• Title/Summary/Keyword: Ni-Cd

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Soluble Expression of a Human MnSOD and Hirudin Fusion Protein in Escherichia coli, and Its Effects on Metastasis and Invasion of 95-D Cells

  • Yi, Shanze;Niu, Dewei;Bai, Fang;Li, Shuaiguang;Huang, Luyuan;He, Wenyan;Prasad, Anand;Czachor, Alexander;Tan, Lee Charles;Kolliputi, Narasaiah;Wang, Feng
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1881-1890
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    • 2016
  • Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

Low Temperature Sintering of PNN-PZT Ceramic for Piezoelectric Generator and Its Piezoelectric Properties (압전 발전시스템 개발을 위한 PNN-PZT 세라믹스의 저온소결 및 압전특성 평가)

  • Lee, Myung-Woo;Kim, Sung-Jin;Yoon, Man-Soon;Ryu, Sung-Lim;Kweon, Soon-Yong
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.06a
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    • pp.306-306
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    • 2008
  • 기계적 에너지를 전기적 에너지로 변환하는 에너지 변환소자인 압전 세라믹스는 액추에이터, 변압기, 초음파모터, 초음파 소자 및 각종 센서로 응용되고 있으며, 그 응용분야는 크게 증가하고 있다. 최근에는 이러한 압전 소자를 앞으로 도래하는 ubiquitous, 무선 모바일 시대의 휴대용 전자제품, robotics, MEMS 분야 등의 대체 에너지원으로 응용하기 위한 연구가 진행되고 있다. 특히 인간의 걷기 운동 등과 같은 일상적인 동작으로 필요한 전력을 얻을 수 있고, 세라믹 소자를 이용하기 때문에 전자노이즈가 발생되지 않을 뿐 아니라 반영구적으로 사용할 수 가 있어서, 기존 이차전지를 대체 또는 보완 할 수 있는 방안도 검토되고 있다. PZT계 세라믹스는 높은 유전상수와 우수한 압전특성으로 전자세라믹스 분야에서 가장 널리 사용되어지고 있지만 $1200^{\circ}C$ 이상의 높은 소결온도 때문에 $1000^{\circ}C$ 부근에서 급격히 휘발되는 PbO로 인한 환경오염과 기본조성의 변화로 인한 압전 특성의 저하가 문제시 되고 있다. 또한 적층 세라믹스의 제작 시 구조적 특성상 내부전극이 도포된 상태에서 동시 소결이 필요한데, 융점이 낮은 Ag전극 대신 값비싼 Pd나 Pt가 다량 함유된 Ag/Pd, Ag/Pt 전극이 사용되고 있어 경제성이 떨어지는 단점을 갖게 된다. 순수 Ag 전극을 사용하거나 Ag의 비율이 높은 내부전극을 사용하기 위해서는 $900^{\circ}C$ 이하에서 소결되고 우수한 전기적 특성을 보이는 압전 세라믹스를 개발 하는 것이 필요하다. 따라서 본 연구에서는 압전특성이 우수한 $(Pb_{1-x}Cd_x)(Ni_{1/3}/Nb_{2/3})_{0.25}(Zr_{0.35}/Ti_{0.4})O_3$ 계의 조성을 설계하고, 소걸온도를 낮추기 위해서 2 단계 하소법을 이용하였다. 또한 $MnCO_3$, $SiO_2$, $Pb_3O_4$ 등을 소량 첨가하여 액상 소결 특성을 부여하여 소결 온도를 감소시키려는 시도도 하였다. 분말을 볼 밀링 (ball milling)을 통해 24시간 동안 혼합하고, 혼합된 분말은 $800^{\circ}C$에서 2시간 동안 하소하였다. 하소한 분말을 다시 72시간 동안 볼 밀링 하여 최종 분말을 얻었다. 최종 분말에 PVB를 첨가하여 직경 15mm의 디스크 형태로 성형한 후, 850~$975^{\circ}C$ 범위에서 온도를 변화시키면서 소결을 하였다. 최종 분말 및 소결된 시편을 XRD분석을 통하여 상을 확인하였고, SEM을 이용하여 미세조직을 관찰 하였다. 전기적 특성을 평가하기 위하여 두께를 1mm로 연마한 시편에 Ag 전극을 도포하여 $650^{\circ}C$에서 열처리한 후, 분극처리 하였다. 압전특성은 $d_{33}$-meter로 측정하였고, impedance analyzer를 이용하여 압전 특성 (전기기계결합계수 및 기계적품질계수)을 측정 하였다. 또한 강유전체 특성 평가 장치 (Precision-LC)를 이용하여 분극-전계 특성을 평가하였다. 이상의 연구를 통하여 소결 온도가 $900^{\circ}C$인 경우에서도 양호한 압전 특성을 확보 할 수 있었다.

