• Title/Summary/Keyword: Bovine serum albumin

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Sealing Ability of Three Different Materials Used as Retrograde Filling

  • Park, Ji-Hoon;Kang, Seung-Bok;Choi, Yong-Hoon;Bae, Ji-Hyun
    • Journal of Korean Dental Science
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    • v.5 no.2
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    • pp.60-67
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    • 2012
  • Purpose: To test the apical leakage prevention performance of three different materials through protein leakage procedures using bovine serum albumin (BSA) and Bradford protein reagent. Materials and Methods: A total of 60 human single-rooted teeth were divided into 4 groups, and conventional root canal filling was done. The root was cut 3 mm from the apex, and a cavity was formed. Proroot MTA (MTA), Fuji II LC (GI), Fuji II LC with XP bond (GIA), and Caviton (CA) were used as experimental materials to fill the cavity in a retrograde filling manner. The extent of BSA leakage was then measured with a ultraviolet visible spectrophotometer 24, 48, and 72 hours after filling. Result: After 24 hours, among the 15 teeth of each group, 2 in MTA, 4 in GI, 3 in GIA, and 7 in CA showed leakage. After 48 hours, 3 in MTA, 5 in GI, 5 in GIA, and 10 in CA had leakage and discoloration. After 72 hours, among the 15 teeth of each group, 3 in MTA, 6 in GI, 5 in GIA, and 10 in CA showed leakage. The leakage in the CA group was greater than that in the MTA group at 48 and 72 hours based on Fisher's exact test (P=0.025), and the difference was statistically significant. Similarly, the leakage in the CA group was greater than that in the MTA group over time based on the Kaplan-Meier survival estimate (P=0.011), and the difference was statistically significant. Conclusion: Glass ionomer, glass ionomer after adhesive application, and MTA all showed leakage. Caviton showed greater leakage compared to MTA 48 and 72 hours after filling, and the difference was statistically significant; thus suggesting that Caviton is not appropriate as retrograde filling material considering its sealing ability.

AN INVESTIGATION OF IMMUNIZATION AGAINST SOMATOSTATIN BY MEASURING GROWTH AND CARCASS PARAMETERS IN GILTS

  • Du, Z.L.;Hacker, R.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.5 no.4
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    • pp.665-671
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    • 1992
  • To investigate the effects of immunization against somatostatin (SRIF) on growth rate, feed efficiency and carcass quality; forth-eight Yorkshire gilts ($age=37.5{\pm}4.3d,\;wt=8.2{\pm}1.6kg$) were randomly assigned to one of the following three treatments (1) control, (2) bovine serum albumin (BSA) and (3) SRIF. Cyclic SRIF was conjugated to BSA as the antigen containing 1 mg of SRIF diluted in 3 ml of saline. The conjugate was injected subsutaneously together with bacterial cell protein (BP) adjuvant on both sides of the neck of each gilt as the initial injection with three subsequent booster injections. Throughout the experiment all pigs were fed ad libitum a corn-soy diet containing 20% protein. Body weight and feed intake were measured on a weekly basis. All pigs in the experiment were slaughtered when they approached 101 kg body weight on the weekly weigh day. After slaughter, carcass parameters were analyzed to assess carcass quality. Results revealed that there were no differences among SRIF, BSA and control treatments for average daily gain, feed efficiency and feed intake during the first 5 wk of the experiment and from 6 wk to slaughter. The results for carcass analysis indicated that active immunization against SRIF had no effect on fat content, lean yield, water content and Canadian carcass index These data, collectively, suggest that the protocol employed in the present investigation for active immunization against SRIF is not an effective method for the enhancement of pig growth and improvement of feed efficiency and carcass quality.

Molecular Weight, Protein Binding Affinity and Methane Mitigation of Condensed Tannins from Mangosteen-peel (Garcinia mangostana L)

  • Paengkoum, P.;Phonmun, T.;Liang, J.B.;Huang, X.D.;Tan, H.Y.;Jahromi, M.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.10
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    • pp.1442-1448
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    • 2015
  • The objectives of this study were to determine the molecular weight of condensed tannins (CT) extracted from mangosteen (Garcinia mangostana L) peel, its protein binding affinity and effects on fermentation parameters including total gas, methane ($CH_4$) and volatile fatty acids (VFA) production. The average molecular weight ($M_w$) of the purified CT was 2,081 Da with a protein binding affinity of 0.69 (the amount needed to bind half the maximum bovine serum albumin). In vitro gas production declined by 0.409, 0.121, and 0.311, respectively, while CH4 production decreased by 0.211, 0.353, and 0.549, respectively, with addition of 10, 20, and 30 mg CT/500 mg dry matter (DM) compared to the control (p<0.05). The effects of CT from mangosteen-peel on in vitro DM degradability (IVDMD) and in vitro N degradability was negative and linear (p<0.01). Total VFA, concentrations of acetic, propionic, butyric and isovaleric acids decreased linearly with increasing amount of CT. The aforementioned results show that protein binding affinity of CT from mangosteen-peel is lower than those reported for Leucaena forages, however, the former has stronger negative effect on IVDMD. Therefore, the use of mangosteen-peel as protein source and $CH_4$ mitigating agent in ruminant feed requires further investigations.

