• Title/Summary/Keyword: G-BSA

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Effects of process parameters on encapsulations of BSA aqueous solutions into PLGA microcapsule particles using double emulsion technique (이중유화법을 통하여 BSA 수용액을 PLGA 마이크로캡슐 입자에 봉입하는 과정에서의 공정변수의 영향)

  • Kwon, Sejin;Koo, Ja-Kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.6
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    • pp.147-153
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    • 2018
  • PLGA microcapsule particles encapsulating BSA aqueous solutions were prepared using a water-in-oil-in-water emulsion method. The morphology, particle size, BSA encapsulating efficiency, and in-vitro release test were also studied using the microcapsule particles. In the outer aqueous phase, an emulsifier, e.g., PVA, was replaced with metal salts for surface solidification. Scanning electron microscopy (SEM) showed that the microcapsule particles had smooth surfaces and were between $1{\mu}m$ and $7{\mu}m$ in size. The microcapsule particle morphology was affected directly by the ratio between the polymer solution and inner aqueous solution, and composition of the outer aqueous solution. The factors also partially affected the BSA encapsulation efficiencies and in-vitro release rates. All the microcapsule particles showed an initial burst release through the in-vitro release test. On the other hand, the particles also showed a relatively long release period. Metal salts could be good choices to replace the emulsifier to solidify the microcapsule particle surfaces.

Inhibitory Effects of the Seeds of Cornus officinalis on AGEs Formation and AGEs-induced Protein Cross-linking (산수유 씨의 최종당화산물의 형성 및 교차결합에 미치는 효과)

  • Kim, Chan-Sik;Jang, Dae-Sik;Kim, Jung-Hyun;Lee, Ga-Young;Lee, Yun-Mi;Kim, Young-Sook;Kim, Jin-Sook
    • Korean Journal of Pharmacognosy
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    • v.39 no.3
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    • pp.249-254
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    • 2008
  • An 80% EtOH extract and the solvent fractions of the seeds of Cornus officinalis were evaluated for their inhibitory activities against advanced glycation end products (AGEs) formation and AGEs-induced protein cross-linking in vitro. In vitro assay for AGEs-bovine serum albumin (BSA) formation showed that the 80% EtOH extract, n-hexane, EtOAc, n-BuOH and water fractions significantly inhibited AGEs formation with observed $IC_{50}$ values of 1.13, 17.64, 1.52, 1.24 and $3.27{\mu}g/ml$, respectively. In indirect AGEs-ELISA assay, the 800% EtOH extract, EtOAc and n-BuOH fractions exhibited more potent inhibitory activity on AGEs-BSA formation than aminoguanidine, a well know AGEs inhibitor. Furthermore, the 80% EtOH extract and all the solvent fractions inhibited concentration-dependently AGE-BSA cross-linking to collagen. The 80% EtOH extract, EtOAc, n-BuOH and water fractions also had a breaking activity against preformed AGE-BSA cross-linking concentration dependently. Thus these results suggest that the 80% EtOH extract and fractions of the seeds of C. officinalis could be an inhibitor as well as breaker of AGE-BSA cross-linking.

Drug-Biomacromolecule Interaction (XIII)-Effect of ionic Strength, pH and Temperature on Binding of Cephalothin to Bovine Serum Albumin- (약물과 생체고분자 간의 상호작용(제 13보)-세파로친과 소혈청알부민의 결합에 미치는 이온강도, pH 및 온도의 영향)

  • Kim, Chong-Kook;Lim, Yun-Su;Yang, Ji-Sun;Jeong, Eun-Ju
    • Journal of Pharmaceutical Investigation
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    • v.19 no.3
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    • pp.163-171
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    • 1989
  • To investigate the protein binding characteristics of cephalothin, the effects of ionic strength, pH and temperature on the binding of cephalothin to bovine serum albumin (BSA) were studied by UV difference spectrophotometric method. With increasing ionic strength at constant PH and temperature, association constant decreased, but the number of binding sites sites was about 2 constantly. It may be deduced that the binding process is not only due to electrostatic forces. And the increased association constant at high ionic strength is explained by conformational changes of BSA from complex to subunits. The pH effect on the affinity of interaction indicated that the binding affinity of drug is higher in the neutral region than in the alkaline region. And, at high pH value, the number of binding sites decreased from 2 to 1 because of the conformational changes of BSA in alkaline region. The decrease in binding affinity of BSA to drug with increasing temperature was characteristic of an exothermic reaction. And the negative sign of ${\Delta}G^{\circ}$ meant that the binding process occurs spontaneously under the experimental conditions. In cephalothin-BSA complex formation, since the net enthalpy change value and entropy change value are positive, it is assumed that hydrophobic bindings are predominant in this binding process.

