• 제목/요약/키워드: ${\beta}$-Turn

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Inhibition of ClC-5 suppresses proliferation and induces apoptosis in cholangiocarcinoma cells through the Wnt/β-catenin signaling pathway

  • Shi, Zhe;Zhou, Liyuan;Zhou, Yan;Jia, Xiaoyan;Yu, Xiangjun;An, Xiaohong;Han, Yanzhen
    • BMB Reports
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    • v.55 no.6
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    • pp.299-304
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    • 2022
  • Chloride channel-5 (ClC-5), an important branch of the ClC family, is involved in the regulation of the proliferation and cell-fate of a variety of cells, including tumor cells. However, its function in cholangiocarcinoma (CCA) cells remains enigmatic. Here, we discovered that ClC-5 was up-regulated in CCA tissues and CCA cell lines, while ClC-5 silencing inhibited CCA cell proliferation and induced apoptosis. Further mechanism studies revealed that ClC-5 inhibition could inhibit Wnt/β-catenin signaling activity and further activate the mitochondria apoptotic pathway in CCA cells. Furthermore, rescuing Wnt/β-catenin signaling activation eliminated the anti-tumor function of ClC-5 knockdown. Together, our research findings illustrated that ClC-5 inhibition plays an anti-tumor role in CCA cells via inhibiting the activity of the Wnt/β-catenin pathway, which in turn activates the mitochondrial apoptotic pathway.

Dual-Toehold-Probe-Mediated Exonuclease-III-Assisted Signal Recycles Integrated with CHA for Detection of mecA Gene Using a Personal Glucose Meter in Skin and Soft Tissue Infection

  • Jiaguang Su;Wenjun Zheng
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1692-1697
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    • 2023
  • Staphylococcus aureus integrated with mecA gene, which codes for penicillin-binding protein 2a, is resistant to all penicillins and other beta-lactam antibiotics, resulting in poor treatment expectations in skin and soft tissue infections. The development of a simple, sensitive and portable biosensor for mecA gene analysis in S. aureus is urgently needed. Herein, we propose a dual-toehold-probe (sensing probe)-mediated exonuclease-III (Exo-III)-assisted signal recycling for portable detection of the mecA gene in S. aureus. When the target mecA gene is present, it hybridizes with the sensing probe, initiating Exo III-assisted dual signal recycles, which in turn release numerous "3" sequences. The released "3" sequences initiate catalytic hairpin amplification, resulting in the fixation of a sucrase-labeled H2 probe on the surface of magnetic beads (MBs). After magnet-based enrichment of an MB-H1-H2-sucrase complex and removal of a liquid supernatant containing free sucrase, the complex is then used to catalyze sucrose to glucose, which can be quantitatively detected by a personal glucose meter. With a limit of detection of 4.36 fM for mecA gene, the developed strategy exhibits high sensitivity. In addition, good selectivity and anti-interference capability were also attained with this method, making it promising for antibiotic tolerance analysis at the point-of-care.

