• Title/Summary/Keyword: buffer stabilization

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Impact of Solvent pH on Direct Immobilization of Lysosome-Related Cell Organelle Extracts on TiO2 for Melanin Treatment

  • Bang, Seung Hyuck;Kim, Pil;Oh, Suk-Jung;Kim, Yang-Hoon;Min, Jiho
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.718-722
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    • 2015
  • Techniques for immobilizing effective enzymes on nanoparticles for stabilization of the activity of free enzymes have been developing as a pharmaceutical field. In this study, we examined the effect of three different pH conditions of phosphate buffer, as a dissolving solvent for lysosomal enzymes, on the direct immobilization of lysosomal enzymes extracted from Hen's egg white and Saccharomyces cerevisiae. Titanium(IV) oxide (TiO2) nanoparticles, which are extensively used in many research fields, were used in this study. The lysosomal enzymes immobilized on TiO2 under each pH condition were evaluated to maintain the specific activity of lysosomal enzymes, so that we can determine the degree of melanin treatment in lysosomal enzymes immobilized on TiO2. We found that the immobilization efficiency and melanin treatment activity in both lysosomal enzymes extracted from Hen's egg white and S. cerevisiae were the highest in an acidic condition of phosphate buffer (pH 4). However, the immobilization efficiency and melanin treatment activity were inversely proportional to the increase in pH under alkaline conditions. In addition, enhanced immobilization efficiency was shown in TiO2 pretreated with a divalent, positively charged ion, Ca2+, and the melanin treatment activity of immobilized lysosomal enzymes on TiO2 pretreated with Ca2+ was also increased. Therefore, this result suggests that the immobilization efficiency and melanin treatment activity of lysosomal enzymes can be enhanced according to the pH conditions of the dissolving solvent.

A Load Distribution System on P2P Online Game Based on RS Reconfiguration by Interesting Regions (P2P 온라인 게임에서의 관심영역별 영역관리자 재구성 기반 부하분산 시스템)

  • Jung, Mi-Sook;Kim, Seong-Hoo;Park, Kyoo-Seok
    • Journal of Korea Multimedia Society
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    • v.12 no.3
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    • pp.345-353
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    • 2009
  • It is for sure that a stable game managing system is absolutely needed to accept simultaneous interfacing of many users for P2P on-line game system. The proposed P2P on-line game system in this paper is able to get smart and stable game managing taking care of extensive players through reorganizing numerous RS(Region Server) and mutual communications among RS's which can be avoid congestion on one region. Moreover, it is possible to synchronize for game nodes in time stamp utilizing Global Zone Buffer of Monitoring Server which leads to breakup loads. The system manages middleware layers in the so-called sub area, and it is able to execute no matter how big the game sizes are. That means, in some ways, we got everything we try to ensure such as the problems of high cost server and stabilization of message transmission. This paper welcomes to prove efficiency of the suggested system through the simulation.

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A Study on Development of Boar Semen Extender Kp for Swine AI I. Stabilization of pH Change and In Vitro Survival of Frozen-Thawed Boar Sperm in Kp Extender (돼지인공수정용 정액액상보존제 Kp 의 개발에 관한 연구 I. Kp 의 pH 조절과 냉동정자에 의한 보존성 검정)

  • 김선의;정구민;서동삼;김득중;김인철;김현종;신영수;임경순
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.405-410
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    • 1998
  • Boar semen extender Kp (Hankook Life-Science, Korea) was newly formulated by authors. This study was carried out to investigate the optimal concentrations of EDTA, Tris and citrate buffers in the Kp extender (Basic Kp) on the pH change during storage. And then the motility of boar sperm with the Kp pH stabilized (Modified Kp) was compared with those of commercial products imported into Korea such as BTS (Mini-tube. Germany; BTSg), BTS (Tri-bio, USA; BTSa) and Modena (SGI, USA). The pH values of all extenders were increased gradually with the storage days. Especially, the initial pH of Basic Kp was higher than that of BTSg, BTSa and Modena, and also higher than physiological pH of boar sperm (6.8∼7.5). When Basic Kp was added with various concentrations of EDTA (0, 0.63, 1.25 & 2.37g/L), Tris (0, 0.18, 0.35, 0.71 & 1.42g/L) and Citrate (0, 0.75, 0.81, 1.00, 1.25 & 1.50g/L) buffers for pH down-regulation and stabilization of pH, the group added with 1.25g EDTA, 1.42g Tris and 1.00g Citrate well maintained the neutral range of pH during storage (6.88 at day1 to 7.33 at day6 in Modified Kp), Especially, the concentrations of the buffers added in Modified Kp were lower, until 1/2∼1/4 ranges, than those in Modena and other extenders. The motility of frozen-thawed boar sperm diluted with Modified Kp was significantly higher than that of Basic Kp, BTSg, BTSa and Modena (87.0% vs. 55.0∼71.0% at day1; 13.3% vs. 0∼6.3% at day6), Conclusively, Modified Kp in this experiment was kept the favorable physiological conditions in spite of low concentrations of the buffers and motility of frozen-thawed boar sperm was obtained better than that of Basic Kp and other commercial products such as BTSg, BTSa and Modena.

