• Title/Summary/Keyword: Cell Buffer

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Biodegradation of Bunker-C Oil by the Mixed Enrichment Culture of Marine Bacteria (혼합배양 해양세균에 의한 Bunker-C유의 생물분해)

  • PARK In-Sick;PARK Jung-Youn;SUH Kuen-Hack;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.2
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    • pp.152-156
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    • 1987
  • A mixed population of marine bacteria was obtained to catalize the biodegradation of bunker-C fuel oil by means of the enrichment culture technique. Samples used for the enrichment culture were collected from sea water and sediments in the vicinity of Pusan, Chungmu, and Ulsan in Korea. As the biodegradation of bunker-C oil proceeded, the number of bacteria increased from $1.1\times10^6\;to\;8.7\times10^8$ cells per ml when pH was bufferized by 0.1 M Tris-HCl buffer to 7.6, then oil dispersion increased to $OD^{540}$ 2.2 and approximately $48\%$ of the oil was biodegradated in 10 days. Oil dispersion was absolutely dependent on the addition of nitrogen and phosphate sources in sea water. High and low sulfur-containing bunker-C and crude oil could be dispersed similarly. Bunker-C oil was dispersed rapidly at the pH ranging from 7.0 to 8.0 and dispersed to the amount of 7.5 g per liter of sea water medium.

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Expression and Characterization of Calcium- and Zinc-Tolerant Xylose Isomerase from Anoxybacillus kamchatkensis G10

  • Park, Yeong-Jun;Jung, Byung Kwon;Hong, Sung-Jun;Park, Gun-Seok;Ibal, Jerald Conrad;Pham, Huy Quang;Shin, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.606-612
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    • 2018
  • The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-${\beta}$-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was $80^{\circ}C$ and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at $60^{\circ}C$. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of $Ca^{2+}$ and $Zn^{2+}$ ions. The kinetic properties, $K_m$ and $V_{max}$, were calculated as 81.44 mM and $2.237{\mu}mol/min/mg$, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.

Microfluidic System for the Measurement of Cupric Ion Concentration using Bilayer Lipid Membrane on Silver Surface (은 표면의 이중층 지질막에 의한 구리 이온 농도 측정용 마이크로플루이딕 시스템)

  • Jeong, Beum Seung;Kim, Do Hyun
    • Korean Chemical Engineering Research
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    • v.48 no.1
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    • pp.33-38
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    • 2010
  • A microfluidic system has been developed using biomaterial for the measurement of cupric ion concentration. The cell-membrane-mimicking bilayer lipid membrane(BLM)-coated silver electrode was used for the sensing of cupric ion concentration. The silver-supported BLM could increase its stability. A silver-supported bilayer lipid membrane(s-BLM) was easily obtained using its self-assembling characteristics by immersing silver wire into lipid(phosphatidylcholine; PC) solution and then dipping into aqueous KCl solution. These s-BLMs were used to determine the relationship between $Cu^{2+}$ concentration and current crossing s-BLM. Their relationship showed high linearity and reproducibility. The calibration curve was constructed to express the relationship between $Cu^{2+}$ concentration and current in the $Cu^{2+}$ concentration range of 10 and $130{\mu}M$. This calibration curve was used to measure $Cu^{2+}$ concentration in an unknown sample. Microfluidic system with s-BLM was made of PDMS(polydimethyl siloxane) using typical soft photolithography and molding technique. This integrated system has various functions such as activation of the silver surface without cutting silver wire, coating of BLM on silver surface, injection of KCl buffer solution, injection of $Cu^{2+}$ sample and measurement of $Cu^{2+}$ concentration in the sample.

The Microstructure of Magnetite Coated on Honeycomb and Characteristics of CO2 Decomposition (허니컴에 코팅한 마그네타이트의 미세구조 및 CO2 분해특성)