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Population and transfer factor of basidiomycota collected in the heavy metal-contaminated and healthy soils (중금속 오염지 및 건전지역에서 수집된 담자균류의 분포)

  • Jang, Kab-Yeul;Choi, Sun-Gyu;Lee, Kang-Hyo;Seok, Soon-Ja;Cheong, Jong-Chun;Jung, Gu-Bok;Kim, Gyu-Hyun;Sung, Jae-Mo
    • Journal of Mushroom
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    • v.3 no.4
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    • pp.145-153
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    • 2005
  • Two hundred seventy-four microorganisms were isolated from the soil of abandoned mines around for selecting the heavy metal-degrading strains and their microbial diversity was analyzed. The specimens of macrofungi were isolated from the soil of abandoned mines around for selecting the heavy metal-degrading strains. Eunseong and Dogok in Gyeongsangbuk-do, for selecting the heavy metal-degrading strains. Soil was also collected from same location. Contents of heavy metals (Cd, Cu, Pb, Zn, Ni, Cr, and As) were determined spectrometically in fruiting bodies of forty eight wild macrofungi specimens with soils. When the analysed their transfer factor from soil to fruiting body, Amanita volvata have the highest transfer factor of cadmium and arsenic. And Mycena pura showed the highest in mercury, Marasmius pulcherripes in zinc, Laccaria laccata in nickel, and Collybia confluens in chrome. When compare the population of mushrooms between the contaminated mines and Mt. Chiak as the healthy area, Genus Russula and Collybia were collected both area, but Leucocoprinus, Coprinus, Suillus, Lepiost, Gyroporus, Lepista, Microstoma, Stropharia, and Agrocybe were only in the contaminated mine area.

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Purification and Characterization of Gibberellin $3Beta$-Hydroxylase from Immature Seeds of Phaseolus vulgaris (강낭콩미숙종자로부터 Gibberellin $3Beta$-Hydroxylase 정제 및 성질)

  • 곽상수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.133-148
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    • 1987
  • Gibberellin(GA) 3-$\beta$ hydroxylation is very important for the shoot elogation in the higher plants, since only 3$\beta$-hydryoxylated GAs promote shoot elogation in several plants. Fluctuation of 3$\beta$-hydryoxylase activity was examined during seed maturation using two cultivars of , P. vulgaris, Kentucky Wonder (normal) and Masterpiece (dwarf). Very immature seeds of both cultivars contain high level of 3$\beta$-hydroxylase activity (per mg protein). Both cultivars showed maximum of enzyme activity (per seed) in the middle of their maturation process. Gibberellin 3$\beta$-hydroxylase catalyzing the hydroxylation of GA20 to GA1 was purified 313-fold from very early immature seeds of P. vulgaris. Crude soluble enzyme extracts were purified by 15% methanol precipitation, hydrophobic interaction chromatogrphy, DEAE ion exchange column chromatography and gel filtration HPLC. The 3$\beta$-hydroxylase activity was unstable and lost much of its activity duting the purification. The molecular weight of purified enzyme was extimated to be 42, 000 by gel filtration HPLC and SDS-PAGE. The enzyme exhibited maximum activity at pH 7.7. The Km values for [2.3-3H] GA20 and [2.3-3H]GA9 were 0.29 $\mu$M and 0.33 $\mu$M, respectively. The enzyme requires 2-oxoglutarate as a cosubstrate; the Km value for 2-oxoglutarate was 250 $\mu$M using 3H GA20 as a substrate. Fe2+ and ascorbate significantly activated the enzyme at all purification steps, while catalase and BSA activated the purified enzyme only. The enzyme was inhibited by divalent cations Mn2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+ and Hg2+. Effects of several GAs and GA anaogues on the putrified 3$\beta$-hydroxylase were examined using [3H]GA9 and GA20 as a substrates. Among them, GA5, GA9, GA15, GA20 and GA44 inhibited the enzyme activity. [13C, 3H] GA20 was converted by the partially purified enzyme preparation to [13C, 3H]GA1, GA5 and GA6, which were identified by GC-MS, GA9 was converted only GA4, GA15 and GA44 were converted to GA37 and GA38, respectively. GA5 was epoxidized to GA6 by the preparation. This suggests that 3$\beta$-hydroxylation of GA20 and epoxidation of GA5 are catalyzed by the same enzyme in P, vulgaris.

