• Title/Summary/Keyword: buffer stabilization

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Studies on the Simplified Hemagglutination Reaction to Pasteurella multocida and the Stabilization of Erythrocytes (Pasteurella multocida에 대(對)한 간이적혈구(簡易赤血球) 응집반응(凝集反應)과 적혈구(赤血球)의 안정화(安定化)에 관(關)한 연구(硏究))

  • Lee, Hak Cheul;Chung, Yung Gun;Kim, Kyo-Joon
    • Korean Journal of Veterinary Research
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    • v.10 no.1
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    • pp.11-23
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    • 1970
  • Recently Carter(1952) reported the capsule antigens of Pasteurella multocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test. A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy. The author tried adapting Carter's hemagglutination method to the slide method so called "micromethod technique", and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes. The results obtained are summarized as follows: 1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter's hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner's microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding $0.05m{\ell}$ of antigen-sensitized erythrocytes suspension diluted to one percent on $0.05m{\ell}$ of serially diluted antibody-containing sera, and the final result can be read after 60 minutes at the room temperature ($15^{\circ}C$). 2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination. 3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction. 4. The antigen-sensitization against erythrocytes at $37^{\circ}C$ is optimal for the slide hemagglutination. 5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes. 6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes. 7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination. 8. The erythrocytes preserved at $4^{\circ}C$ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving. 9. The stable preparation of erythrocytes can be obtained by treating the cells at $37^{\circ}C$ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at $4^{\circ}C$ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at $4^{\circ}C$. 10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity. 11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes. 12. There is no difference between the saline and the isotonic buffer solution on the reaction of hemagglutination.

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An Investigation of Treatment Effects of Limestone and Steel Refining Slag for Stabilization of Arsenic and Heavy Metal in the Farmland Soils nearby Abandoned Metal Mine (폐금속 광산 주변 비소 및 중금속 오염농경지의 안정화 처리를 위한 석회석과 제강슬래그의 처리효과 검토)

  • Yun, Sung-Wook;Kang, Sin-Il;Jin, Hae-Geun;Kim, Ha-Jin;Lim, Young-Cheol;Yi, Ji-Min;Yu, Chan
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.734-744
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    • 2011
  • A soil stabilization method is an effective and practical remediation alternative for arsenic (As) and heavy metal contaminated farmland soils nearby abandoned metal mine in Korea. This method is a technique whereby amendments are incorporated and mixed with a contaminated soil. Toxic metal bind to the amendments, which reduce their mobility in soil, so the successful stabilization of multi-element contaminated soil depends on the combination of critical elements in the soil and the type of amendments. The objective of this study is to investigate the treatment effects and applicability of limestone (LS) and steel refining slag (SRS) as the amendment for farmland soil contaminated with As and heavy metals, and a lab-column test was conducted for achieving this purpose. The result showed that soil treated with LS and SRS maintained pH buffer capacity and, as a result, the heavy metal leaching concentration was quite low below the water quality standard compared to untreated soil which leachate exceeding the water quality standard was observed, however, the arsenic concentration rather increased with increasing mixture ratio of SRS. This was believed to be related to phosphorus (P) contained in SRS, and dominancy in the competitive adsorption relation between As and P binding strongly to iron might be different according to soil characteristic. We suggested that LS is a effective amendment for reducing heavy metals in soil, and SRS should be used after investigating its applicability based on the adsorption selectivity of arsenic and phosphorus in selected soil.

Spheroplast Formation, Regeneration and Fusion of Flavimonas oryzihabitans KU21 (Flavimonas oryzihabitans KU21의 원형질체 생성, 재생 및 융합)

  • 이수연;임영복;박용근;이영록
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.318-325
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    • 1993
  • The optima] conditions for the formation, the regeneration. and the spheroplast fusion of Flavimonas aryz/habitans spheroplasts were investigated. Cells were transformed to spherop]asts effectively by treatment of 0.5% volume (v/v) of 0.] M EDTA and ]00 flg/ml lysozyme at $37^{\circ}C$ for 30 min without shaking. Magnesium chloride and calcium chloride were effective on the stabilization of spheroplasts. and 20 mM calcium chloride in the rich regeneration medium improve the yield of regenerants as much as 3.5-fo]d. Addition of 0.8% bovine serium albumine (BSA) in dilution buffer for spheroplast formation improved the stabilization of spheroplasts over extended periods (4-6 hr) at room temperature. and thus increased the yield of recombinants to 4.5-fold. The spheroplast formation frequency and regeneration frequency of F aryzihabitans strain was 90.10% and 3.800/." respectively. The first regenerated cell of F. aryzihabitans spheroplasts were appeared 6 hours after plating. By I I hours after plating, 80% of spheroplasts were regenerated on thc rich regeneration medium containing 0.5 M sucrose. The intraspeci11c spheroplast fusion of F urvz/habitans was carried out and the properties of obtained fusants were investigated. Formation of fusion products was effective when the Flav/munas spheroplast mixture was treated with 40%(w/v) PEG6000 and 20 mM CaCl, for 10 min at room temperature. and thc formation of frequency of recombinants were $2.0{\times}10^{-5}~3.6{\times}10^{-5}$. All tested recombinant clones were very stable on further propagation.

