• Title/Summary/Keyword: VPS34

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Bacterial Overexpression and Denaturing Purification of VPS34-Binding Domain of Beclin 1

  • Baek, Jong-Hyuk;Jung, Juneyoung;Seo, Jeongbin;Kim, Jeong Hee;Kim, Joungmok
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1808-1816
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    • 2016
  • As a scaffolding subunit of the PIK3C3/VPS34 complex, Beclin 1 recruits a variety of proteins to class III phosphatidylinositol-3-kinase (VPS34), resulting in the formation of a distinct PIK3C3/VPS34 complex with a specific function. Therefore, the investigation of a number of Beclin 1 domains required for the protein-protein interactions will provide important clues to understand the PIK3C3/VPS34 complex, of which Beclin1-VPS34 interaction is the core unit. In the present study, we have designed a bacterial overexpression system for the Beclin 1 domain corresponding to VPS34 binding (Vps34-BD) and set up the denaturing purification protocol due to the massive aggregation of Vps34-BD in Escherichia coli. The expression and purification conditions determined in this study successfully provided soluble and functional Vps34-BD.

A Study on Applying Genetic Algorithm for Optimum Virtual Path Network Design in ATM Network (ATM 망에서 최적 가상 경로망 설계를 위한 유전자 알고리즘 응용에 관한 연구)

  • 강주락;권기호
    • Proceedings of the IEEK Conference
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    • 2000.06c
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    • pp.31-34
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    • 2000
  • Genetic Algorithm is well known as the efficient algorithm which can solve a difficult optimization problem. Recently, there has been increasing interest in applying genetic algorithm to problem related to network design. In this paper, we propose a two-step genetic algorithm for designing a optimum virtual path network(VPN) for a given physical network and traffic demand. The first step is a routing step in which a route is found between every node pair in the network. In the second step, paths are assigned as VPs so as to minimize the total number of VPs configured, the number of VPs carried by a link, and the VP hopcount. We study the performance of the propose algorithm through simulation. The result shows that the VPN generated by the proposed algorithm is good in minimizing the number of VPs configured, the load on a link, and the VP hopcount.

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Study on the Surface Properties of Corneocyte between Face and Forearm Using Atomic Force microscopy (AFM) (AFM을 이용한 얼굴과 하박내측 각질세포 표면 특성 비교연구)

  • Chang, Minyoul
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.4
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    • pp.373-380
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    • 2019
  • There are many differences in tran-epidermal water loss (TEWL), skin water contents, and skin elasticity, etc between face and forearm skin. In particular, our previous studies showed that elasticity of face skin was significantly differed from forearm depending on full hydration. So, we have studied the surface properties of corneocyte using atomic force microscopy (AFM), assuming that the differences between face and forearm skin would be associated with the surface properties of corneocyte. The surface roughness of corneocyte and villus-like projections (VPs) were measured. Furthermore, qualitative comparison among the surface of face, forearm, and lip corneocyte was performed. Corneocytes were collected by tape-stripping on both face and forearm of 8 volunteers, and the bottom surface of corneocytes were measured at 40 ㎛ × 40 ㎛ using AFM. Results showed that the lower surface roughness of face corneocytes was 388.34 ± 86.189 nm, and that of forearm corneocytes was 662.27 ± 224.257 nm, which confirmed that the lower surface of forearm corneocytes was more rough than that of face corneocytes (p < 0.001). Compared with the amount of VPs, lip corneocytes were the highest followed by face corneocytes, and forearm corneocytes were the lowest. From these results, it is conclued that the surface properties of corneocytes are somewhat involved in the property differences between the face and the forearm skin and VPs can be a useful parameter for the study of corneocyte by site. In addition, AFM is a very useful device for the comparative study of nano-structural differences on the surface of corneocytes. More studies can lead to develop a new evaluation method of corneocytes.

Hepatitis B virus X Protein Promotes Liver Cancer Progression through Autophagy Induction in Response to TLR4 Stimulation

