• Title/Summary/Keyword: P-E

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Comparative Study on Characterization of Recombinant B Subunit of E. coli Heat-Labile Enterotoxin (rLTB) Prepared from E. coli and P. pastoris

  • Ma, Xingyuan;Yao, Bi;Zheng, Wenyun;Li, Linfeng
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.550-557
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    • 2010
  • Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6$\times$His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at $30^{\circ}C$, whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at $37^{\circ}C$. The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6$\times$His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6$\times$His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.

Effects of Intraperitoneally Administered Lipoic Acid, Vitamin E, and Linalool on the Level of Total Lipid and Fatty Acids in Guinea Pig Brain with Oxidative Stress Induced by H2O2

  • Celik, Sait;Ozkaya, Ahmet
    • BMB Reports
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    • v.35 no.6
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    • pp.547-552
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    • 2002
  • The aim of our study was to investigate the protective effects of intraperitoneally-administrated vitamin E, dl-alpha lipoic acid, and linalool on the level of total lipid and fatty acid in guinea pig brains with oxidative stress that was induced by $H_2O_2$. The total brain lipid content in the $H_2O_2$ group decreased when compared to the $H_2O_2$ + vitamin E (p<0.05), $H_2O_2$ + linalool (p<0.05), ALA (p<0.05), control (p<0.01), linalool (p<0.01), and vitamin E (p<0.01) groups. While the proportion of total saturated fatty acid (${\Sigma}SFA$) in the $H_2O_2$ group significantly increased (p<0.005) when compared to the vitamin E group, it only slightly increased (p<0.01) when compared to the control and $H_2O_2$ + vitamin E groups. The ratio of the total unsaturated fatty acid (${\Sigma}USFA$) in the $H_2O_2$ groups was lower (p<0.05) than the control, vitamin E, and $H_2O_2$ + vitamin E groups. The level of the total polyunsaturated fatty acid (${\Sigma}PUEA$) in the $H_2O_2$ group decreased in when compared to the control, vitamin E, and $H_2O_2$ + vitamin E groups. While the proportion of the total w3 (omega 3), w6 (omega 6), and PUFA were found to be lowest in the $H_2O_2$ group, they were slightly increased (p<0.05) in the lipoic acid group when compared to the control and $H_2O_2$ + lipoic acid groups. However, the level of ${\Sigma}SFA$ in the $H_2O_2$ group was highest; the level of ${\Sigma}USFA$ in same group was lowest. As the proportion of ${\Sigma}USFA$ and ${\Sigma}PUFA$ were found to be highest in the linalool group, they were decreased in the $H_2O_2$ group when compared to the control group. Our results show that linalool has antioxidant properties, much the same as vitamin E and lipoic acid, to prevent lipid peroxidation. Additionally, vitamin E, lipoic acid, and linalool could lead to therapeutic approaches for limiting damage from oxidation reaction in unsaturated fatty acids, as well as for complementing existing therapy for the treatment of complications of oxidative damage.

타원편광분석법을 이용한 $In_xAl_{1-x}P$ 박막의 광물성 연구

  • Byeon, Jun-Seok;Hwang, Sun-Yong;Kim, Tae-Jung;Kim, Yeong-Dong;Aspnes, D.E.;Chang, Y.C.;Yun, Jae-Jin;Lee, Eun-Hye;Bae, Min-Hwan;Song, Jin-Dong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.423-423
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    • 2013
  • 3~5 족 반도체 물질인 phosphorus 화합물 중 대표적인 InAlP 삼종화합물은 작은 굴절률, 큰 밴드갭, GaAs와 lattice 일치 때문에 큰 주목을 받고 있고, p-type high electron mobility transistors(p-HEMT), laser diodes 등의 고속 전자소자 및 광전 소자에 응용이 가능한 매우 중요한 물질이다. 최적의 소자 응용기술을 위해서는, 정확한 광물성 연구가 수행되어야 하지만 InxAl1-xP 화합물에 대한 유전율 함수 및 전자전이점 등의 연구는 미흡한 실정이다. 이에 본 연구에서는 1.5~6.0 eV 에너지 영역에서 각기 다른 In 조성비를 갖는 InxAl1-xP 화합물의 가유전율 함수 ${\varepsilon}={\varepsilon}_1+i{\varepsilon}_2$와 전자전이점 데이터를 보고한다. GaAs 기판 위에 molecular beam epitaxy (MBE)를 이용하여 InxAl1-xP (x=0.000, 0.186, 0.310, 0.475, 0.715, 0.831, 1.000) 박막을 성장하였고 타원편광분석기를 이용하여 유전율 함수를 측정하였다. 또한 실시간 화학적 에칭을 통하여 시료 표면에 자연산화막을 제거함으로써 순수한 InAlP의 유전율 함수를 측정할 수 있었고, 측정된 유전율 함수를 이차미분하여 In 조성비에 따른 전자전이점을 얻을 수 있었다. 얻어진 전자전이점 값을 이용하여 linear augmented Slater-type orbital method (LASTO) 를 통해 이론적 전자 밴드 구조 계산을 하였고, 이를 바탕으로 $E_0$, $E_1$, $E_2$ 전이점 지역의 여러 전자전이점($E_1$, $E_1+{\Delta}_1$, $E_0'$, $E_0'+{\Delta}_0'$, $E_2$, $E_2'$)의 특성을 정의할 수 있었고, $E_0'$$E_2$ 전이점의 에너지 값이 In 조성비가 증가함에 따라 서로 교차함을 발견할 수 있었다. 타원 편광 분석법을 이용한 유전율 함수 및 전자전이점 연구는 InAlP의 광학적 데이터베이스를 확보하는 성과와 더불어 새로운 디바이스 기술 및 광통신 산업에도 유용한 정보가 될 것이다.