Protoplast Fusion of Cellulolytic Aspergillus wentii and Aspergillus niduk (섬유소 분해효소를 생성하는 Aspergillus wentii와 Aspergillus nidulans의 원형질체 융합)

  • 성낙계;이상원;강신권;노종수;정영철
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.460-465
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    • 1990
  • Regeneration of protoplast was effective by preincubating spore suspension containing 30$\mu g$/ml of 2-DG for 4 hours, and CBE medium containing casamino acid, bovine serum albumin, ergosterol and myoinositol was found to be more efficient than any other regeneration medium tested in this experiment. The regeneration frequency was about 30%. Optimal conditions for conidial protoplast fusion were obtained by treatment of protoplasts with 10 mM $CaCl_2$ and 30% polyethylene glycol 4000 (pH 7.5) as fusogenic agent at $37^{\circ}C$ for 10 minutes. The fusion frequency was $8.2\times 10^{-4}$. The higher productivity of enzyme of fusant FWN-56 was achived: 2.3-fold for CMCase, 1.5-fold for avicelase, 1.8-fold for $\beta$-glucosidase and 2.5-fold for xylanase compared to that obtained in two parental strains. The genetic stability of fusant after maintenance on minimal medium for more than 4 weeks was high because segregant rate was below 1%. The conidial DNA content of fusant was 1.4-1.6 times higher than that of the parental strains, The nucleus size of fusants were also higher than that of each parental strains.

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N-oleoyl-D-erythro-sphingosine-based Analysis of Ceramide by High Performance Liquid Chromatography and Its Application to Determination in Diverse Biological Samples

  • Lee, Youn-Sun;Choi, Heon-Kyo;Yoo, Jae-Myung;Choi, Kyong-Mi;Lee, Yong-Moon;Oh, Sei-Kwan;Kim, Tack-Joong;Yun, Yeo-Pyo;Hong, Jin-Tae;Okino, Nozomu;Ito, Makoto;Yoo, Hwan-Soo
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.273-281
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    • 2007
  • Ceramide is involved in cell death as a lipid mediator of stress responses. In this study, we developed an improved method of ceramide quantification based on added synthetic ceramide and thin layer chromatography (TLC) separation, and applied to biological samples. Lipids were extracted from samples spiked with N-oleoyl-D-erythro-sphingosine ($C_{17}$ ceramide) as an internal standard. Ceramide was resolved by TLC, complexed with fatty-acidfree bovine serum albumin (BSA), and deacylated by ceramidase (CDase). The released sphingosine was derivatized with o-phthalaldehyde (OPA) and measured by high performance liquid chromatography (HPLC). The limit of detection for ceramide was about 1-2 pmol and the lower limit of quantification was 5 pmol. Ceramide recovery was approximately 86-93%. Ceramide concentrations were determined in biological samples including cultured cells, mouse tissues, and mouse and human plasma. TLC separation of ceramide provides HPLC chromatogram with a clean background without any interfering peaks and the enhanced solubility of ceramide by BSAceramide complex leads to the increased deacylation of ceramide. The use of an internal standard for the determination of ceramide concentration in these samples provides an accurate and reproducible analytical method, and this method can be applicable to diverse biological samples.

Kinetics of 125I-BSA Binding to Monoclonal Anti-BSA Immobilized on $0.5{\mu}m$ Polystyrene Beads (c크기의 폴리스타이렌 담체에 고정화된 단세포군 항체와 항원 BSA의 속도론)

  • ;Dan
    • KSBB Journal
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    • v.11 no.2
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    • pp.225-237
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    • 1996
  • Intrinsic binding kinetics of of 125I-bovine serum albumin (125I-BSA) and immobilized monoclonal anti-BSA (MAb 9.1) were studied. Small non-porous polystyrene beads (0.5${\mu}$m diameter) were used as a solid support to minimize the mass transfer interference on rate measurements. We demonstrated both theoretically and experimentally that the binding reaction is kinetically controlled. Rate measurements show that the association reaction is of second order and the dissociation reaction is of first order. Between 4 and $37^{\circ}C$ the measured equilibrium constant agrees well with the equilibrium constant calculated from the rate measurements. The temperature effects on association are much greater than on dissociation; the activation energy for association is about 9Kca1/mole, as compared to 2Kca1/mole for dissociation. The use of small non-porous beads as a solid support in binding studies essentially avoids mass transfer limitations; such a system makes it possible to determine the intrinsic binding characteristics of any immobilized antibody on a solid surface.