  • Juhee Son;Mi-Jeong Kim;Ji Su Lee;Ji Young Kim;Eunyoung Chun;Ki-Young Lee
    • IMMUNE NETWORK
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    • v.21 no.5
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    • pp.37.1-37.17
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    • 2021
  • Hepatitis B virus X (HBx) protein has been reported as a key protein regulating the pathogenesis of HBV-induced hepatocellular carcinoma (HCC). Recent evidence has shown that HBx is implicated in the activation of autophagy in hepatic cells. Nevertheless, the precise molecular and cellular mechanism by which HBx induces autophagy is still controversial. Herein, we investigated the molecular and cellular mechanism by which HBx is involved in the TRAF6-BECN1-Bcl-2 signaling for the regulation of autophagy in response to TLR4 stimulation, therefore influencing the HCC progression. HBx interacts with BECN1 (Beclin 1) and inhibits the association of the BECN1-Bcl-2 complex, which is known to prevent the assembly of the pre-autophagosomal structure. Furthermore, HBx enhances the interaction between VPS34 and TRAF6-BECN1 complex, increases the ubiquitination of BECN1, and subsequently enhances autophagy induction in response to LPS stimulation. To verify the functional role of HBx in liver cancer progression, we utilized different HCC cell lines, HepG2, SK-Hep-1, and SNU-761. HBx-expressing HepG2 cells exhibited enhanced cell migration, invasion, and cell mobility in response to LPS stimulation compared to those of control HepG2 cells. These results were consistently observed in HBx-expressed SK-Hep-1 and HBx-expressed SNU-761 cells. Taken together, our findings suggest that HBx positively regulates the induction of autophagy through the inhibition of the BECN1-Bcl-2 complex and enhancement of the TRAF6-BECN1-VPS34 complex, leading to enhance liver cancer migration and invasion.

COMPARISON OF STONE SURFACE POROSITIES CAUSED BY HYDROGEN GAS FROM ADDITION SILICONE IMPRESSION MATERIALS (부가 중합형 실리콘 인상재에서 발생하는 수고 기체가 경석고 표면에 미치는 영향)

  • Yoo, So-Jeong;Lee, Keun-Woo;Kim, Kyeung-Nam
    • The Journal of Korean Academy of Prosthodontics
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    • v.34 no.2
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    • pp.349-362
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    • 1996
  • To determine the factors to affect on stone surface porosities produced from hydrogen gas of additional silicone, both putty and syringe type of 7 commercially different additional silicone impression materials(Blend-A-Scon, Correct VPS, Exaflex, Express, Extrude, Provil, Reprosil) were chosen and NewFujirock(GC) was poured into the impressions of detail-reproducing test block at 1, 15, 30, 45, 60 minutes after the impression materials had set and 4 specimens were made for each pouring time, each type of impression material, and each consisency and So, 280 specimens were made in total. The number of surface porosities of same area($2826 mm^2$) which were typically caused by hydrogen gas using the stereoscope(X 7.5) by two observers. Comparison of putty-syringe type and among the impression materials are tested by Kruscal-Wallis method and Mann-Whitney method(p<0.05). The results are as follows. 1. The number of porosities decreased as the pouring time of stone was delayed on both putty and syringe type of additional silicone materials. 2. The putty type significantly produced more porosities than syringe type except for the group of Reprosil.(p<0.05). 3. In case of putty type, the number of porosities increased as following order. Reprosil / Blend-A-Scon and Provil / Correct VPS and Extrude / Express and Exaflex. 4. In case of syringe type, Blend-A-Scon and Extrude produced no porosity and Exaflex and Provil at 30 minites, but Express produced porosities even at 60 minutes and the most. Additional silicone impression material releases hydrogen gas, and that fact can make the resulting die stone model useless. So, to minimize these adverse effects, it is desirable not to expose putty type of additional silicone on critical impression surface because putty type has a tendency to produce more porosities than syringe type. And it is important to have sufficient time before pouring the stone on impression because porosities produce less as time passes after setting of impression material. Also, there are differences among 7 additional silicone impression materials, so it is desirable to choose adequate brand of additional silicone for good laboratory work.

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HMGB1 regulates autophagy through increasing transcriptional activities of JNK and ERK in human myeloid leukemia cells

  • Zhao, Mingyi;Yang, Minghua;Yang, Liangchun;Yu, Yan;Xie, Min;Zhu, Shan;Kang, Rui;Tang, Daolin;Jiang, Zhigang;Yuan, Wuzhou;Wu, Xiushan;Cao, Lizhi
    • BMB Reports
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    • v.44 no.9
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    • pp.601-606
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    • 2011
  • HMGB1 is associated with human cancers and is an activator of autophagy which mediates chemotherapy resistance. We here show that the mRNA levels of HMGB1 are high in leukemia cells and it is involved in the progression of childhood chronic myeloid leukemia (CML). HMGB1 decreases the sensitivity of human myeloid leukemia cells K562 to anti-cancer drug induced death through up-regulating the autophagy pathway, which is confirmed by the observation with an increase in fusion of autophagosomes and autophagolysosomes. When overexpressing HMGB1, both mRNA levels of Beclin-1, VSP34 and UVRAG which are key genes involved in mammalian autophagy and protein levels of p-Bcl-2 and LC3-II are increased. Luciferase assays document that over-expression of HMGB1 increases the transcriptional activity of JNK and ERK, which may be silenced by siRNA. The results suggest that HMGB1 regulates JNK and ERK required for autophagy, which provides a potential drug target for therapeutic interventions in childhood CML.

