• Title/Summary/Keyword: Molecular Surface

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Mitoxantrone Binds to Nopp140, an Intrinsically Unstructured Protein, and Modulate its Interaction with Protein Kinase CK2

  • Lee, Won-Kyu;Lee, Sang-Yeop;Na, Jung-Hyun;Jang, Sung-Woo;Park, Chan-Ryang;Kim, Soo-Youl;Lee, Si-Hyeong;Han, Kyou-Hoon;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.33 no.6
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    • pp.2005-2011
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    • 2012
  • Nopp140 is a highly phosphorylated protein that resides in the nucleolus of mammalian cell and is involved in the biogenesis of the nucleolus. It interacts with a variety of proteins related to the synthesis and assembly of the ribosome. It also can bind to a ubiquitous protein kinase CK2 that mediates cell growth and prevents apoptosis. We found that Nopp140 is an intrinsically unfolded protein (IUP) lacking stable secondary structures over its entire sequence of 709 residues. We discovered that mitoxantrone, an anticancer agent, was able to enhance the interaction between Nopp140 and CK2 and maintain suppressed activity of CK2. Surface plasma resonance studies on different domains of Nopp140 show that the C-terminal region of Nopp140 is responsible for binding with mitoxantrone. Our results present an interesting example where a small chemical compound binds to an intrinsically unfolded protein (IUP) and enhances protein-protein interactions.

The use of HRSEM to characterize new and aged membranes in drinking water production

  • Wyart, Y.;Nitsche, S.;Chaudanson, D.;Glucina, K.;Moulin, P.
    • Membrane and Water Treatment
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    • v.2 no.4
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    • pp.251-266
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    • 2011
  • This work deals with the use of High Resolution Scanning Electron Microscopy (HRSEM) to verify ultrafiltration membrane selectivity at the end of the production line as well as membrane ageing. The first part of this work is focused on new membranes. It is shown that it is better to use sputtering metallization than vacuum deposition, as this latter technique entails thermal damage to the skin layer. Moreover, the impact of the metallization layer on the determination of the membrane pore size is studied and it is observed that no impact of the metallization step can be clearly defined for a metallization layer ranging from 3 to 12 nm. For example, an average pore size of 16.9 nm and a recovery rate of 6.5 % are observed for a 150 kDa cellulose acetate membrane. These results are in agreement with those given by the manufacturer: pore size ranging from 10 to 15 nm and recovery rate ranging from 5 to 10 %. The second part of this work focuses on the study of membrane ageing. A PVDF hollow fibre membrane is studied. It is shown that a 65 % decrease in the permeate flux can be linked to a decrease in the number of pores at the surface of the membrane and a decrease in the recovery rate. In conclusion, a mapping of the pores is performed for several new hollow fibre membranes used to produce drinking water, made of different materials, with different geometries and molecular weight cut-off. These results provide reference data that will help better understand the phenomena of membrane fouling and membrane ageing.

Oleanolic acid 3-acetate, a minor element of ginsenosides, induces apoptotic cell death in ovarian carcinoma and endometrial carcinoma cells via the involvement of a reactive oxygen species-independent mitochondrial pathway

  • Jo, Hantae;Oh, Jeong-Hyun;Park, Dong-Wook;Lee, Changho;Min, Churl K.
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.96-104
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    • 2020
  • Objectives: Oleanolic acid, a minor element of ginsenosides, and its derivatives have been shown to have cytotoxicity against some tumor cells. The impact of cytotoxic effect of oleanolic acid 3-acetate on ovarian cancer SKOV3 cells and endometrial cancer HEC-1A cells were examined both in vivo and in vitro to explore the underlying mechanisms. Methods: Cytotoxic effects of oleanolic acid 3-acetate were assessed by cell viability, phosphatidylserine exposure on the cell surface, mitochondrial release of cytochrome C, nuclear translocation of apoptosis-inducing factor, depolarization of mitochondrial transmembrane potential (∆Ψm), and generation of reactive oxygen species (ROS). In vivo inhibition of tumor growth was also assessed with xenografts in immunocompromised mice. Results: Oleanolic acid 3-acetate exhibited potent cytotoxicity toward SKOV3 and HEC-1A cells by decreasing cell viability in a concentration-dependent manner. Importantly, oleanolic acid 3-acetate effectively suppressed the growth of SKOV3 cell tumor xenografts in immunocompromised mice. Furthermore, oleanolic acid 3-acetate induced apoptotic cell death as revealed by loss of ∆Ψm, release of cytochrome c, and nuclear translocation of apoptosis-inducing factor with a concomitant activation of many proapoptotic cellular components including poly(ADP-ribose) polymerase, Bcl-2, and caspases-8, caspase-3, and caspase-7. Oleanolic acid 3-acetate, however, caused a decrease in ROS production, suggesting the involvement of an ROS-independent pathway in oleanolic acid 3-acetate-induced apoptosis in SKOV3 and HEC-1A cells. Conclusion: These findings support the notion that oleanolic acid 3-acetate could be used as a potent anticancer supplementary agent against ovarian and endometrial cancer. Oleanolic acid 3-acetate exerts its proapoptotic effects through a rather unique molecular mechanism that involves an unconventional ROS-independent but mitochondria-mediated pathway.