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The Early Detection of the Protein Toxin using Sanification and Fluorescent Dye in the Field (현장에서 초음파 파쇄와 형광시약을 이용한 단백질 독소의 조기 탐지)

  • Ha, Yeon-Chul;Choi, Ki-Bong;Kim, Seong-Joo;Choi, Jung-Do
    • KSBB Journal
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    • v.22 no.1
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    • pp.48-52
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    • 2007
  • This study was carried out to establish the optimum disruption condition of a sonificator for the protein toxin for the purpose of developing automatic biological agent detector equipped a sonificator. One of the best-known collisional quenchers is molecular oxygen, which quenches almost all known fluorophores. The sonification does an excellent job of degassing, which decreased the quenching effect and increased the fluorescence quantity. The fluorescence measurement for the protein using 0.7 X fluorescent dye concentration and above must be done in 1 minute and the fluorescence measurement for the protein using 0.3 X fluorescent dye concentration and below has to be done between 2 and 3 minute. The fluorescence quantity of the sonificatied protein sample was much higher that of the non-sonificatied protein sample. Sonificating the sample turned out to be favorable for the fluorescence measurement when measuring at the low protein concentration.

Probiotic Effect of Lactobacillus reutri BSA-131 on Piglets (자돈에 투여한 Lactobacillus reutri BSA-131의 생균제 효과)

  • 장영효;김종근;김홍중;김원용;김영배;박용하
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.8-13
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    • 2000
  • A study was carried out to determine the probiotic effect of Lactobacillus reuteri BSA-131 by investigating the growth performance and fecal microbial population of piglets. Five dietary treatment groups, the basal diet (control, BD), basal diet with antibiotics(BA), basal diet with 2$\times$106/g of probiotics (BP6), 2$\times$108/g of probiotics (BP8) and basal diet with antibiotics and 2$\times$108/g probiot-ics(BAP8) were divised. Each dietary treatment group was consisted of 1 month of age piglets(male 13, female 12). Fecal micro-flora, body weights and feed consumption were measured at before, after and stop feeding of probiotics. The results showed that the CFU of fecal Enterobacteriaceae of piglets of the group BA, BP6, BP8 and BAP8, were reduced (P<0.05) compared to control BA. On the contrary, Lactobacillus counts were increased significan시 (P<0.001) in all groups fed probiotics dites, but not antibiotics. Body weight of probiotics treated piglets were improved 5% (p<0.001) in BP6 group than that of control group and antibiotic treated piglets BAP group was 27% (P<0.001) higher than BA group. The amount of feed consumption value of probiotics treated piglets showed 21-30% (P<0.001) lower intake than the control group, whereas antibiotic treated piglets BAP was 20% (P<0.001) higher than BA group. The results showed that body weights and feed to gain ratios were improves 19% when compared to control piglets for groups fed diets probiotic. It is very suggestive that productivity of probiotic piglets would be economical in pig farming.

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Secretory Proteins from Goat Oocytes Matured in Culture

  • Malakar, Dhruba;Majumdar, A.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.3
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    • pp.340-345
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    • 2002
  • In this experiment, oocytes were collected from goat ovaries available in slaughterhouse by follicle puncture method. Morphologically culturable type of oocytes which having compact, multilayered cumulus granulosa cell complex and evenly granulated cytoplasm, was separated under a stereozoom microscope. Oocytes were washed thoroughly in maturation medium containing TCM-199, $1{\mu}g/ml$ estradiol-$17{\beta}$, 0.5 ${\mu}g/ml$ FSH, $100{\mu}g/ml$ LH, 3 mg/ml BSA and 10% estrus goat serum. Washed oocytes were cultured into maturation medium on granulosa cell monolayer. Culture plate was then kept into $CO_2$ incubator at $38{\pm}1^{\circ}C$, maximum humidity and 5% $CO_2$ for 18 h. After maturation the oocytes were washed thoroughly with maturation medium containing polyvinyl alcohol (PVA) without serum and BSA and further cultured for 12 h for secretory proteins of oocytes. PVA medium was collected, pooled and concentrated by 5000 cut off centrisart. Secretory proteins were separated on 12.5% SDS-PAGE. A total number of 3.41 oocytes per ovary were obtained and 2.17 culturable oocytes per ovary were cultured into maturation medium. After 18 h of maturation, 4,567 oocytes (1.82 oocytes per ovary) were further cultured into serum and BSA free PVA medium for its secretory proteins. Four secretory proteins of oocytes with approximately molecular weight of 45, 55, 65 and 95 kDa were obtained on SDS-PAGE in silver staining and three proteins with approximately molecular weight of 45, 55 and 65 kDa in Coomassie brilliant blue staining. In conclusion, four secretory proteins with approximately molecular weight of 45, 55, 65 and 95 kDa was obtained from in vitro cultured oocytes of goats.