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Experimental Study in Detection of Inflammation with I-131 labeled IgG (I-131 표지 IgG를 이용한 염증 진단의 실험적 연구)

  • Kim, Deog-Yoon;Kim, Sang-Eun;Lee, Dong-Su;Ahn, Cu-Rie;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.25 no.2
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    • pp.259-265
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    • 1991
  • The purpose of this study is to investigate the ability of I-131 labeled polyclonal human immunoglobulin to localize an infection. In our country, indium-111 labeled leukocyte or Tc-99m labeled IgG are not readily available because of compex, time-consuming procedure and cost. So we tried to localize infection with I-131 labeled IgG which could be easily prepared. Six rats, infected with staphylococcus aureus in a thigh muscle, received I-131 labeled IgG intravenously and I-131 labeled bovine serum albumin (BSA) were injected to other 5 infected rats. Scintigrams were made at 1, 4, 24, 48, 72 hour later. The radiopharmaceutical demonstrated significant accumulation at the site of infection. The accumulation of I-131 labeled IgG at the site of infection was significantly (P<0.05) higher than that of I-131 labeled BSA at 48, 72 hour. Similar finding could be found at 24 hour imaging, but it was not significant statistically. Therefore it was found that vascular permeability alone could not account for the mode of action of I-131 labeled IgG and it was considered that specific binding played a role. In conclusion, focal sites of inflammation can be detected with I-131 labeled nonspecific human polyclonal IgG and it seems that this method can also be applied to localization of human infection.

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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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Antioxidative and Antimutagenic Effects and Hyperplasia Inhibitory Activity of Cancer Cells from Luffa cylindrica Extracts (수세미오이(Sponge-gourd: Luffa cylindrica L.) 추출물의 항산화, 항돌연변이 및 암세포 증식 억제 효과)

  • Oh, Se-In;Lee, Mee-Sook
    • The Korean Journal of Food And Nutrition
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    • v.25 no.4
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    • pp.888-896
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    • 2012
  • Considering the dearth of information regarding the medicinal properties of Luffa cylindrica, we assessed the antioxidative, antimutagenic and hyperplasia inhibitory activity of cancer cells from Luffa cylindrica extracts by employing biological and biochemical assays. Ethanol extracts of Luffa cylindrica inhibited MDA-BSA (malondialdehyde-bovine serum albumin) conjugation reaction ($66.38{\pm}2.65$), DPPH (1,1-diphenyl-2-picryl-hydrazyl) radical production ($60.13{\pm}0.42$) and lipid peroxidation ($56.04{\pm}3.24$). In this study, Luffa cylindrica is believed to exert possible antioxidative effects. The direct and indirect antimutagenic effects of the ethanol extracts of Luffa cylindrica were examined by the Ames test using Salmonella typimurium TA98 and TA100. The inhibitory effects on indirect and direct mutagenicity shows an weak tendency, particularly in direct mutagenicity mediated by 2-nitrofluorene in Salmonella typimurium TA98 ($5.82{\pm}5.74$) and in indirect mutagenicity mediated by 2-anthramine in Salmonella typimurium TA100 ($5.76{\pm}2.15$). The ethanol extracts of Luffa cylindrica on cancer cell hyperplasia inhibitory activity via MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay exerted cytotoxic effects on Hela cells ($55.83{\pm}3.83$) and MCF-7 cells ($33.03{\pm}2.09$), which were used in this study. Based on these results, it believed that the ethanol extracts of Luffa cylindrica have antioxidative capacities as well as hyperplasia inhibitory activity of cancer cells. Furthemore, Luffa cylindrica is a candidate for the prevention and dietetic treatment of chronic diseases and for the development of functional food.

Immobilization of Trypsin onto Silk Fibroin Fiber via Spacer Arms

  • Lee, Ki-Hoon;Kang, Gyung-Don;Shin, Bong-Seob;Park, Young-Hwan;Nahm, Joong-Hee
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.195-200
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    • 2004
  • Trypsin can be immobilized on silk fibroin fiber (SFF) by introducing several spacer arms, such as ethylene diamine (ED), bovine serum albumin (BSA) and silk sericin (SS). Direct immobilization on silk fiber (SFFGA) has low activity because of the steric hindrance between the trypsin and substrate. The introduction of spacer arms onto SFF-GA can enhance the activity of trypsin by reducing the steric hindrance. When ED is used as a spacer arm, the activity of trypsin has increased but its stability decreased due to the increased hydrophobicity of SFF. BSA and SS, as a spacer arm, have better results in both activity and stability. SFF-BSA shows some decrease in the specific activity due to improper immobilizatin. SFF-SS maintained 90% of its initial activity even after 12 hrs incubation at $50^{\circ}C$. In the case of repeated hydrolysis of silk sericin with immobilized trypsin, SFF-GA and SFF-ED lost 50% of their initial activity right after first run, whereas SFF-BSA and SFF-SS maintained 80% of their initial activities even after 5 runs. Higher operational stability is due to increased hydrophilicity of SFF by introducing hydrophilic spacer arms such as BSA and SS. The high content of serine in SS increases the hydrophilicity of SFF resulting the best results among other spacer arms.