Molecular probe for identification of cysts of resting cyst of PSP-producer Alexandrium tamarense (Dinophyceae) (분자생물학적 방법을 이용하여 마비성 패류 독소를 생산하는 알렉산드륨 타마렌스 시스트 탐색)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.13 no.2
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    • pp.163-167
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    • 2003
  • Identification of species within the toxin-producing genus Alexandrium is vital for biotoxin monitoring and mitigation decisions regarding shellfish industry. In particular, the discrimination of resting cysts of only A. tamarense from that of Alexandrium spp. is considerable important to fundamentally monitor and predict this species before vegetative cells occur in the nature. Fluorescent cTAM-F1 DNA probe was responsible to not only binding the activity of the vegetative cells in A. tamarense, but also to the resting cysts, which was treated with methanol after fixation and stained by primuline on the surface The location of fluorescence in cultured vegetative cells and resting cysts was almost at tile bottom of the nucleus. The optimal incubation temperature and time using in situ hybridization were 50-$54^{\circ}C$ and 40-60 min, respectively, to penetrate the DNA probe into cell.

Azasugar-Containing Phosphorothioate Oligonucleotide (AZPSON) DBM-2198 Inhibits Human Immunodeficiency Virus Type 1 (HIV-1) Replication by Blocking HIV-1 gp120 without Affecting the V3 Region

  • Lee, Jinjoo;Byeon, Se Eun;Jung, Ju Yeol;Kang, Myeong-Ho;Park, Yu-Jin;Jung, Kyeong-Eun;Bae, Yong-Soo
    • Molecules and Cells
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    • v.38 no.2
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    • pp.122-129
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    • 2015
  • DBM-2198, a six-membered azasugar nucleotide (6-AZN)-containing phosphorothioate (P = S) oligonucleotide (AZPSON), was described in our previous publication [Lee et al. (2005)] with regard to its antiviral activity against a broad spectrum of HIV-1 variants. This report describes the mechanisms underlying the anti-HIV-1 properties of DBM-2198. The LTR-mediated reporter assay indicated that the anti-HIV-1 activity of DBM-2198 is attributed to an extracellular mode of action rather than intracellular sequence-specific antisense activity. Nevertheless, the antiviral properties of DBM-2198 and other AZPSONs were highly restricted to HIV-1. Unlike other P = S oligonucleotides, DBM-2198 caused no host cell activation upon administration to cultures. HIV-1 that was pre-incubated with DBM-2198 did not show any infectivity towards host cells whereas host cells pre-incubated with DBM-2198 remained susceptible to HIV-1 infection, suggesting that DBM-2198 acts on the virus particle rather than cell surface molecules in the inhibition of HIV-1 infection. Competition assays for binding to HIV-1 envelope protein with anti-gp120 and anti-V3 antibodies revealed that DBM-2198 acts on the viral attachment site of HIV-1 gp120, but not on the V3 region. This report provides a better understanding of the antiviral mechanism of DBM-2198 and may contribute to the development of a potential therapeutic drug against a broad spectrum of HIV-1 variants.