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Chemical Components and Physiological Activities of Bamboo (Phyllostachys bambusoides Starf) Extracts Prepared with Different Methods (추출방법에 따른 대나무(왕대) 추출물의 화학성분 및 생리활성)

  • Ju, In-Ok;Jung, Gi-Tai;Ryu, Jeong;Choi, Joung-Sik;Choi, Yeong-Geun
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.542-548
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    • 2005
  • Chemical components and physiological activities Bamboo (Phyllostachys bambusoides Starf) extracts obtained by burning, dry heating or extracting with water or 70% ethanol and were investigated. Contents of soluble solid and total phenolic compounds were highest in the ethanol extract. Contents of polyphenols such as catechin, chlorogenic acid, caffeic acid, 3-hydroxy benzoic acid and ferulic acid were determined. Free sugars consisted of galactose, glucose, fructose, and sucrose. Organic acids including citric, tartaric, malic, succinic, and acetic acid were present in the bamboo extracts. Antioxidant activities of dry heat and ethanol extracts were higher than those of BHA or ${\delta}-tocopherol$. Nitrite- scavenging effect of extracts ranged from 84.7 to 99.6% at pH 1.2 ana 3.0. Tyrosinase-inhibitory activity was higher in the water extract, and SOD-like and ACE-inhibitory activity were highest in tile dry kent extract. Antimicrobial activities of the bamboo extracts were strong against Bacillus subtilis, Escherichia coli O157, and Staphylococcus aureus.

Behavior of heavy metals in the surface waters of the Lake Shihwa and its tributaries (시화호와 주변 하천 표층수중의 중금속 거동 특성)

  • Kim Kyung Tae;Lee Soo Hyung;Kim Eun Soo;Cho Sung Rok;Park Chung Kil
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.5 no.1
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    • pp.51-67
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    • 2002
  • In order to understand behaviors of heavy metals around the artificial Lake Shihwa in the vicinity of Kyunggi Bay in Korea in relation with huge environmental changes due to construction of huge artificial lake, water samples were collected from Lake Shihwa and its tributaries from 1996 to 1998 and analyzed. Due to extreme pollutant discharge from various kinds of anthropogenic sources such as the Banweol and Shihwa Industrial Complexes and cities, the Shihwa and its tributaries have been polluted in waters with various heavy metals. The enrichment factors of particulate heavy metals in water of streams and storm sewers were very high. All of the heavy metals observed in the waters showed relatively high temporal and spatial variations. In surface waters of the lake during the desalination after the dike establishment, spatial distributions of heavy metal concentrations were mainly controlled by various biogeochemical factors as well as input of industrial and municipal wastewaters, while, physical mixing was minor factor Pb and Co showed a strong affinity to particle phase, however the affinity to dissolved phase was dominated in Ni, Cu and Cd. Water quality of the artificial Lake Shihwa has been deteriorated by direct discharge of untreated wastewater and heavy metals have been accumulated in the lake system. Therefore, luther environmental improvement plan should be programmed subsequently.

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필름 스피커 적용을 위한 PZT/polymer 복합체의 후막 제조 및 압전 특성 평가