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Inhibition of Enzymatic Degradation of Leucine Enkephalin and $[D-Ala^2]$-Leucine Enkephalinamide in Various Rabbit Mucosal Extracts by Inhibitors (효소 억제제에 의한 토끼의 점막 추출액중 로이신엔케팔린 및 [D-알라$^2$-로이신엔케팔린아미드의 분해 억제)

  • Chun, In-Koo;Park, In-Sook;Hyun, Jeen
    • Journal of Pharmaceutical Investigation
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    • v.26 no.3
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    • pp.175-185
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    • 1996
  • To inhibit the enzymatic degradation of leucine enkephalin (Leu-Enk) and its synthetic analog. $[D-ala^2]$-leucine enkephalinamide (YAGFL), in the nasal, rectal and vaginal mucosal and serosal extracts of rabbits, effects of enzyme inhibitors such as amastatin (AM), puromycin (PM), thiorphan (TP), thimerosal (TM), EDTA, N-carboxymethyl-Phe-Leu (CPL), phenylethyl alcohol (PEA), phenylmercuric acetate (PMA), benzalkonium chloride (BC) and modified cyclodextrins, alone or in combination, were observed by assaying the pentapeptides staying intact during incubation. Mucosa extracts were prepared by exposing freshly-excised mucosal specimens mounted on Valia-Chien cells to isotonic phosphate buffer while stirring. The degradation of Leu-Enk and YAGFL followed the apparent first-order kinetics. The half-lives (mean) in the nasal, rectal and vaginal mucosal extracts were found to be 1.07, 0.33 and 1.14 hr for Leu-Enk, and 16.9, 6.2 and 6.8 hr for YAGFL, respectively. AM or PM, which is an aminopeptidase inhibitor, did not show a sufficient inhibition of Leu-Enk $(50\;{\mu}g/ml)$ degradation in all kinds of extracts. $Dimethyl-{\beta}-cyclodextrin\;(DM-{\beta}-CyD)$ decreased the degradation rate constants of Leu-Enk about 2 or 3 times, comparing with no additive. However, the use of mixed inhibitors of AM $(50\;{\mu}M)$/TM (0.25 mM)/EDTA (5 mM) resulted in a full stabilization of Leu-Enk by decreasing the degradation rate constants 67.3, 161.3 and 113.8 times far the nasal, rectal and vaginal mucosal extracts, respectively, comparing with no inhibitor. With mixed inhibitors, Leu-Enk remained intact more than 90% after 6 hr-incubation. In the stabilization of YAGFL, hM, TP or CPL alone showed little efffct, and some additives demonstrated a considerable inhibition of YAGFL degradation in the rank order of TM > BC > EDTA. However, the addition of mixed inhibitors such as TM (0.5 mM) and EDTA (5 mM) into the extracts protected YAGFL from the degradation by more than 85% even after 24 hr-incubation, suggesting almost complete inhibition of YAGFL degradation in the extract. On the other hand, $DM-{\beta}-CyD\;or\;hydroxypropyl-{\beta}-cyclodextrin$ (10%) were also found to retard enzymatic degradation rates of YAGFL markedly, and resulted in staying intact more than 80% of YAGFL in the nasal and vaginal mucosal extracts, and more than 60% in the rectal mucosal extract after 16 hr-incubation.