Studies on the Effects of the Capacitation Methods of Spermatozoas on in-vitro Fertilization and Cleavage Rate of Bovine Follicular Oocytes (수정능획득 처리법이 소 난포란의 체외수정 및 분할율에 미치는 영향에 관한 연구)

  • 김상근;한성욱;한방근
    • Korean Journal of Animal Reproduction
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    • v.15 no.2
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    • pp.125-132
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    • 1991
  • The studies on the carried out to investigate the effects of capacitation method of spermatozoa on the in vitro fertilization and cleavage rate of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible of diameter 3~5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and FCS for 24~48hrs in an incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18hrs with motile capacitated sperm by preincubation of mKRB, treatment of HIS(high strength ion), Ca-IA(Inophore A), BFF(bovine follicular fluids) and heparin. The results obtained in these experiments were summarized as follows : 1. The in vitro fertilizatin and cleavage rate offollicular oocytes fertilized with capacitated spermatozoas in BO solution by preincubation of mKRB, treatment of HIS, Ca-IA, BFF and heparin method were 53.1%, 33.9%, 50.8%, 48.1%, 58.8% and 28.1%, 17.7%, 26.2%, 22.8%, 32.8%, respectively. And the fertilization and cleavage rate of heparin method was of highest of all. 2. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solutin by both caffeine, BSA and heparin methods were 65.8%, 70.3% and 40.8%, 47.3%, respectively, and those rates were higher treatment of heparin+BSA, heparin+caffeine than treatment of heparin. 3. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoa in BO solution with heparin concentrations of 2, 5, 10, 20, 40$\mu\textrm{g}$/ml were 50.0%, 54.7%, 58.1%, 51.7% and 27.9%, 32.8%, 37.1%, 30.0%, respectively. And the fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution with 10$\mu\textrm{g}$/ml of heparin was the highest of all. 4. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with caffeine concentraton of 10, 20, 30, 40$\mu\textrm{g}$/ml were 71.4%, 74.3% and 70.6%, 70.0% and 45.7%, 47.3%, 44.1%, 41.4%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with caffeine and heparin together(70.3~74.3%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%). 5. The in vitro fertilization and cleavage rate of follicular oocytes fertilized with capacitated spermatozoas in BO solution containing heparin with BSA concentration of 5, 10, 20, 30$\mu\textrm{g}$/ml were 63.6%, 62.9%, 66.7%, 60.3% and 44.1%, 43.5%, 48.5%, 42.7%, respectively. The fertilization and cleavage rate of spermatozoa fertilized in BO solution with BSA and heparin together(60.3~66.73%) was higher than that of spermatozoa fertilized in BO solution with heparin(58.8%).

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Study on the Accumulation of Ochrtoxin A in Mouse's Organs and the Establishment of ELISA Method for Ochratoxin A (Ochratoxin A가 마우스의 장기에 축적 정도와 ELISA법 확립에 관한 연구)