Production of pediocin by Chemical Synthesis and Bactericidal Mode of Action

  • Koo, Min-Seon;Kim, Wang-June;Kwon, Dea-Young;Min, Kyung-Hee
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.149-153
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    • 2001
  • To investigate the mode of bactericidal action for antimicrobial peptide, pediocin, synthetic and mutant pediocins were prepared by direct chemical synthesis. Native pediocin was purified from Pedio-coccus acidilactici M and its conformational structure and bactericidal functions were analyzed and compared to synthetic pediocin. Schematic mode of pediocin actions, how pediocin binds on the target cell membrane, penetrates and makes tunnel are proposed. For these purposes, primary and secondary structures of pediocin was analyzed and disulfide bond assignment was also done. The pediocin purified from P. acidilactici M had high effective bactericidal ability against gram positive bacteria, especially Listeria monocytogenes and was very stable at extreme pHs and even at high temperatures such as autoclaving temperature (121$^{\circ}C$). Pediocin was consisted of 44 amino acids with four cysteines. Novel synthetic peptides were achieved by solid phase peptide synthesis(SPPS) method. To explain the function of cysteine in C-terminal region, mutant pediocin, Ped[C24A+C44A], was synthesized and their structural and biological functions were analyzed. Second mutant pediocin, Ped[KllE], was prepared to explain the function of lysine at 11 of N-terminal part of pediocin, especially loop of $\beta$-sheet, and to predict the initial binding site of pediocin. The native and synthetic pediocins was showed random coil conformation by spectropolarimetry in moderate conditions. This conformation was observed in extreme conditions such as high temperature and low and high pHs, also. Circular dichroism(CD) data also showed the existence of $\beta$-turn structure in N-terminal part both native and synthetic pediocins. A structural model for pediocin predicts that 18 amino acids in the N-terminal part of the peptide assume a three-strand $\beta$-sheet conformation. This random coil in C-terminal part of pediocin was converted to folding structure, helix structure, in nonpolar solvents such as alcohol and TFE. The disulfide bond between $^{9}$ Cys and $^{14}$ Cys was concrete and inevitable, however, evidences of disulfide bond between $^{24}$ Cys and $^{44}$ Cys was not. Data of Ped[C24A+C44A], pediocin mutant showed that $^{44}$ Cys was required during killing the target cells but not inevitable, since Ped[C24A+C44A] still have bactericidal activity but much less than native pediocin. Another pediocin mutant, Ped[KllE], had still bactericidal activity, was controversial to propose that positive charge like as $^{11}$ Lys in loop or hinge in bacteriocin bound or helped to binding to microorganism with electrostatic interaction between cell membrane especially teichoic acid and positive amino acid nonspecifically. The conformation of pediocin among native, synthetic and mutant pediocins did not show big difference. The conformations between oxidized and reduced pediocin were almost similar regardless of native or synthetic.

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Efficient Stereoselective Synthesis of (2S,3S,4S)-3,4-Dihydroxyglutamic Acid ((2S,3S,4S)-3,4-다이하이드록시글루타믹산의 효율적인 입체선택적 합성)

  • Jeon, Jongho;Shin, Nara;Lee, Jong Hyup;Kim, Young Gyu
    • Applied Chemistry for Engineering
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    • v.25 no.4
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    • pp.392-395
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    • 2014
  • (2S,3S,4S)-3,4-Dihydroxyglutamic acid (DHGA), a biologically active ${\alpha},{\beta}$-dihydroxy-${\gamma}$-amino acid, was efficiently synthesized from a readily available D-serine derivative in 30% overall yield over 11 steps. The key stereoselective $OsO_4$-catalyzed dihydroxylation reaction controlled by an N-diphenylmethylene group on the amino group of ${\gamma}$-amino-${\alpha},{\beta}$-unsaturated (Z)-ester successfully introduced the diol moiety of the intermediate 5a in 86% with more than 10 : 1 diastereomeric ration. Then it was in turn successfully converted to the desired target compound, (2S,3S,4S)-3,4-DHGA, via simple oxidation and hydrolysis in a highly stereoselective manner and a higher yield than the previous syntheses. This result strongly supports that our synthetic methodology of stereoselective $OsO_4$-catalyzed dihydroxylation should be useful in stereoselctive synthesis of various bioactive compounds with an amino diol moiety.

Relationship Between IFN${\gamma}$ Production, Antibody and Hormone Levels in Naturally Neospora caninum-infected Pregnant Dairy Cows (Neospora caninum에 자연 감염된 임신우에서 IFN${\gamma}$, 항체 및 호르몬 수준의 상관관계)