  • Son, Yong-Ho;Eo, Sun-Cheol;Kim, Seong-Jin;Gwon, Seong-Yeol;Gwon, Sun-Yong
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.11a
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    • pp.346-346
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    • 2007
  • 압전세라믹 재료는 현재 압전 변압기, actuator, transducer, sensor, speaker 등에 광범위하게 이용이 되고 있다. 이 중에서 압전세라믹 소결체를 이용한 스피커의 제조는 가공이 까다롭고, 대형의 크기로 제작 시 소자가 깨지는 등의 많은 제약을 받고 있으며, 저음 특성이 떨어져 응용 범위가 한정되어 있다. 따라서 최근에는 이러한 단점을 극복하기 위하여 세라믹/고분자 복합체를 이용한 필름 스피커를 제작하고자 시도하고 있다. 이러한 세라믹/고분자 0-3형 압전 복합체를 이용할 경우, 제품의 경량화를 실현할 수 있고, 크기나 환경의 영향을 거의 받지 않으므로, 고기능성 스피커로의 응용에 적합할 것으로 보인다. 따라서 본 연구에서는 PZT계의 세라믹와 PVDF, PVDF-TrFE, Polyester, acrylic resin 등의 여러 고분자 물질과의 복합체를 제조하여 압전특성을 평가하였다. 본 실험은 먼저 $(Pb_{1-a-b}Ba_aCd_b)(Zr_xTi_{1-x})_{1-c-d}(Ni_{1/3}Nb_{2/3})_c(Zn_{1/3}Nb_{2/3})_dO_3$ (이하 PZT라 표기)의 최적화 조성을 선택하여, $1050^{\circ}C$에서 소결된 분말을 48시간 ball milling방법 로 약 $1{\mu}m$ 크기로 분쇄하였다. 고분자 물질들은 알맞은 용제들을 선택하여 녹였다. 그 다음 소결된 PZT분말과 고분자를 50:50, 60:40, 65:35, 70:30등의 무게 분율로 혼합하고, 분산제, 소포제 등을 첨가하여 3단 roll mill을 이용하여 충분히 분산시켜 페이스트 (Paste)를 제조하였다. 제조된 페이스트를 ITO가 코팅된 PET필름 위에 스크린 프린팅 법을 사용하여 인쇄하여 $120^{\circ}C$에서 5분간 건조하였다. 코팅된 복합체의 두께는 약 $80{\mu}m$ 정도로 측정되었다. Ag 페이스트를 이용한 상부 전극 형성에도 스크린 프린팅 법을 적용하였다. 이를 $120^{\circ}C$에서 4 kV/mm의 DC 전계로 분극 공정을 수행한 후 전기적 특성을 평가하였다. 유전특성을 조사하기 위해서 LCR meter (EDC-1620)를 사용하였고, 시편의 결정구조는 XRD (Rigaku; D/MAX-2500H)을 통해 분석하였으며, 전자현미경(SEM)을 이용하여 미세구조를 분석하였다. 압전 전하상수$(d_{33})$ 값은 APC 8000 모델을 이용하여 측정하였다. PZT의 혼합비가 증가할수록 비유전율 및 압전 전하 상수 등의 전기적 특성이 증가되었다. 또 여러 고분자 물질 중에서 PVDF-TrFE 수지가 가장 우수한 특성을 보였다. 이는 PVDF-TrFE 수지가 압전성을 나타내기 때문인 것으로 판단되었다.

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다층 PNN-PZT/Ag 복합체의 동시 소성을 위한 압전세라믹스의 저온소결 및 압전특성 평가

  • Lee, Myeong-U;Son, Yong-Ho;Kim, Seong-Jin;Yun, Man-Sun;Ryu, Seong-Rim;Gwon, Sun-Yong
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.11a
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    • pp.295-295
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    • 2007
  • 기계적 에너지를 전기적 에너지로 변화하는 에너지 변환소자인 압전 세라믹스는 액츄에이터, 변압기, 초음파모터, 초음파 소자 및 각종 센서로 응용되고 있으며, 그 응용분야는 크게 증가하고 있다. 최근 이러한 에너지 변화 소자는 앞으로 도래하는 ubiquitous, 무선 모바일 시대의 휴대용 전자제품, robotics, 항공우주, 자동차, 의료, 건축, MEMS 분야 등의 대체 에너지원으로 응용하기 위한 연구가 진행되고 있다. 특히 인간의 동작 등과 같은 일상적인 동작으로 필요한 전력을 얻을 수 있고, 세라믹 소자를 이용하기 때문에 전자노이즈가 발생되지 않을 뿐 아니라 반영구적으로 사용할 수가 있어서, 기존 이차전지, 연료전지를 대체 또는 보완 할 수 있는 방안도 검토되고 있다. PZT계 세라믹스는 높은 유전상수와 압전특성으로 전자세라믹스분야에서 가장 널리 사용되어지고 있지만 $1200^{\circ}C$이상의 높은 소결온도 때문에 $1000^{\circ}C$ 부근에서 급격히 휘발되는 PbO로 인한 환경오염과 기본조성의 변화로 인한 압전 특성의 저하가 문제시되고 있다. 또한, 적층 세라믹스의 제작 시 구조적 특성상 내부 전극이 도포된 상태에서 동시 소결이 필요한데, 융점이 낮은 Ag전극 대신 값비싼 Pd나 Pt가 다량 함유된 Ag/Pd, Ag/Pt 전극이 사용되고 있어 경제적인 문제가 발생하게 된다. 따라서 순수 Ag 전극을 사용하거나, Ag의 비율이 높은 내부 전극을 사용하기 위해서는 $950^{\circ}C$ 이하에서 소결되는 압전 세라믹스를 개발 하는 것이 필요하다. 따라서 본 연구에서는 압전특성이 우수한 $(Pb_{1-x}Cd_x)\;[(Ni_{1/3}/Nb_{2/3})_{0.25}Zr_{0.35}Ti_{0.4}]O_3$계의 조성을 설계하여, 소결온도를 낮추기 위해서 2단계 하소법을 이용하였다. 분말을 ball milling을 통해 24시간 동안 혼합하였다. 혼합된 분말은 $800^{\circ}C$에서 2시간 동안 하소하였다. 하소한 분말을 72시간 동안 ball milling 하여 최종 분말을 얻었다. 최종 분말에 PVB를 첨가하여 ${\Phi}21$ disk 형태로 성형한 후, $800{\sim}950^{\circ}C$ 소결을 하였다. 최종 분말 및 소결된 시편을 XRD분석을 통하여 상을 확인하였고, SEM을 이용하여 미세조직을 관찰하였다. 전기적 특성을 확인하기 위하여 두께 1mm로 연마한 시편에 Ag 전극을 도포하여 열처리한 후, 분극 처리하였다. 압전특성은 $d_{33}$ 미터로 측정하였고, impedance analyzer를 이용하여 주파수 및 impedance 특성을 측정하였다. 그 결과 $900^{\circ}C$에서 우수한 압전 특성 및 전기적 특성을 확보 할 수 있었다.