  • 김동술
    • Journal of Food Hygiene and Safety
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    • v.10 no.4
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    • pp.225.2-262
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    • 1995
  • Ochratoxin A was produced from Aspergillus ochraceus ATCC 18472 which was then orally administered into the experimental mice to study the toxic levels of ochratoxin A. AELISA (enzyme-linked immunosorbent assay) method which is more rapid and safe than conventional analytical method, was developed by using ochratoxin A antibody. This method was successfully used to measure the levels of ochratoxin Ain blood, liver and kidney of mice. In order to produce a large amount of ochratoxin A to study toxicity in the mice, Aspergillus ochraceus ATCC 18412 was incubated in the rice medium and as a result 0.5 g of ochratoxin A form rice medium (kg) was produced after extraction and purification Feed consumption and gain in body weight of mice with ochratoxin A at a level of $10 \mu\textrm{g}/g$ body weight was significantly (p<0.05) reduced as compared with control during period of 3 weeks. Ochratoxin A-BSA conjugate was made by putting 13 mole of ochratoxin A on I mole of BSA. This conjugate was used to develop ELISA method. The minmum detection level of ochratoxin A by established ELISA method was 0.5 ppb. After oral adminstraton of ochratoxin A dose of $10\;\mu\textrm{g}/g$ every two day for 3 weeks, concentration of ochratoxin A was measured in the blood, liver and kidney by ELISA method. The level of ochratoxin A was $11\;\mu\textrm{g}/dl,\;0.9 \mu\textrm{g}/g\;and\;3.7\;\mu\textrm{g}/g$ in the blood, liver and kidney, respectively.

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Effects of Addition of Pyruvate, Lactate, Calcium, and Protein Sources on the Development of Bovine IVF Embryos

  • Lee, S.H.;Lee, J.H.;Chung, G.M.;Im, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.6
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    • pp.655-660
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    • 1998
  • To produce blastocysts more efficiently, it is required to identity accurately the factors involving embryonic cleavage in the chemically defined medium. Effects of pyruvate, lactate, calcium and protein sources on early cleavage of bovine follicular oocytes were investigated. The percentage of IVF embryos cleaved to ${\geq}$ 2-cell or ${\geq}$ 8-cell was higher in pyruvate (+) and lactate (+) (48 or 14%) than in pyruvate (-) and lactate (-) (22% or 4%), than in pyruvate (+) and lactate (-) (28% or 5%) and than in pyruvate (-) and lactate (+) (40% or 10%). Lactate was more effective than pyruvate during early cleavage of bovine embryos in the chemically defined medium. The percentage of IVF embryo cleaved to ${\geq}$ 2-cell and ${\geq}$ 8-cell in calcium (-) (19 and 6%) was significantly (p < 0.05) lower than in calcium (+) (78 and 45%). The percentage of embryos developed to ${\geq}$ 2-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (57, 57 and 57%). Also the percentage of A grade embryos developed to ${\geq}$ 2-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (40, 35 and 28%). The percentage of embryos developed to ${\geq}$ 8-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (33, 23, and 22%). However, the percentage of A grade embryos developed to ${\geq}$ 8-cell in BSA (24%) was significantly (p < 0.05) higher than in 1 and 20% FBS (13 and 8%). The percentage of embryos developed to ${\geq}$ morula showed no significant (p < 0.05) difference among BSA, 1, 10 and 20% FBS (76, 76, 80 and 68%). The percentage of A grade embryos developed to ${\geq}$ morula in 10% FBS (59%) was significantly (p < 0.05) higher than 20% FBS (43%). The percentage of embryos developed to blastocyst showed no significant (p < 0.05) difference among BSA, 1, 10 and 20% FBS (34, 41, 43 and 32%). However, the percentage of A grade embryos developed to ${\geq}$ blastocysts in 10% FBS (25%) was significantly (p < 0.05) higher than in 20% FBS (8%).

The Influence of Bakers' Yeast Cells on Protein Adsorption in Anion Exchange Expanded Bed Chromatography

  • Mei Chow Yen;Ti Tey Beng;Ibrahim Mohammad Nordin;Ariff Arbakariya;Chuan Ling Tau
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.280-283
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    • 2005
  • The adsorption of a model protein bovine serum albumin (BSA) in expanded bed chromatography was undertaken by exploiting a commercially available expanded bed column (20 mm i.d.) from UpFront Chromatography and Streamline DEAE $(\rho=1.2g/cm^3)$ from Amersham Pharmacia Biotechnology. The influence of whole yeast cells on the adsorption capacity of column was explored by employing yeast cells in a concentration ranged of 0 to $15\%(w/v)$. Equilibrium isotherms for adsorption of BSA on Streamline DEAE were correlated by using Langmuir equation. The presence of yeast cells resulted in decreased of BSA binding capacity in both batch binding and expanded bed chromatography. Results indicated that the yeast cells act as competitor for proteins to bind to the sites on adsorbents.