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A Study on the Stabilization of the Papain Enzyme in the Moderately Concentrated Anionic Surfactant System (음이온 계면활성제에서 파파인 효소의 안정도에 관한 연구)

  • Kim, Ji-Yeong;Kim, Jin-Woo;Kim, Yong-Jin;Lee, Jae-Wook;Lee, Hae-Kwang;Kang, Hak-Hee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.2
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    • pp.93-97
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    • 2007
  • Even in the moderately concentrated anionic surfactant system, some special encapsulation method can shield the papain enzyme from proteolytic attacks. The stabilization of enzyme has been a major issue for successful therapies. In this study, we first stabilized an enzyme, papain in the microcapsules by using polyols, polyethyleneglycol (PEG), poly-propyleneglycol (PPG), and PEG-PPG-PEG block copolymer. In the analysis of EDS and CLSM, it was demonstrated that polyols are effectively located in the interface of papain and polymer. Polyols located in the interface had an ability to buffer the external triggers by hydrophobic partitioning, preventing consequently the catalytic activity of papain in the micro-capsules. Second. we introduced multi-layer capsulation methods containing ion complex. Such a moderately concentrated anionic surfactant system as wash-off cleansers, surfactants and waters can cause instability of entrapped enzymes. Surfactants and water in our final products swell the surface of enzyme capsules and penetrate into the core so easily that we can not achieve the effect of enzyme, papain. In this case, the ion complex multi-layer capsule composed of sodium lauroyl sarcosinate and polyquaternium-6 could effectively prevent water from penetration into the core enzyme, followed by in vivo test, and evaluate the stratum corneum (SC) turn-over speed.

Preparation and Characterization of Ethosome Containing Hydrophobic Flavonoid Luteolin (소수성 플라보노이드인 루테올린을 함유한 Ethosome의 제조 및 특성조사)

  • Lee, Sang Min;Choi, Moon Jae;Lee, Young Moo;Jin, Byung Suk
    • Applied Chemistry for Engineering
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    • v.21 no.1
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    • pp.40-45
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    • 2010
  • Entrapment of hydrophobic flavonoid luteolin into ethosome was carried out for improving its stability and making practical application in the field of drug and cosmetics. The formation of liquid crystalline phase and its thermal properties were investigated by polarized optical microscopy and DSC. The phase inversion from W/O to W/O/W was detected by conductivity change with the addition of PBS buffer solution into the ethanol-dissolved lecithin mixture. The particle size change of ethosome with constituent composition was examined, which showed that the incorporation of luteolin into lecithin up to 10% had little effect on the size of ethosome. Enhancement of stability of luteolin by entrapment into ethosome was verified through DPPH test. The stabilization efficacy of ethosome was improved further by the addition of tocopherol.

Evaluation of the Impacts of Water Quality Management in Kyongan Stream Watershed using SWAT Model (SWAT 모델을 이용한 경안천 유역의 수질관리 영향 평가)

  • Jang, Jae-Ho;Yoon, Chun-Gyeong;Jung, Kwang-Wook;Kim, Hyung-Chul
    • Journal of Korean Society on Water Environment
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    • v.26 no.3
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    • pp.387-398
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    • 2010
  • SWAT model would be applied to evaluate the pollutant removal capacity with various best management practices (BMPs) in Kyongan stream watershed which plays an important role in water quality conservation and improvement of Paldang reservoir. The methods for the representation of various BMPs scenarios with SWAT is developed and evaluated. Riparian buffer strip, agricultural conservation practices to reduce fertilizer, sediment, and nutrients occurring from farm field (Grassed swale, Contour farming/Parallel terrace, Field border, Farm retention pond, Grade stabilization structure), and washland such as wetland and pond to extend detention and improve water quality are represented in SWAT. And to represent the expansion of existing Waste Water Treatment Plants (WWTPs) in Soil and Water Assessment Tool (SWAT), reduction effect for point source pollutants was simulated. As the result of simulation, the removal rates of SS, TN, TP from scenarios of Kyongan stream watershed are the average annual SS yield by 5.2% to 69.2%, the average annual TN yield by 0.5% to 26.3%, and the average annual TP yield by 1.3% to 32.5%, respectively. This study has demonstrated that the SWAT is a very reliable and useful water quality and quantity assessment tool, and the BMPs representation in SWAT for watershed management is able to effectively simulate in Kyongan Stream watershed.