  • 윤용운;김은배;이병하;고태경;오재희
    • Journal of the Korean Ceramic Society
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    • v.41 no.5
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    • pp.410-416
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    • 2004
  • In this study, we fabricated magnetite coated on a cordierite honeycomb which has complex shape by ultrasound-enhanced ferrite plating. The effects of the plating condition on the formation of the magnetite and its microstructure were investigated. The magnetite coated on the honeycomb became an oxygen-deficient ferrite by H$_2$ gas reduction, then the effects of the molar concentrations of ammonium acetate for $CO_2$ gas decomposition have been studied. As the molar concentration of a pH buffer($CH_3$COONH$_4$, 0.1946∼0.3892 M) solution increased, the average particle size increased about 200∼250 nm. The magnetite coated on the honeycomb was reduced by H$_2$ gas for 2 h at 30$0^{\circ}C$. The inner pressure change in the cell began to occur at 315∼34$0^{\circ}C$. The H$_2$-Reduced magnetite coated on the honeycomb at 35$0^{\circ}C$ contained an oxygen deficient magnetite and $\alpha$-Fe phase. The thermogravimetric analysis with H$_2$ reduction and $CO_2$ decomposition were carried out with the magnetite coated on the honeycomb. A weight loss in process of H$_2$ reduction occurred between 32$0^{\circ}C$ and 34$0^{\circ}C$, while a weight gain was observed during the $CO_2$ decomposition.

Use of Bacteriocin Produced by Lactococcus sp. CU216 with pH Sensitive Liposome Entrapment (Lactococcus sp. CU216이 생산하는 박테리오신을 함유한 pH Sensitive Liposome의 응용)

  • 박성수;김명희;한경식;오세종
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.97-102
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    • 2004
  • The objective of this study was to control Kimchi fermentation using pH sensitive bacteriocin entrapping liposome(bacteriocin-liposome). The liposomes were prepared by the reverse-phase evaporation method from a mixture of DPPC(dipalmitoyl phosphatidylcholine, DPPE(dipalmitoyl phosphatidylethanolamine), DOPC(dioleoyl phosphatidylcholine) and cholesterol in a molar ration of 4:2:1:4. The bacteriocin-liposome was disruptured at pH 4 of buffer and was stable at alkaline pHs(6 and 7). Irrespective of the addition of the bacteriocin-liposomes, the pH of every Kimchi sample decreased to 5 during 5 days storage at 5$^{\circ}C$. Kimchi samples treated with bacteriocin-liposomes maintained pH 4 or higher, while Kimchi samples not treated with bacteriocin-liposomes exhibited pH 3.58 or lower. In general, the pH of Kimchi samples stored at 20$^{\circ}C$ decreased faster, compared to that of Kimchi samples stored at 5$^{\circ}C$. The pH of Kimchi samples treated with the bacteriocin-liposomes was 3.9 during 90 days storage, while that of the samples not treated with the bacteriocin-liposomes was 3.68 and 3.32 during 30 days and 90 days storages, respectively. Lactic acid bacteria in Kimchi treated with the bacteriocin-liposome grew relatively slow at 5$^{\circ}C$. The viable cell number of lactic acid bacteria increased up to 4${\times}$10$\^$7/ cells/ml and then decreased to 8${\times}$10$\^$6/ cells/ml during 90 days storage at 5$^{\circ}C$.