5-Hydroxytryptamine 6 Receptor (5-HT6R)-Mediated Morphological Changes via RhoA-Dependent Pathways

  • Rahman, Md. Ataur;Kim, Hanna;Lee, Kang Ho;Yun, Hyung-Mun;Hong, Jung-Hwa;Kim, Youngjae;Choo, Hyunah;Park, Mikyoung;Rhim, Hyewhon
    • Molecules and Cells
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    • v.40 no.7
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    • pp.495-502
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    • 2017
  • The $5-HT_6R$ has been considered as an attractive therapeutic target in the brain due to its exclusive expression in the brain. However, the mechanistic linkage between $5-HT_6Rs$ and brain functions remains poorly understood. Here, we examined the effects of $5-HT_6R$-mediated cell morphological changes using immunocytochemistry, Western blot, and live-cell imaging assays. Our results showed that the activation of $5-HT_6Rs$ caused morphological changes and increased cell surface area in HEK293 cells expressing $5-HT_6Rs$. Treatment with 5-HT specifically increased RhoA-GTP activity without affecting other Rho family proteins, such as Rac1 and Cdc42. Furthermore, live-cell imaging in hippocampal neurons revealed that activation of $5-HT_6Rs$ using a selective agonist, ST1936, increased the density and size of dendritic protrusions along with the activation of RhoA-GTP activity and that both effects were blocked by pretreatment with a selective $5-HT_6R$ antagonist, SB258585. Taken together, our results show that $5-HT_6R$ plays an important role in the regulation of cell morphology via a RhoA-dependent pathway in mammalian cell lines and primary neurons.

Adsorption and Photocatalytic Degradation of Dyes Using Synthesized Metal-Organic Framework NH2-MIL-101(Fe) (합성 금속-유기 골격체 NH2-MIL-101(Fe)를 이용한 염료의 흡착 및 광분해 제거)

  • Lee, Joon Yeob;Choi, Jeong-Hak
    • Journal of Environmental Science International
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    • v.27 no.7
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    • pp.611-620
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    • 2018
  • In this study, a metal-organic framework (MOF) material $NH_2$-MIL-101(Fe) was synthesized using the solvothermal method, and characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), UV-visible spectrophotometry, field-emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), and surface area measurements. The XRD pattern of the synthesized $NH_2$-MIL-101(Fe) was similar to the previously reported patterns of MIL-101 type materials, which indicated the successful synthesis of $NH_2$-MIL-101(Fe). The FT-IR spectrum showed the molecular structure and functional groups of the synthesized $NH_2$-MIL-101(Fe). The UV-visible absorbance spectrum indicated that the synthesized material could be activated as a photocatalyst under visible light irradiation. FE-SEM and TEM images showed the formation of hexagonal microspindle structures in the synthesized $NH_2$-MIL-101(Fe). Furthermore, the EDS spectrum indicated that the synthesized material consisted of Fe, N, O, and C elements. The synthesized $NH_2$-MIL-101(Fe) was then employed as an adsorbent and photocatalyst for the removal of Indigo carmine and Rhodamine B from aqueous solutions. The initial 30 min of adsorption for Indigo carmine and Rhodamine B without light irradiation achieved removal efficiencies of 83.6% and 70.7%, respectively. The removal efficiencies thereafter gradually increased with visible light irradiation for 180 min, and the overall removal efficiencies for Indigo carmine and Rhodamine B were 94.2% and 83.5%, respectively. These results indicate that the synthesized MOF material can be effectively applied as an adsorbent and photocatalyst for the removal of dyes.

A STUDY OF EFFECTS OF BONE MORPHOGENETIC PROTEIN BONE REGENERATION OF IMPLANTS IN DOGS (성견에서 임프란트 매식시 골형성단백 사용에 따른 골재생에 관한 연구)

