• 제목/요약/키워드: Soluble factor

검색결과 383건 처리시간 0.025초

Soluble Expression and Purification of the Catalytic Domain of Human Vascular Endothelial Growth Factor Receptor 2 in Escherichia coli

  • Wei, Jia;Cao, Xiaodan;Zhou, Shengmin;Chen, Chao;Yu, Haijun;Zhou, Yao;Wang, Ping
    • Journal of Microbiology and Biotechnology
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    • 제25권8호
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    • pp.1227-1233
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    • 2015
  • Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis through binding to its specific receptors, which mainly occurs to VEGF receptor 2 (VEGFR-2), a kinase insert domain-containing receptor. Therefore, the disruption of VEGFR-2 signaling provides a promising therapeutic approach for the treatment of cancer by inhibiting abnormal or tumorinduced angiogenesis. To explore this potential, we expressed the catalytic domain of VEGFR-2 (VEGFR-2-CD) as a soluble active kinase in Escherichia coli. The recombinant protein was purified and the VEGFR-2-CD activity was investigated. The obtained VEGFR-2-CD showed autophosphorylation activity and phosphate transfer activity comparable to the commercial enzyme. Furthermore, the IC50 value of known VEGFR-2 inhibitor was determined using the purified VEGFR-2-CD. These results indicated a possibility for functional and economical VEGFR-2-CD expression in E. coli to use for inhibitor screening.

Cytotoxicity of Root Canal Sealers Containing Calcium Hydroxide

  • Kim, In-Geol;Han, Se-Hee;Cho, Young-Gon;Lee, Sang-Bae;Kim, Kwang-Mahn;Kim, Kyoung-Nam
    • Journal of Korean Dental Science
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    • 제2권1호
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    • pp.11-18
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    • 2009
  • The purpose of this study was to investigate the possibility to reduce the toxicity of oil based root canal sealers containing calcium hydroxide using MTT & agar overlay assays. Thus some formulations of traditional root canal sealers were replaced with oil-soluble solvents and experimental root canal sealers manufactured. In MTT assay, Cell viability of all experimental sealers in addition with oil soluble solvents were observed significantly higher than both control groups, especially according to replace zinc and/or calcium ion components. Also agar overlay assay was appeared moderate to no cell responses into modifying both zinc and/or calcium ion components and oil soluble solvent weight. Authors found the reducing effect of cell toxicity through significant role of oil soluble solvent factor into root canal sealer containing calcium hydroxide.

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지질대사(脂質代謝)에 관여하는 인자(因子) (Some Factors Affecting Lipid Metabolism)

  • 남현근
    • 한국식품영양과학회지
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    • 제15권2호
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    • pp.191-200
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    • 1986
  • It is now generally accepted that individuals at increased risk for cardiovascular disease may be identified by certain traits or habbits. The factors such as high blood pressure, elevated blood cholestrol, age, sex and obesity are associated with increseaed frequency of disease. The blood cholesterol level lowering will decrease cardiovascular disease risk. The regression of atherosclerosis can be achieved by lowering the level of circulating cholesterol. Those things are connected with the quantity and quality of protein, fats, carbohydrates, especially soluble and non-soluble fiber, magnesium and calcium. The lipoprotein and lipid metabolism are connected with the lipid transport. The factors on lipid absorption and blood serum lipid pattern of human are exist. The factors have a variety of materials with different chemical and physical properties. The soluble fiber diet make a low blood and liver lipids. Many kind of soluble fiber results in a lowering of blood cholesterol and triglyceride levels. The cholesterol lowering effects of dietery fiber may be a results of alterations of in intestinal handling of fats, hepatic metabolism of fatty acid or triglyceride acid metabolism of lipoprotein. It is investigated that the high density lipoprotein (HDL) is inversely related to coronary artery disease. It has been postulated that HDL may be an important factor in cholesterol efflux from the tissues, therby reducing the amount of cholesterol deposited there. Alternatively, the HDL may pick up cholestyl ester and phospholipid during normal VLDL lipolysis in the plasma. The HDL levels are relatively insensitive to diet. At present time, the cause-and -diet effect of HDL's inverse relation to CHD remains unclear.