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Purification and Characterization of the Overproduced E. coli Endochitinase (과량 생산된 대장균 chitin 분해효소의 정제 및 특성 조사)

  • Hwang, Hee-Young;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.171-175
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    • 2003
  • The putative endochitinase gene, yheB of Escherichia coli K-12 is not expressed under lab culture conditions. The endochitinase gene was amplified by PCR and subcloned into pET28c vector and pQE9 vector, respectively. The endochitinase produced in E. coli harboring pET28c containing yheB or pQE9 vector containing yheE was partly released into the growth medium. The overproduced endochitinase was partially purified by His affinity column chromatography and DE-52 column chromatography. The apparent molecular weight of the endochitinase determined by SDS-polyacrylamide gel electrophoresis was about 97,000. The purified E. coli endochitinase showed maximal chitinolytic activity at pH 6 and $40^{\circ}C$.

Effect of Dietary Vitamin E on the Microsomal Mixed Function Oxidase System of Liver and Lung in Streptozotocin-induced Diabetic Rats (식이 Vitamin E가 Streptozotocin 유발 당뇨쥐의 간 및 폐조직에서의 Microsomal Mixed Function Oxidase System에 미치는 영향)

  • 박영란;이순재;임영숙;주길재
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.6
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    • pp.969-975
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    • 1996
  • The purpose of this study was to investigate the effect of dietary vitamin E on microsomal mixed function oxidase system of liver and lung in streptozotocin(STZ) induced diabetic rats. Sprague-Dawley male rats weighing 140 $\pm$ 10mg were randomly assigned to one control and three STZ-diabetic groups. Diabetic groups were divided into DM-0E(vitamin E free diet), DM-40E(40mg vitamin E kg/diet) and DM-400E(400mg vitamin E kg/diet) according to the level of vitamin E supplementation. Diabetes was experimentally induced by intravenous administration of 55mg/kg b.w of STZ in citrate buffer(pH 4.3) after 4 week feeding of three experimental diets. Animals were sacrificed at the 6th day of diabetic state. The contents of cytochrome P$_{450}$ in DM-0E, DM-40E and DM-400E groups of liver were increased by 162%, 150% and 56%, respectively, compared with that of control. Also the contents of cytochrome P$_{450}$ in lung were similar to liver. The activities of cytochrome bs in DM-0E and DM-40E groups of the liver were increased by 70% and 53%, respectively, compared with that of control, but not in DM-400E group. The activities of bs in DM-0E, DM-40E and DM-400E groups of lung were signficantly increased. Activity of cytochrome P$_{450}$ reductase in DM-0E, DM-40E of liver and lung were higher than that of control group, but the activity of DM-400E group was not different from that of control. The lipid peroxide values of DM-0E, DM-40E and DM-400E groups were 143%, 95% and 31% higher than those of control. It was concluded that dietary vitamin E had protective effects on lipid peroxidation accompanied with increased mixed function of oxidase activity in diabetic rats.

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INJECTIVE AND PROJECTIVE PROPERTIES OF REPRESENTATIONS OF QUIVERS WITH n EDGES

  • Park, Sangwon
    • Korean Journal of Mathematics
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    • v.16 no.3
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    • pp.323-334
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    • 2008
  • We define injective and projective representations of quivers with two vertices with n arrows. In the representation of quivers we denote n edges between two vertices as ${\Rightarrow}$ and n maps as $f_1{\sim}f_n$, and $E{\oplus}E{\oplus}{\cdots}{\oplus}E$ (n times) as ${\oplus}_nE$. We show that if E is an injective left R-module, then $${\oplus}_nE{\Longrightarrow[50]^{p_1{\sim}p_n}}E$$ is an injective representation of $Q={\bullet}{\Rightarrow}{\bullet}$ where $p_i(a_1,a_2,{\cdots},a_n)=a_i,\;i{\in}\{1,2,{\cdots},n\}$. Dually we show that if $M_1{\Longrightarrow[50]^{f_1{\sim}f_n}}M_2$ is an injective representation of a quiver $Q={\bullet}{\Rightarrow}{\bullet}$ then $M_1$ and $M_2$ are injective left R-modules. We also show that if P is a projective left R-module, then $$P\Longrightarrow[50]^{i_1{\sim}i_n}{\oplus}_nP$$ is a projective representation of $Q={\bullet}{\Rightarrow}{\bullet}$ where $i_k$ is the kth injection. And if $M_1\Longrightarrow[50]^{f_1{\sim}f_n}M_2$ is an projective representation of a quiver $Q={\bullet}{\Rightarrow}{\bullet}$ then $M_1$ and $M_2$ are projective left R-modules.