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Stabilization of $Pb(Mg_{1/3}Ta_{2/3})O_3$ thin film by a thin $PbTiO_3$ seed layer and characterization of electric properties ($PbTiO_3$ 씨앗층을 이용한 $Pb(Mg_{1/3}Ta_{2/3})O_3$ 박막의 상안정화와 전기적 특성평가)

  • 김태언;유창준;문종하;김진혁
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2003.03a
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    • pp.211-211
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    • 2003
  • PbTiO$_3$ 씨앗층을 이용하여 완화형 강유전체 Pb(Mg$_{1}$3/Ta$_{2}$3/)O$_3$ (PMT) 박막의 페로브스카이트 상안정화와 열처리 조건에 따른 미세구조변화, 이에 따른 전기적 특성 변화에 관하여 조사하였다. PbTiO$_3$ 박막을 스핀코팅법으로 3000 rpm에서 20초간(111) 방향으로 배향된 Pt / Ti / SiO$_2$/ Si 기판에 증착하여 안정화된 페로브스카이트 박막을 얻었다. 이렇게 제조된 PbTiO$_3$를 Buffer 층으로 사용하고 그 위에 Pb(Mg$_{1}$3/Ta$_{2}$3/)O$_3$를 박막을 Spin coating방법으로 증착한 후, 급속열처리 방법(RTA)으로 550- $650^{\circ}C$ 사이에서 열처리하였다. 제조된 박막의 열처리 온도에 따른 미세구조 변화와 결정성을 XRD, SEM, TEM으로 분석하였고 박막의 저온 강유전 특성을 RT66A를 이용하여 평가하였다. Pb(Mg$_{1}$3/Ta$_{2}$3/)O$_3$ 박막의 경우 씨앗층이 없는 경우에는 pyrochlore상이 주상이었지만 씨앗층을 사용한 경우 페로브스카이트 상이 주상임을 확인하였고 열처리 온도가 증가할수록 페로브스카이트상의 상대적 양이 증가함을 확인하였다. 미세구조와 상의 변화에 따른 전기적 특성 변화에 관하여 자세하게 논의할 것이다.

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Determination of Horseradish Peroxidase (HRP) using an Enhanced Chemiluminescence Assay (증강 화학발광 기법을 이용한 horseradish peroxidase(HRP)의 검량)

  • Kim, Wongee;Kim, Keunhan;Lee, Seungmok
    • Journal of Korean Society on Water Environment
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    • v.25 no.1
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    • pp.84-89
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    • 2009
  • Our nation's water resources remain susceptible to contamination by phenolic agrichemicals. These compounds can be toxic to a variety of organisms including humans. Their disposal is restricted in many countries with strict limits for acceptable concentrations in drinking water. Enzyme-mediated in situ stabilization has been advocated as an approach for the treatment of phenolic compounds in soils and groundwater. This study reports the development of a new approach to quantify the activity of the HRP enzyme in aqueous systems. The method is based on the coupled processes of energy transfer and enhanced chemiluminescence using a luminol-$H_2O_2$-HRP system. In this study, the effects of solution pH, ionic strength and aqueous concentrations of HRP, $H_2O_2$ and enhancer were evaluated on the p-iodophenol-enhanced, HRP-catalyzed chemiluminescence reaction intensity in Tris-HCl buffer. All assay components were found to affect the maximum chemiluminescene intensity. The calibration curve for HRP showed the linear relationship with maximum light intensity.

Modification of Sweet Potato $\beta$-Amylase with Periodate-Oxidized Soluble Starch (과요소산 산화 전분에 의한 고구마 $\beta$-아밀라제의 수식)

  • 안룡근;지의상
    • The Korean Journal of Food And Nutrition
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    • v.3 no.2
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    • pp.123-132
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    • 1990
  • Sweet potato $\beta$-amylase is a tetrameric enzyme consisting of four identical polypeptide chains with a molecular weight of 5.6$\times$104, though most of the other $\beta$-amylases are monomeric enzymes. But, the relationship between subunit structure and catalytic function of the enzyme is not known. This study was done to know what the function of the subunit structure of the enzyme is. We obtained the monomer from the enzyme by the treatment of SDS, alkali pH buffer and urea. But the monomer had not activity. We tried to prepare the active monomer from the enzyme by the modification with periodate-oxidized soluble starch , In the result, we succeeded in isolating an active monomer as an oxidized soluble starch-conjugated form The active monomer had 57% of the original activity, 13.2% of the sugar and the molecular weight was estimated to be 5.4$\times$104. This results suggest that the tetrameric form of the enzyme is a most stable one and exists in nature, and the subunit structure of the enzyme Plays an important role in stabilization but not catalytic function.