  • Jo Jin-Hee;Vang Mong-Sook;Lee Jong-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.32 no.4
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    • pp.593-607
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    • 1994
  • The purpose of this study is to evaluate the effect of the bone morphogenetic protein, bone matrix gelatin and collagen matrix on the amount and shape of generating new bone adjacent to the implant. Implants were inserted in the mandible of adult dogs at 2 months after teeth extraction. Artificial bony defects, 3mm in width and 4mm in depth were made at the mesial and distal side of implant. Experimental groups were divided into three groups ; Group 1 : Defects filled with collagen matrix and bone morphogenetic protein, Group 2 : Defects filled with bone matrix gelatin. Control group : Defects filled with only collagen matrix. After implantation, the animals were sacrificed at 1,3,5 and 10 weeks for light microscopic examination. For the fluorescent microscopic examination. each tertracycline Hcl and calcein were injected at 1, 3, 5, 8 and 10 weeks after implantation. The results obtained were as follows : 1. The molecular weight of bovine BMP was about 18,100 by hydroxyapatite chromatography. 2. Osseointegration was observed in experimental groups 1 & 2, and BMG and BMP had an excellent bone forming capability as a filling materials to the repair of the bone defects. 3. The degree of healing of bone defect area, the experimental group 1 showed more prominent bone formation than control group, and the control group showed fibrous connective tissue between the implant and the bone. 4. In the fluorescent microscopic findings, bone remodelling was observed regenerative lamellar bone at defect area in experimental group 1, and partial remodelling in experimental group 2, In the control group, fibrous connective tissue was observed between the implant and bone surface and sign of remodelling was not apperaed. Above results suggest that BMP has rapid osteoinductive property and can be used clinically as a bone substitute on bone defects around implants.

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Cordycepin Suppresses Expression of Diabetes Regulating Genes by Inhibition of Lipopolysaccharide-induced Inflammation in Macrophages

  • Shin, Seul-Mee;Lee, Sung-Won;Kwon, Jeong-Hak;Moon, Sun-Hee;Lee, Seung-Jeong;Lee, Chong-Kil;Cho, Kyung-Hae;Ha, Nam-Joo;Kim, Kyung-Jae
    • IMMUNE NETWORK
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    • v.9 no.3
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    • pp.98-105
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    • 2009
  • Background: It has been recently noticed that type 2 diabetes (T2D), one of the most common metabolic diseases, causes a chronic low-grade inflammation and activation of the innate immune system that are closely involved in the pathogenesis of T2D. Cordyceps militaris, a traditional medicinal mushroom, produces a component compound, cordycepin (3'-deoxyadenosine). Cordycepin has been known to have many pharmacological activities including immunological stimulating, anti-cancer, and anti-infection activities. The molecular mechanisms of cordycepin in T2D are not clear. In the present study, we tested the role of cordycepin on the anti-diabetic effect and anti-inflammatory cascades in LPS-stimulated RAW 264.7 cells. Methods: We confirmed the levels of diabetes regulating genes mRNA and protein of cytokines through RT-PCR and western blot analysis and followed by FACS analysis for the surface molecules. Results: Cordycepin inhibited the production of NO and pro-inflammatory cytokines such as IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ in LPS-activated macrophages via suppressing protein expression of pro-inflammatory mediators. T2D regulating genes such as $11{\beta}$-HSD1 and PPAR${\gamma}$ were decreased as well as expression of co-stimulatory molecules such as ICAM-1 and B7-1/-2 were also decreased with the increment of its concentration. In accordance with suppressed pro-inflammatory cytokine production lead to inhibition of diabetic regulating genes in activated macrophages. Cordycepin suppressed NF-${\kappa}B$ activation in LPS-activated macrophages. Conclusion: Based on these observations, cordycepin suppressed T2D regulating genes through the inactivation of NF-${\kappa}B$ dependent inflammatory responses and suggesting that cordycepin will provide potential use as an immunomodulatory agent for treating immunological diseases.

ZnSe 박막 성장을 위한 Molecular Beam Epitaxy 성장 조건의 결정

  • Jeong, Myeong-Hun;Park, Seung-Hwan;Kim, Gwang-Hui;Jeong, Mi-Na;Yang, Min;An, Hyeong-Su;Jang, Ji-Ho;Kim, Hong-Seung;Song, Jun-Seok;Yao, Takafumi
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • v.9 no.1
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    • pp.990-994
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    • 2005
  • MBE growth conditions such as growth temperature, flux ration and growth rate for II-VI compound semiconductor growth has been studied. The growth temperature, flux ration and growth rate were tentatively controlled to 290$^{\circ}C$, 2, and 0.6 ${\mu}m$/h, respectively. From AFM result, relatively rough surface (RMS ${\sim}$ 2.9 nm) was observed. It was regarded as an indication of low growth temperature and high growth rate. XRD measurement shows that the film is relaxed, also the series of XRD measurements of different diffraction planes such as (002), (004), (115), (006) diffractions were performed to calculate the dislocation density in the film. The calculated dislocation density was found to be 8.30${\times}10^8$ dis/cm$^2$ which is compatible to the previous results.

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