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Exhaled Nitric Oxide(NO) Among Adult Male Workers (성인남성 근로자들의 호기중 일산화질소)

  • 하태규;백도명
    • Journal of Environmental Health Sciences
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    • v.29 no.3
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    • pp.35-42
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    • 2003
  • Nitric Oxide(NO) is produced in many organs of the body, including the lung and airways, and it is detectable in the exhaled air. The measurement of exhaled NO(eNO) provides a simple non-invasive means for measuring airway inflammation, such as asthma. We measured eNO among adult male workers to examine the distribution of eNO in healthy people and to find factors affecting eNO. We measured eNO in a sample of 921 adult workers who also performed lung function test and skin prick test. Exhaled NO was measured in a sitting posture without using a nose clip and NO free gas. NO was measured at three expiratory rates(l8; 42; 71 $m\ell$/sec) and the flow rate of 71 $m\ell$/sec was used in analysis. The average eNO concentration was 5.29 $\pm$ 2.98 ppb. The level increased with age but not significantly(P=0.0529). Exhaled NO showed positive relations to the height(P=0.0001), pollen 1 (P=0.0124), asthma history(P=0.0212), allergic rhinitis symptom(P=0.0302). Exhaled NO Concentration of smokers( 4.62 ppb) was significantly lower than that of nonsmokers(5.99 ppb; P<0.0001).

Antagonistic Action of Lactobacilli Toward Pathogenic Bacteria in Associative Cultures (Lactobacillus spp.에 의한 병원성 세균의 생육저해)

  • 강국희;성문희
    • Journal of Food Hygiene and Safety
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    • v.4 no.2
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    • pp.155-163
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    • 1989
  • Three species of lactobacilli (L. casei, L. acidophilus, L. bulgaricus) were tested for their antibacterial activity. They all were antagonistic to growth of enteropathogenic Escherichia coli and Salmonella enteritidis in associative cultures in YS-medium (0.1 % yeast extract + skimmilk). Sal. enteritidis was more sensitive to the inhibition than was E. coli. Control cultures of E. coli and Sal. enteritidis were pH 5.08 and 5.70 in 72 hrs of incubation and the associative cultures were pH 3.35-4.48. The increases in pH resulting from growth of the lactobacilli in the associative cultures appeared to be sufficient and mainly responsible for the antagonistic actions exerted on the pathogens.

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Characterization of promotor sequences for strong expression of groEx IN Escherichia coli.

  • Lee, Jung E.;Lim, Ssang T.;Ahn, Tae I.
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.15-22
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    • 1996
  • The cloned X-bacterial gene (groEx) which is analogous to groE of E. Coli strongly expressed in E. coli when grown at the temperature 27.deg. C or higher without having to add any inducers. By S1-nuclease mapping, primer extension analysis and site directed mutagenesis, we found 4 promoters in the gene. Among them two promoters located at 5'-extended region of the gene are homologous to the promoters found in groE family of heat-shock genes ; they are , .sigma.$^{32}$ factor-dependent P1 promotor and .delta$^{70}$factor-dependet P2 promoter. The other two promoters found within the coding region of groESx were P3, 5'-TTGGCG-(18 bases)-AATACT-3' and P4, 5'-TTGGCA-(19 bases)-TAAGT which overlapped within 49 bases. These unique intragenic .delta.$^{70}$-dependent promoters are the first to be cloned and characterized in groE analogous heat-shock genes so far. These P3 and P4 promoters appeared to be responsible for the strong expression of GroElx in X-bacteria in vivo.

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Investigation of Endurance Degradation in a CTF NOR Array Using Charge Pumping Methods

  • An, Ho-Myoung;Kim, Byungcheul
    • Transactions on Electrical and Electronic Materials
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    • v.17 no.1
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    • pp.25-28
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    • 2016
  • We investigate the effect of interface states on the endurance of a charge trap flash (CTF) NOR array using charge pumping methods. The endurance test was completed from one cell selected randomly from 128 bit cells, where the memory window value after 102 program/erase (P/E) cycles decreased slightly from 2.2 V to 1.7 V. However, the memory window closure abruptly accelerated after 103 P/E cycles or more (i.e. 0.97 V or 0.7 V) due to a degraded programming speed. On the other hand, the interface trap density (Nit) gradually increased from 3.13×1011 cm−2 for the initial state to 4×1012 cm−2 for 102 P/E cycles. Over 103 P/E cycles, the Nit increased dramatically from 5.51×1012 cm−2 for 103 P/E cycles to 5.79×1012 cm−2 for 104 P/E cycles due to tunnel oxide damages. These results show good correlation between the interface traps and endurance degradation of CTF devices in actual flash cell arrays.