미토콘드리아 미세구조의 변형과 생쥐 초기 배아의 발생능력과의 연관성에 대한 연구

  • 전진현;임천규;궁미경;고경남;김문규
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.106-106
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    • 2002
  • 미토콘드리아는 세포내의 에너지대사에서 중요한 역할을 수행하는 세포내 소기관이며, 자체의 유전물질이 모계를 통해 유전되는 특징을 가지고 있다. 포유류 초기 배아의 발생과정에서 미토콘드리아의 역할과 기능에 대한 연구는 미진한 상태이다. 본 연구에서는 생쥐 초기 배아의 발생과정에서 관찰할 수 있는 미토콘드리아 미세구조의 변화 양상을 살펴보고, 이와 초기 배아의 발생 능력과의 관련성을 밝혀보고자 하였다. 과배란 유도된 ICR 생쥐로부터 배란된 난자와 2-세포기 배아를 수획하여 76 배양액으로 포배기까지 체외배양하면서, 각각의 발생단계에 따라 시료를 수획하였다. 미토콘드리아 미세구조의 변화는 일반적인 투과전자현미경방법(TEM)을 이용하여 관찰하였다. 미토콘드리아의 미세구조는 배란 난자에서 4-세포기 배아까지는 구형이고 크리스타가 발달하지 않은 원시형태였지만, 포배기로 발달함에 따라 크리스타가 발달된 막대형의 전형적인 미토콘드리아로 분화됨이 관찰되었다. 체외배양 중에 발생이 지연되거나 정지된 배아에서 관찰한 미토콘드리아의 미세구조는 공포화 (vacuolization), 크리스타 발달 지연, 손상된 미토콘드리아의 세포막 등과 같은 비정상적인 변형을 관찰할 수 있었다. 또한, 극체에 존재하는 미토콘드리아의 미세구조는 정상적인 핵내의 유전자와의 상호작용이 없어 미분화 상태로 포배기까지 유지되는 것을 관찰하였다. 이상의 결과를 통해 미토콘드리아의 정상적인 분화 과정이 초기 배아의 발생능력과 관련되어 있음을 알 수 있었으며, 포유류 초기 배아의 체외배양시스템을 개선하는데 미토콘드리아 미세구조의 관찰과 변화에 대한 고려가 있어야 될 것으로 생각된다. buffer A 용액으로 세척하여 유리 petri dish의 바닥에 부착된 macrophage만을 cell scraper로 분리하였다. 분리한 macrophage는 0.5-1 $\times$ $10^{6}$ cells/$m\ell$가 되게 조정하여, IL-I 을 0.001, 0.01, 0.1 또한 1 ng/$m\ell$를 첨가하여 농도에 따른 효과를 조사하였고, 각각 24, 48, 72, 96 또한 120시간을 배양하여 시간에 의한 효과도 실시하였다. 각 배치구에서 얻어진 배양액은 TGF-$\beta$를 조사하기 전까지 -2$0^{\circ}C$에 동결 보존하였다. TGF-$\beta$의 측정은 TGF-$\beta$ kit(promega, USA)를 이용하여 실시하였으며, 통계학적 분석은 Anova test를 Statview program을 이용하여 분석하였다. 시험의 결과 대조구에 비해 IL-I 첨가구는 2-3배의 TGF-$\beta$생산을 보였으며, 배양시간에 따른 생산은 시간이 지남에 따라 약간 상승하는 경향을 보였으나, 유의적인 차이를 보이지는 않았다. 또한 IL-I의 농도에 따른 생산의 변화는 IL-I의 농도에 따라 약간의 차이를 보였고 유의적인 차이는 인정되지 않았다. 임신 및 비임신의 경우 임신우의 비장 macrophage가 비임신보다는 약간 상승하는 거스로 나타났다. 이상의 결과로 볼 때 IL-I $\alpha$$\bet

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소 비장유래 macrophage의 체외배양시 IL-I$\alpha$가 TGF-$\beta$의 생산에 미치는 영향

  • 최선호;성환후;장유민;이장희;연성흠;류일선;손동수;유충현
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.88-88
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    • 2002
  • 인공수정 및 수정란기술의 활성화에 따라 소에 있어서 인공수정은 90%이상이 실시되고 있으나, 수정란이식은 수정란의 생산이 안정적이지 않아 활성화에 많은 지장을 초래하고 있다. 이의 원인은 수정란이식에 의한 수태율의 저하가 가장 크며, 수태율 향상을 위하여 수란우에 progesterone, hCG 등의 주사가 실시되고 있다. 그러나 이는 수정란의 착상에 있어서 자궁의 환경을 개선한다고 하나, 착상의 정확한 기전의 구명은 미미한 상태이다. 한편 비장유래 macrophage가 황체를 자극하고 TGF-$\beta$의 생산을 유도하는 것으로 보고되고 있으며, IL-I $\alpha$$\beta$에 따라 TGF-$\beta$ 생산에 있어서 약간의 차이를 보이는 것으로 보고되고 있다. 따라서 본 연구는 비장유래 macrophage가 TGF-$\beta$의 생산시 임신관련 Cytokine인 IL-I$\alpha$와의 관계를 조사하기 위하여 실시하였다. 임신 및 비임신 도축 암소의 비장을 채취하여 얼음에 채워 실험실로 운반한 후 비장의 표면을 70%의 알콜로 세척하고, 표피를 벗겨 비장조직을 세절하여 10% FBS+DMEM에 넣어 조직을 눌러 짜면서 조직속의 세포를 분리하였다. 세척한 배양액은 4-5$m\ell$를 100mm 유리 petri dish에 넣고 39$^{\circ}C$, 5% $CO_2$, 95% 공기인 배양기에서 2시간이상 배양하였으며, 배양 후 냉장된 buffer A 용액으로 세척하여 유리 petri dish의 바닥에 부착된 macrophage만을 cell scraper로 분리하였다. 분리한 macrophage는 0.5-1 $\times$ $10^{6}$ cells/$m\ell$가 되게 조정하여, IL-I 을 0.001, 0.01, 0.1 또한 1 ng/$m\ell$를 첨가하여 농도에 따른 효과를 조사하였고, 각각 24, 48, 72, 96 또한 120시간을 배양하여 시간에 의한 효과도 실시하였다. 각 배치구에서 얻어진 배양액은 TGF-$\beta$를 조사하기 전까지 -2$0^{\circ}C$에 동결 보존하였다. TGF-$\beta$의 측정은 TGF-$\beta$ kit(promega, USA)를 이용하여 실시하였으며, 통계학적 분석은 Anova test를 Statview program을 이용하여 분석하였다. 시험의 결과 대조구에 비해 IL-I 첨가구는 2-3배의 TGF-$\beta$생산을 보였으며, 배양시간에 따른 생산은 시간이 지남에 따라 약간 상승하는 경향을 보였으나, 유의적인 차이를 보이지는 않았다. 또한 IL-I의 농도에 따른 생산의 변화는 IL-I의 농도에 따라 약간의 차이를 보였고 유의적인 차이는 인정되지 않았다. 임신 및 비임신의 경우 임신우의 비장 macrophage가 비임신보다는 약간 상승하는 거스로 나타났다. 이상의 결과로 볼 때 IL-I $\alpha$$\beta$subunit 보다 TGF-$\beta$ 생산에 있어서 서로 다른 양상을 보일 것으로 추정되며, IL-I은 macrophage의 직접적인 영향을 주기보다는 황체세포를 매개로 한 자궁에 TGF-$\beta$의 생산을 유도하는 것으로 사료되며, 임신관련 cytokine에 대한 다양한 연구가 요구되고 있다.