Conformational Study of Human Serum Albumin in Pre-denaturation Temperatures by Differential Scanning Calorimetry, Circular Dichroism and UV Spectroscopy

  • Rezaei-Tavirani, Mostafa;Moghaddamnia, Seyed Hassan;Ranjbar, Bijan;Amani, Mojtaba;Marashi, Sayed-Amir
    • BMB Reports
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    • v.39 no.5
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    • pp.530-536
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    • 2006
  • Thermal conformational changes of human serum albumin (HSA) in phosphate buffer, 10 mM at pH = 7 are investigated using differential scanning calorimetric (DSC), circular dichroism (CD) and UV spectroscopic methods. The results indicate that temperature increment from $25^{\circ}C$ to $55^{\circ}C$ induces reversible conformational changes in the structure of HSA. Conformational change of HSA are shown to be a three-step process. Interestingly, melting temperature of the last domain is equal to the maximum value of fever in pathological conditions, i.e. $42^{\circ}C$. These conformational alterations are accompanied by a mild alteration of secondary structures. Study of HSA-SDS (sodium dodecyl sulphate) interaction at $45^{\circ}C$ and $35^{\circ}C$ reveals that SDS affects the HSA structure at least in three steps: the first two steps result in more stabilization and compactness of HSA structure, while the last one induces the unfolding of HSA. Since HSA has a more affinity for SDS at $45^{\circ}C$ compared to $35^{\circ}C$, It is suggested that the net negative charge of HSA is decreased in fever, which results in the decrease of HSA-associated cations and plasma osmolarity, and consequently, heat removal via the increase in urine volume.

A P2P Real-time Game System for Multiplayer on Overlay Network (Overlay Network망에서의 실시간 멀티플레이어 P2P게임 시스템)

  • Jung, Mi-Sook;Park, Kyoo-Seok
    • Journal of Korea Multimedia Society
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    • v.13 no.1
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    • pp.38-46
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    • 2010
  • A stable game managing system is absolutely needed to accept simultaneous interlacing of many users for on- line game system. The proposed P2P on-line game system in this paper is able to get stable and real-time game managing within limited time stamp utilizing through reorganizing peers according to synchronous time which can be avoid congestion on one region, it is possible to synchronize for game nodes within limited time stamp utilizing. Reorganizing M-tree which leads 10 distribute loads. The system manages each region unit, and it is execute no matter how big game sizes are. Thus to ensure such as the problems of expand server and stabilization or message transmission. Also, prove efficiency of the suggested system through the simulation.

Detection of C-Reactive Protein Using Direct-binding Quartz Crystal Microbalance Immunosensor (직접결합방식 수정진동자 면역센서에 의한 C-Reactive Protein 검출)

  • Kim, N.;Kim, D.K.;Cho, V.J.
    • KSBB Journal
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    • v.22 no.6
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    • pp.443-446
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    • 2007
  • A prognostic indicator of coronary heart disease, C-reactive protein, was tried to be determined by a batch-type quartz crystal microbalance immunosensor. The sensor was operated by direct-binding mode and the optimum concentration for the corresponding antibody for immobilization was $50{\mu}g/ml$. The reaction buffer for the system was 0.1 M sodium phosphate (pH 7.0) and system operation was performed in the order of baseline stabilization, analyte addition and measurement, and regeneration of the sensor chip with 10 mM NaOH. When plotted in double-logarithmic scale, the sensor showed a linear detection range of 0.27-106.00 nM for rat C-reactive protein with the limit of detection of 0.53 nM. It also showed a good reusability.

The Spheroplast Formation and Regeneration of Pseudomonas spp. (Pseudomonas spp.의 원형질체 생성과 재생)

  • 이주실;이영록
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.24-31
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    • 1986
  • The optimal conditions for the formation and the regeneration of Pseudomonas spheroplast were measured. Pseudomonas spp. cells were transformed to spheroplasts from 99.0% to 99.9% by treatment of $100{\mu}g/ml$ lysozyme and 10mM EDTA at room temperature. The optimal pH for the spheroplast formation was pH 8.0. Magnecium chloride, calcium chloride and streptomycin were effective on the stabilization of Pseudomonas spheroplast, while $Mg^+\;and\;Na^+$ ions were effective on the formation of Pseudonomas spheroplast. Rich Regeneration Medium was used for the regeneration of Pseudonomas spheroplast. To improve regeneration frequency, Bovin Serum Albumine and cationic ions were added to the spheroplast dilution beffer and regeneration environment respectively. Treatment of 20mM calcium chloride in ehr Rich Regeneration Medium could improve the yield of regenerants as much as 28-fold. Treatment of 1% Bovin Serum Albumine in the spgeroplast formation and dilution buffer increased the yield of regenerants to 10-fold. Also, the regeneration frequency was improved to 14-fold shen Rich Regeneration Meidum containing 0.5% Gelatin was used for regeneration as well as 1% Bovin Serum Albumine.

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