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돼지 간중의 Monooxygenase 가 Diazinon 의 분해에 미치는 영향 (The degradation o Diazinon by hepatic monooxygenase of Pig)

  • 류종국;이규승
    • 한국환경농학회지
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    • 제9권2호
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    • pp.153-159
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    • 1990
  • 돼지간을 두 fraction($105,000{\time}g$ pellet, superatant)으로 나누어 Diazinon의 분해에 관여하는 두 효소(Monooxygenase와 esterase)의 활성 및 저해제(EPN, Beam, PBO)첨가에 따른 Diazinon의 분해억제 능력을 비교 검토하였다. 1. 돼지간의 microsomal fraction과 soluble fraction에서 M. O.와 β-esterase 활성은 20분과 60분에 최고활성을 나타내었다. 2. Microsomal fraction에서의 M. O.활성은 NADPH generating system의 첨가로 1.6배의 활성증가와 지속성이 확인되었다. 3. Microsomal fraction에서의 Diazinon분해량은 30분에 최고량(0.060nM/ mg/ min)을 보였으나, soluble fraction에서는 60분에 최고량(0.018nM/ mg/ min)을 보여 1/3수준이었다. 4. M. O.활성에 미치는 Diazinon에 대한 EPN, Beam, PBO의 첨가효과는 Beam(14%)< EPN(15%)

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Soluble Expression of Recombinant Olive Flounder Hepcidin I Using a Novel Secretion Enhancer

  • Lee, Sang Jun;Park, In Suk;Han, Yun Hee;Kim, Young Ok;Reeves, Peter R.
    • Molecules and Cells
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    • 제26권2호
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    • pp.140-145
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    • 2008
  • Expression of olive flounder hepcidin I (HepI) fused with truncated OmpA signal peptides ($OmpASP_{tr}$) as directional signals does not produce soluble fusion proteins. However, by inserting amino acid segments (xxx) varying in pI and hydrophobicity/hydrophilicity into a leader sequence containing a truncated OmpASP ($OmpASP_{tr}$) and a factor Xa cleavage site (Xa) [$OmpASP_{tr}{\mid}(xxx){\mid}Xa$], we were able in some cases to express soluble recombinant HepI. Soluble expression of the recombinant protein strongly correlated with (xxx) insertions of high pI and hydrophilicity. Therefore, we modified the $OmpASP_{tr}{\mid}(xxx){\mid}Xa$ sequence by inserting Arg and Lys into (xxx) to increase the hydrophilicity of the signal peptide region. These modifications enhanced the expression of soluble recombinant HepI. Hydropathic profile analysis of the $OmpASP_{tr}{\mid}(xxx){\mid}Xa$ HepI fusion proteins revealed that the transmembrane-like domains derived from the $OmpASP_{tr}{\mid}(xxx){\mid}Xa$ sequence were larger than the internal positively charged domain native to HepI. It should therefore be possible to overcome the obstacle of internal positively charged domains to obtain soluble expression of recombinant proteins by monitoring the hydrophilicity and hydropathic profile of the signal peptide region using a computer program.

The Effect of Growth Condition on a Soluble Expression of Anti-EGFRvIII Single-chain Antibody in Escherichia coli NiCo21(DE3)

  • Dewi, Kartika Sari;Utami, Ratna Annisa;Hariyatun, Hariyatun;Pratiwi, Riyona Desvy;Agustiyanti, Dian Fitria;Fuad, Asrul Muhamad
    • 한국미생물·생명공학회지
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    • 제49권2호
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    • pp.148-156
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    • 2021
  • Single-chain antibodies against epidermal growth factor receptor variant III (EGFRvIII) are potentially promising agents for developing antibody-based cancer treatment strategies. We described in our previous study the successful expression of an anti-EGFRvIII scFv antibody in Escherichia coli. However, we could also observe the formation of insoluble aggregates in the periplasmic space, limiting the production yield of the active product. In the present study, we investigated the mechanisms by which growth conditions could affect the expression of the soluble anti-EGFRvIII scFv antibody in small-scale E. coli NiCo21(DE3) cultures, attempting to maximize production. The secreted scFv molecules were purified using Ni-NTA magnetic beads and protein characterization was performed using SDS-PAGE and western blot analyses. We used the ImageJ software for protein quantification and determined the antigen-binding activity of the scFv antibody against the EGFRvIII protein. Our results showed that the highest percentage of soluble scFv expression could be achieved under culture conditions that combined low IPTG concentration (0.1 mM), low growth temperature (18℃), and large culture dish surface area. We found moderate-yield soluble scFv production in the culture medium after lactose-mediated induction, which was also beneficial for downstream protein processing. These findings were confirmed by conducting western blot analysis, indicating that the soluble, approximately 30-kDa scFv molecule was localized in the periplasm and the extracellular space. Moreover, the antigen-binding assay confirmed the scFv affinity against the EGFRvIII antigen. In conclusion, our study reveals that low-speed protein expression is preferable to obtain more soluble anti-EGFRvIII scFv protein in an E. coli expression system.