Rutin induces autophagy in cancer cells

  • Park, Mi Hee;Kim, Seyeon;Song, Yu-ri;Kim, Sumi;Kim, Hyung-Joon;Na, Hee Sam;Chung, Jin
    • International Journal of Oral Biology
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    • v.41 no.1
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    • pp.45-51
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    • 2016
  • Rutin (3,3',4',5,7-pentahydroxyflavone-3-rhamnoglucoside) is a bioactive flavonoid from the plant kingdom. Rutin has been studied as potential anticancer agent due to its wide range of pharmacological properties including antioxidative, anti-inflammatory and anticancer. Autophagy is a conserved intracellular catabolic pathway to maintain cell homeostasis by formation of autophagosome. Processing of autophagy involves various molecules including ULK1 protein kinase complex, Beclin-1-Vps34 lipid kinase complex, ATG5, ATG12, and LC3 (light chain 3). Cargo-carried autophagosomes fuse with lysosomes resulting in autophagolysosome to eliminate vesicles and degrade cargo. However, the actions of rutin on autophagy are not clearly understood. In this study, we analyzed the effect of rutin on autophagy and inflammation in cancer cell lines. Interestingly, rutin induced autophagy in leukemia (THP-1), oral (CA9-22), and lung (A549) cell lines. TNF-${\alpha}$, key modulator of inflammation, was upregulated by inhibition of rutin-induced autophagy. Taken together, these data indicated that rutin induced autophagy and consequently suppressed TNF-${\alpha}$ production.

EFFECT OF CONTAMINANTS ON THE PUTTY-WASH BOND STRENGTH IN TWO-STEP RELINE TECHNIQUE USING VINYL POLYSILOXANE IMPRESSION MATERIALS (Vinyl Polysiloxane 인상재를 이용한 이회 인상법에서 contaminants가 putty-wash 결합력에 미치는 영향)

  • Kim, Mu-Hyon;Jeong, Chang-Mo;Jeon, Young-Chan;Hwang, Hie-Seong
    • The Journal of Korean Academy of Prosthodontics
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    • v.34 no.2
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    • pp.266-276
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    • 1996
  • Numerous factors are known to affect the accuracy of elastomeric impression materials. Factor often overlooked is the quality of the bond between putty and wash during corrective reline impression technique. The putty-wash bond strength must be strong enough to over-come the local stress at putty-wash interface. It is not always possible to avoid saliva contamination in making corrective wash impres-sion. And putty preliminary impression material con be used as a template for provisional restoration. Human saliva and the residual monomer of autopolymerizing acrylic resin are thought to affect the bond strength and the failure type. This study examined the effect of contaminants like human saliva, and residual resin monomer on the putty-wash bond strength and the effectiveness of treatment. 1. Of the tested three brands of Vinyl Polysiloxane impession meterial, Express Exhibited the greatest bond strength followed by Eamix and Perfect showed the lowest putty-wah bond strength. 2. Coating the putty substrates with human saliva did not produce decreased failure load in all the breands of Vinyl Polysiloxane impression meterail. 3. Of the three brands of VPS impression material that were exposed to methhylmethacry-late resin(Jet), only the putty-wash bond strength of the Perfect group diminished signifi-cantly. Moreover, all the specimens from group C of Perfect exhibited adhesive failure. 4. Exposing the substrates to ethylmethacrylate resin(SNAP. diminished the putty-wash bond strength significantly. With Perfect and Examix, failure occurred cohesively through the light-body, whereas with Express, failure occurred adhesive-cohesively. 5. Removing approximately 1mm thickness of the contaminated putty interface was the most effective treatment in countering the undesirable effect caused by residual resin monomer. The putty-wash bond strength of the groups that were treated with 1mm even putty reduction was not significantly different from those of control groups. With Perfect and Examix, cleaning the specimens with gauze soaked in 70% isopropyl alcohol increased the putty-wash bond strength, but was not as effective as 1mm even reduction of contaminated putty substrates. With Express, 70% isoproryl alcohol treatment exhibi0ted comparable putty-wash bond strength to that of control group.

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