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Physical and Chemical Properties of Cover Soils of waste Landfills in Kyonggi-Do Area (경기도 지역 쓰레기 매립지 복토층 토양의 이화학성)

  • 이상모;김기대;이은주;김판기;이군택
    • Journal of Soil and Groundwater Environment
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    • v.7 no.1
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    • pp.53-62
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    • 2002
  • The physical and chemical properties of cover soils of 10 waste landfill sites in Kyonggi-Do area, where social circumstances at present forces to consider the reuse of landfill, were investigated to provide the informations of soil environment which are necessary to establish the appropriate ecological restoration plan of waste landfills. The pH and electrical conductivity of soils were higher in landfills sites than in reference sites (area around landfill sites), indicating the salt accumulation in surface soil. However, total-N and organic matter contents were lower in landfills sites than in reference sites. In landfill sites, the total-N and plant available-P contents were less than 0.15% and 20mg/kg, respectively. Exchangeable cations (K, Ca, Mg and Na) and heavy metal (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) contents varied between the landfill sites, but were higher in landfills sites than in reference sites. The major exchangeable cation of soil was Ca. Heavy metal contents were much lower than the critical concentration which phytotoxicity is considered to be possible and the standard for agricultural land of Korean Soil Environmental Preservation Act. Therefore, the proper soil management plan to increase the soil fertility is recommended for the ecological restoration of landfill using natural or artificial vegetation.

Behavior Changes of Earthworm from Soils Amended with Biochar - Avoidance and Productivity - (바이오차르 토양 투입에 따른 지렁이의 행동변화 연구 - 회피 및 생산성 변화 -)

  • Kim, You Jin;Yang, Seung Hoon;Kim, Seo Yeon;Yoon, Hong Seok;Yoo, Ga Young
    • Journal of Climate Change Research
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    • v.5 no.4
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    • pp.277-284
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    • 2014
  • Biochar application to soil is widely known to have effects of climate change mitigation and soil quality improvement. However, effects of biochar on soil ecosystem are not always positive and some biochars are reported to contain toxic materials which might influence soil ecosystem. In this context, this study aims to investigate behavioral changes of earthworms(Eisenia fetida, Eisenia andrei) in response to different application rates of biochar to artificial soil. Treatment included two types of biochars made from rice husk (RH_Char) and wastewater sludge (SL_Char) with 1% and 10% application rates, respectively. Avoidance test revealed that earthworms did not avoid SL_Char treatments at 1% and 10%, while they rather moved to the RH_Char treatments probably due to higher labile carbon content(Hot water extractable carbon) of the RH_Char. The HWC content of RH_Char was 4 times higher than that of the SL_Char. Results of reproduction test showed that the survival rates, number of juveniles and number of cocoons were not influenced by biochar application except for the treatment of SL_Char at 10% rate. In the SL_Char 10% treatment, fatality was approximately 3.3 times as high as the control and the number of cocoons was 1.3 times higher in the same treatment than the control, indicating that earthworms were under environmental stress. The possible explanation for the stress condition was related to higher Cd, Ni, Cr, and As contents in the SL_Char. Overall results imply that biochar application at low rate might not change earthworms' behavior for the short term, while the reproduction behavior might be negatively influenced under the high application rate.