된장의 지용성ㆍ수용성추출물에 대한 갈색 특성 및 항산화 효과 (Brown Color Characteristics and Antioxidizing Activity of Doenjang Extracts)

  • 김현정;손경희;채선희;곽동경;임성경
    • 한국식품조리과학회지
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    • 제18권6호
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    • pp.644-654
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    • 2002
  • 본 연구는 간장을 걸러내지 않고 담은 된장과 메주를 45일 침지 후 간장을 걸러내고 담은 된장을 각각 오지항아리와 투명유리병에서 60, 120, 180일 숙성시키면서, 된장 지용성 및 수용성추출물의 갈색특성 및 항산화 효과를 알아보았다. 된장 지용성 추출물과 수용성 추출물을 각각 제조하여 420 nm에서 갈색도를 측정한 결과, 수용성 추출물이 지용성 추출물보다 갈색도가 높은 것으로 나타났으며 전체적으로 간장을 걸러내지 않고 유리병에 담근 된장이 가장 높은 값을 가졌다. 또한 수용성 추출물의 경우는 숙성기간에 따라 갈색도가 뚜렷이 증가하는 경향을 보였으나 지용성 추출물은 그렇지 못했다. UV-VIS scanning spectra를 측정한 결과, 지용성 추출물은 235nm 부근에서 최대 파장을 보이며 숙성기간에 따른 변화는 거의 없는 것으로 나타났다. 수용성 추출물은 235nm, 340~410nm 부근에서 높은 파장을 나타내었고 숙성기간에 따른 변화를 뚜렷이 보여주었다. Antioxidant activity 실험결과, 지용성 추출물은 숙성기간이 길어짐에 따라 약간 증가하였으나 수용성 추출물은 크게 증가하였다. 환원력과 전자공여능 실험결과 또한 수용성 추출물이 지용성 추출물보다 높은 값을 나타내었으며, 숙성기간이 길어질수록 유리병보다 항아리에 담근 된장이 높은 항산화 효과를 가진 것으로 나타났다. 본 실험결과 전체적으로 숙성 60일에는 간장을 우려낸 된장보다 우려내지 않고 담은 된장에서 아미노산, 환원당, 갈색도, 항산화 효과가 높은 것으로 나타났으나, 숙성기간이 길어질수록 담금용기에 따른 영향을 많이 받아 유리병에 담근 된장보다 항아리에 담근 된장이 아미노산함량과 항산화 효과가 높은 것으로 나타났다.he experiment result shows that our algorithm achieves much better performance.정을 보였으며 5%희석군의 경우 육색은 10일 경과 시점, 냄새는 7일 경과시점부터 초기 부패과정을 보였다. 이상의 결과에서 보는 바와 같이 생체수 파이-워터는 닭고기의 신선도 유지와 관련하여 지방산패를 억제하고 일반세균의 증식을 막아줌으로써 제품의 신선도를 연장시킬 수 있으며 희석농도의 차이에 따른 유의차가 있음을 알 수 있었다.rid형 복합레진이 micro-hybrid형 복합레진에 비해 C-factor의 영향을 더 많이 받는 것으로 나타났으며, 와동의 C-factor증가가 굴곡강도나 탄성계수와 같은 복합레진의 물리적 성질을 저하시킨다는 것을 의미하였다.중합수축. 중합도 및 미세경도 측정치는 one-way ANO-VA와 Duncan's multiple range test를, 변연누출 정도는 chi-square test를 이용, 통계처리하여 다음의 결과를 얻었다. . 중합수축의 정도는 VIP$^{TM}$(Bisco) 사용군에서 전체 조사광도가 높을수록 큰 경향을 보여 600mW/$\textrm{cm}^2$군에서 가장 크게 나타났고, 그 다음으로 Pulse-delay군, 400mW/$\textrm{cm}^2$군, 200 mW/$\textrm{cm}^2$군 순이었고, Optilux 501$^{TM}$(Demetron/Kerr) 사용군에서는 Continuous 방식이 Ramp 방식 보다 크게 나타났다. . 중합도와 미세경도 값은 공히, 전체 조사광도가 높을수록 높게 나타났으며 ,

Multivesicular DepoFoam particles for oral delivery of recombinant human epidermal growth factor

