the SPR response was the highest for BSA dissolved in the solution of pH 4.01. To eliminate the response variation owing to the difference in the refractive indices of the solutions, phosphate buffered saline (PBS) was injected into the system after the reaction of BSA with the Au SPR chip had happened. In this case too, the BSA dissolved in the solution with pH 4.01 exhibited the highest response. This may be attributed to the non-uniform distribution of ionic patches on the BSA, which can induce electrostatic attraction to the surface even though BSA has a positive charge at pH 4.01, and the absolute values of the net charge of BSA at pH 4.01 and 7.4 were very close.

• 대한수의학회지
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• 제38권2호
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• pp.394-400
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• 1998

Cryopreservation of embryos by vitrification is a simple method to preserve bovine embryos for subsequent embryo transfer, but embryonic viability after vitrification has been inconsistent and low compared with conventional slow freezing. The aim of the present study is to examine the effect of serum or serum albumin in a vitrification solution and epidermal growth factor(EGF) or fibroblast growth factor(FGF) on in vitro viability of bovine blastocysts frozen by vitrification. Bovine blastocysts were produced by in vitro maturation, fertilization of follicular oocytes and culture of embryos in a synthetic oviduct fluid medium(SOFM) containing BSA and 19 essential and nonessential amino acids. Blastocysts with excellent or good morphology were selected at 7 or 8 days after culture and utilized for vitrification. In experiment 1, blastocysts were vitrified in a solution containing semi-fetal calf serum(SFCS) or BSA(5 or 10mg/ml) and then their subsequent viabilities were examined by culturing thawed embryos in a SOFM containing BSA and 19 amino acids. Effect of EGF or FGF added to a SOFM containing polyvinyl alcohol(PVA) on the viability of vitrified-thawed blastocysts was investigated in experiment 2. BSA added at 5 or 10mg/ml to a vitrification solution showed significantly higher(p < 0.05) developmental rate to expanded and hatching blastocysts than SFCS, but there was no significant difference in the developmental rate to hatched blastocysts after thawing. Supplementation of a culture medium with EGF and/or FGF significantly increased(p < 0.05) embryo development to expanded blastocysts compared with control but showed no beneficial effect on the development to hatching or hatched blastocysts. Coculture of thawed embryos with granulosa cells in a TCM 199 containing 10% fetal calf serum(FCS) showed the highest developmental rate to expanded, hatching and hatched blastocysts among the groups tested. In conclusion, supplementation of a vitrification solution with BSA at 5mg/ml and culture of thawed blastocysts in a medium containing EGF and/or FGF can improve in vitro viability of bovine blastocysts frozen by vitrification.