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Effects of Salt and Heparin on the Activation of Lipoprotein Lipase(LPL)

  • Kim, Tae-Woong;Lee, Jae-Bok;Son, Heung-Soo;Kim, Sung-Wan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.355-361
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    • 1995
  • Lipoprotein lipase(LPL) is an acylglycerol hydrolase and is the extrahepatic enzyme responsible for the hydrolysis of triglyceride-rich plasma lipoproteins. LPL has been isolated from bovine milk by affinity chromatography on heparin-sepharose in 2M NaCl, 5mM barbital buffer, pH 7.4. Para-nitrophenyl butyrate(PNPB) was used as a substrate for the determination of LPL activity. Molecular weight of LPL was 55KD on 10% SDS-PAGE. When the effects of heparin on LPL activation were compared, LPL activity of heparin added group increased approximately 5 times higher than that of heparin non-added groups. These results indicated that heparin involved in the stabilization of LPL structure that led to increase enzyme activity. Furthermore, LPL activity increased about 4 times compared to the absence of heparin at various pH. LPL was stabilized when heparin was added either low or high salt concentrations. With the presence of heparin, NaCl concentration did not affect LPL activity at pH range 6∼9.

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Physicochemical Characteristics of Cephalosporin Derivative, CKD-604 : Stabilization and Solubilization in Aqueous Media (세팔로스포린계 유도체 CKD-604 물성연구 : 수용액중에서의 안정화 및 가용화)

  • Kwon, Soo-Yeon;Shin, Hee-Jong;Kim, Chong-Kook
    • Journal of Pharmaceutical Investigation
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    • v.29 no.3
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    • pp.205-210
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    • 1999
  • To formulate the parenteral delivery of a new cephalosporin derivative, 7-${\beta}$-[(2)-2-(2-arninothiazol-4-yl)-2methoxyiminoacetamido]- 3- [(2,3-cyclopenteno-4-carbamoyl-l-pyridinium)methyl]- 3-cephem-4-carboxylate sulfate( CKD604), the stability and solubility of CKD-604 in various aqueous media were investigated. The degradation kinetics of CKD-604 in aqueous solutions (ionic strength 0.1, pH 1-8) were studied at $37^{\circ}C$. The observed degradation rates followed pseudo first order kinetics. The pH-rate profile exhibited a minimum degradation rate at pH 5. The Arrhenius activation energy was 14.2 kcal/mol in pH 5 buffer solution. Excellent agreement between the cephalosporins' theoretical pH-rate profile and the experimental data indicated that the degradation pathway of CKD-604 could be predicted according to the general pathway of cephalosporins. The solubility of CKD-604 was 8.16 mg/ml at $25^{\circ}C$. To enhance the solubility and adjust the suitable pH, CKD-604 was solubilized by using sodium ascorbate, ascorbic acid and urea. The compositions were obtained to satisfy optimum pH and concentration, and the total amount of additives was several times of the active ingredient, CKD-604.

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Effect of stigmasterol in liposome bilayer on the stabilization of encapsulated ascorbic acid (리포솜 이중층의 stigmasterol이 포집된 ascorbic acid의 안정성에 미치는 영향)

  • Lee, Dong-Uk;Park, Hye-Won;Lee, Seung-Cheol
    • Korean Journal of Food Science and Technology
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    • v.52 no.2
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    • pp.200-203
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    • 2020
  • The effect of stigmasterol (SS), a phytosterol, in the liposome bilayer on the stability of encapsulated ascorbic acid (ASA) was evaluated. Liposomes, consisting of phosphatidylcholine (PC) and SS, and ASA were encapsulated by the dehydration/rehydration method. The average particle size of the liposome increased with increasing SS content. SS significantly increased the stability of encapsulated ASA. For example, ASA remaining in the liposomes of 100:0, 90:10, and 70:30 (PC:SS, w/w) ratios was 34.12%, 49.88%, and 58.58%, respectively, after storage for 8 days at 4℃, while only 7.66% ASA remained in the buffer under the same conditions. These results indicated that SS in liposomes increased the stability of encapsulated ASA.