  • Li, Hong;An, Jun-Hee;Park, Jeong-Sook;Han, Kun
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.299.1-299.1
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    • 2003
  • Multivesicular DepoFoam technology is best suited for the encapsulation and sustained release of water-soluble drugs. The purpose of the present study was to prepare multivesicular DepoFoam particles and investigated possibility of oral delivery of a peptide, human epidermal growth factor (rhEGF). The multivesicular DepoFoam particles containing rhEGF was prepared by a two step water-in-oil-in-water double emulsification process. (omitted)

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폐쇄성 수면 무호흡이 Soluble Intercellular Adhesion Molecule-1과 Vascular Endothelial Growth Factor에 미치는 영향 (Influence of Obstructive Sleep Apnea on Soluble Intercellular Adhesion Molecule-1 and Vascular Endothelial Growth Factor)

  • 김정표;이상학;권순석;김영균;김관형;송정섭;박성학;문화식
    • Tuberculosis and Respiratory Diseases
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    • 제56권4호
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    • pp.364-373
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    • 2004
  • 연구배경 : 폐쇄성 수면 무호흡 증후군은 고혈압과 심부정맥 및 허혈성 심질환 등의 심혈관질환을 일으키는 위험요소로 알려져 있으나 정확한 기전에 대해서는 아직 확실히 밝혀져 있지 않다. 최근에 soluble intercellular adhesion molecule-1(sICAM-1)과 vascular endothelial growth factor(VEGF)가 죽상경화증의 발생과 진행에 관여한다고 알려지면서, 일부 연구에서는 이러한 사이토카인이 폐쇄성 수면 무호흡 증후군에서도 증가한다고 보고하였다. 저자는 폐쇄성 수면 무호흡 증후군 환자에서 sICAM-1과 VEGF의 농도 변화를 관찰함으로써, 이들 환자에서 동반될 수 있는 심혈관질환 발생의 병태생리를 일부 이해하고자 하였다. 방 법 : 폐쇄성 수면 무호흡 증후군을 진단하기 위해 가톨릭대학교 성바오로병원 수면장애크리닉을 방문하여 수면다원검사를 시행한 37명을 대상으로 하였다. 철야 수면다원검사를 종료한 직후 혈액 채취를 한 후 혈청분리를 하여 $-70^{\circ}C$ 냉장고에 보관하였다. 사이토카인의 측정은 ELISA 방법으로 시행하였고 수면다원검사의 각 지표와 혈청 sICAM-1 농도 및 혈청 VEGF 농도와의 상관 관계를 비교 분석하였다. 결 과 : 무호흡-저호흡지수와 혈청 sICAM-1 농도 사이에는 유의한 상관관계가 없었지만(r=0.27, P>0.05), 무호흡-저호흡지수와 혈청 VEGF 농도는 유의한 상관관계가 있었다(r=0.50, P<0.01). 무호흡지수와 혈청 sICAM-1 농도는 다소 유의한 상관관계가 있었으며(r=0.31, P<0.01) 무호흡지수와 혈청 VEGF 농도 사이에는 유의한 상관관계가 있었다(r=0.45, P<0.01). 결 론 : 폐쇄성 수면 무호흡 혹은 저호흡은 혈청내 sICAM-1 및 VEGF 농도를 증가시키는 것을 알 수 있었으며, 이러한 사이토카인의 발현 증가는 폐쇄성 수면무호흡 증군 환자에서 심혈관질환의 발생에 관련이 있을 것으로 추측된다.

Molecules of the Tumor Necrosis Factor (TNF) Receptor and Ligand Superfamilies: Endless Stories

  • Kwon, Byung-Suk;Kwon, Byoung-Se
    • BMB Reports
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    • 제32권5호
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    • pp.419-428
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    • 1999
  • Tumor necrosis factor (TNF) receptor members have unique structures composed of 2-4 cysteine - rich pseudorepeats in the extracellular domain. On ligation by trimeric ligand molecules, oligomerization of three receptor molecules occurs, which in turn activates the receptor and recruits intracellular signaling molecules to the cytoplasmic tail to initiate biological events. Recently, the numbers of tumor necrosis factor receptor and ligand family members have been rapidly expanding. Functional characterization of the new members has indicated redundant roles with other known members as well as provided insights into novel functions. In particular, identification of soluble decoy receptors which have the ability to bind multiple ligands highlights a complex control mechanism of immune responses by these molecules. Studies of the new members have also revealed that the TNF receptor and ligand family members play an important role in other than the immune system.

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