• 한국산학기술학회논문지
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• 제19권6호
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• pp.147-153
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• 2018

이중유화법을 통하여 BSA 수용액을 봉입하는 PLGA 마이크로캡슐 입자를 제조하였다. 마이크로캡슐 입자 외부의 수용액 상에서는 입자 표면의 경화를 위하여 유화제를 금속염으로 대체하였다. 제조한 마이크로캡슐 입자에 대하여서 모폴로지, 입자직경 BSA 봉입효율 및 in-vitro 방출실험을 수행하였다. 전자현미경(SEM) 촬영을 통하여 마이크로캡슐 입자는 매끄러운 표면의 특성을 지녔으며 $1-7{\mu}m$ 범위의 직경을 갖는 것이 확인되었다. 마이크로캡슐 입자의 모폴로지는 주로 일차 유화액 내 내부 수용액 대비 고분자 용액의 부피 비 그리고 입자 외부 수용액에서의 금속염의 농도에 직접적인 영향을 받았다. 또한 이러한 요소는 단백질 봉입효율과 in-vitro 방출에도 일부 영향을 미쳤다. 본 실험에서 제조한 마이크로캡슐 입자는 in-vitro 방출 실험에서 초기에 높은 유속의 방출 현상을 보였다. 그렇지만 본 연구에서 제조한 마이크로캡슐 입자는 다른 연구의 결과에 비해 긴 기간 동안의 방출을 보였다. 이상의 결과를 통해 2가의 금속염이 마이크로캡슐 입자의 제조에서 유화제를 대체할 수 있는 좋은 방편이 될 수 있다는 결론을 내릴 수 있었다.

• 생약학회지
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• 제39권3호
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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.

• Journal of Pharmaceutical Investigation
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• 제19권3호
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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.

• KSBB Journal
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• 제22권1호
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• pp.48-52
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• 2007

본 연구에서는 sonificator를 장착하여 세포막을 파쇄하고 현장에서 형광을 이용하여 조작이 간편하고 단시간에 DNA와 단백질을 동시에 측정할 수 있는 자동화된 형광기를 개발하기 위하여 단백질을 측정하는데 최적의 파쇄조건을 확립하고자 하였다. 용액 중에 녹아 있는 공기 중의 oxygen은 collisional quenching을 일으키는데 sonification이나 가열처리를 시키면 oxygen이 제거되어 quenching 효과가 크게 감소되어 높은 형광값을 나타내었다. 0.7 X 이상의 형광시약 농도에서는 반응시작 후 1분 이내에 측정해야 하며, 0.3 X 이하의 형광시약 농도에서는 반응시작 후 2$\sim$3분 사이에 반응을 시킨 후 측정하는 것이 바람직한 것으로 나타났다. $100{\mu}g/m{\ell}$ 이상의 BSA 농도에서는 형광시약이 포화되었으며, 시료를 sonification시키면 단백질이 변성되어 눈에 보일 정도로 불투명해져서 시료 용액의 불투명도로 인해 형광 값이 감소되는 경향을 나타내었으며, $1{\mu}g/m{\ell}$ 이하의 BSA 시료에서는 sonification을 시키지 않은 시료보다 sonification을 시켰을 때 $0.125{\mu}g/m{\ell}$의 BSA를 훨씬 더 구분이 잘 되어 낮은 단백질 농도에서는 sonification시키는 것이 훨씬 유리한 것으로 나타났다.

• 한국미생물·생명공학회지
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• 제28권1호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제15권3호
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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.

• 한국가축번식학회지
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• 제15권2호
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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%).

• 한국식품위생안전성학회지
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• 제10권4호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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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%).

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권3호
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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.