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Sulfate Modulation for Hydrogen Production by Chlamydomonas reinhardtii in Continuous Culture (Chlamydomonas reinhardtii 연속 배양에서 수소생산을 위안 황 조절)

  • Kim, Jun-Pyo;Park, Tai-Hyun;Kim, Mi-Sun;Sim, Sang-Jun
    • KSBB Journal
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    • v.20 no.6
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    • pp.453-457
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    • 2005
  • We investigated the effect of sulfate re-addition on hydrogen production under sulfur-deprived condition. When the final concentration of sulfate to cell suspensions($0{\sim}120{\mu}M$) was increased, chlorophyll concentration, culture density, and total amount of $H_2$ produced, increased up to an optimal concentration of $30{\mu}M\;MgSO_4$. Maximum hydrogen volume was 236 mL $H_2/L$ culture at $30{\mu}M\;MgSO_4$. However, the addition of excess sulfate(above $MgSO_4\;60{\mu}M$) delayed the start of hydrogen production and the induction of hydrogenase. Accordingly, the final yield of hydrogen production was reduced. Using these results, we attempted the continuous and sustained hydrogen production by sulfate re-addition($30{\mu}M\;MgSO_4$) using a single C. reinhardtii culture for up to 4 cycles. In total, hydrogen production volume was 625 mL $H_2/L$ culture.

Graphene Quantum Dot Interfacial Layer for Organic/Inorganic Hybrid Photovoltaics Prepared by a Facile Solution Process (용액 공정을 통한 그래핀 양자점 삽입형 유/무기 하이브리드 태양전지 제작)

  • Kim, Youngjun;Park, Byoungnam
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.6
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    • pp.646-651
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    • 2018
  • This paper reports that the electronic properties at a $P3HT/TiO_2$ interface associated with exciton dissociation and transport can be tailored by the insertion of a graphene quantum dot (GQD) layer. For donor/acceptor interface modification in an $ITO/TiO_2/P3HT/Al$ photovoltaic (PV) device, a continuous GQD film was prepared by a sonication treatment in solution that simplifies the conventional processes, including laser fragmentation and hydrothermal treatment, which limits a variety of component layers and involves low cost processing. The high conductivity and favorable energy alignment for exciton dissociation of the GQD layer increased the fill factor and short circuit current. The origin of the improved parameters is discussed in terms of the broad light absorption and enhanced interfacial carrier transport.

Inflammation Scan Using $^{99m}Tc-HMPAO$ Labelled Leukocytes ($^{99m}Tc-HMPAO$를 이용한 자가백혈구표지 및 그를 이용한 염증병소의 스캔)

  • Yang, Woo-Jin;Chung, Soo-Kyo;Shinn, Kyung-Sub;Bahk, Yong-Whee;Kim, Hoon-Kyo
    • The Korean Journal of Nuclear Medicine
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    • v.23 no.2
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    • pp.219-223
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    • 1989
  • Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.

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