  • Son, Jeong-Hoon;Park, Bae-Keun;Son, Hwa-Young;Jung, Ju-Young;Park, Sang-Joon;Kim, Tae-Hwan;Cho, Sung-Whan;Ryu, Si-Yun
    • Journal of Veterinary Clinics
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    • v.27 no.6
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    • pp.640-646
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    • 2010
  • Neosporosis is a widespread parasitic disease caused by Neospora caninum, an intracellular protozoan parasite. It causes economic losses due to reproductive failure. The potential relationship between pregnancy outcomes and levels of IFN${\gamma}$, hormones, and antibodies in naturally N. caninum-infected cows was examined in the blood samples collected every 2 or 4 weeks in 26 pregnant cows from 4 different farms. The mean S/P value of seropositive nonaborting animals (n = 14) reached peak levels 15 weeks prior to parturition, and declined thereafter to parturition. The S/P value 13 weeks prior to abortion in seropositive aborting cows (n=3) remained at high levels, and abortions occurred at 20 (142 days), 26 (185 days), and 28 weeks (199 days) after artificial insemination. IFN${\gamma}$ levels in the seropositive non-aborting group varied by individuals and gestational periods; IFN${\gamma}$ levels stayed at elevated levels or increased abruptly close to abortion in seropositive aborting cows. IFN${\gamma}$ level patterns in the seronegative group (n = 9) were similar to the seropositive non-aborting group, although IFN${\gamma}$ amounts were lower than the seropositive group. The mean progesterone levels in the seropositive non-aborting and seronegative groups decreased markedly 7 weeks prior to parturition. The mean progesterone levels 5 and 7 weeks prior to abortion were lower than the other groups 5 and 7 weeks prior to parturition. The mean 17${\beta}$-estradiol levels in the seropositive aborting cows increased close to abortion; the produced amounts were lower than those of seropositive non-aborting and seronegative groups close to parturition. These results suggested that lower levels of progesterone and ${\beta}$-estradiol in Neospora-infected cows may lead to increases in IFN${\gamma}$ production and in turn may result in abortion.

The Effects of Metaphors in the Interface of Smartphone Applications on Users' Intention to Use (사용자환경의 메타포가 스마트폰 애플리케이션 사용의도에 미치는 영향)

  • Jung, Wonjin;Hong, Suk-Ki
    • Asia pacific journal of information systems
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    • v.24 no.3
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    • pp.255-279
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    • 2014
  • It is not too much to say that smartphones have become an essential part of our lives due to their versatility. Nevertheless, they still have less overall capabilities than their desktop counterparts. Specifically, they have small screens and low resolutions, which make their applications difficult to have a usable interface. To account for these limitations, the interface of smartphone applications should be designed carefully and properly. Good interface design to any application is critical. However, a comprehensive information systems (IS) literature review found that there has been little research on the user interface design of smartphone applications. More specifically, there has been little empirical evidence and understanding about how metaphors, an imaginative way of describing objects and concepts, in the user interface of smartphone applications affect users' intention to use the applications. Thereby, the research goals of this study are to examine 1) the effects of the metaphors in the user interface of smartphone applications on the interaction between users and applications and 2) the effects of mediating variables including the interaction between users and applications, users' beliefs and attitudes, on users' intention to use the applications. A survey was conducted to collect data. University students and practitioners participated in the survey. A 24-item questionnaire was developed on a 5-point Likert-type scale. The measurement items were mostly adapted from the previous studies in the IS literature and modified to fit the context of this study. First, a principal component factor analysis was performed to explore the inter-relationships among a set of variables. The analysis showed that most of the items loaded quite strongly on the six components. The analysis also revealed the six components with eigenvalues exceeding 1, explaining a total of 70.7 per cent of the variance. The reliabilities of the items were also checked. Most Cronbach alpha values were above 0.8, so the scales were considered reliable. In sum, the results of the analysis support the decision to retain the six factors for further investigation. Next, the structural model was analyzed with AMOS structural equation modeling. The values of GFI, AGFI, NFI, TLI, CFI, and RMSEA were checked. The values showed that the research model considerably have a good fit in general. Next, the convergent and discriminant validities of all constructs were examined. The values for the standardized regression weights and critical ration (CR) indicated sufficient convergent validity for all constructs. In addition, the square root of the average variance extracted (AVE) of each construct was compared with its correlations with all other constructs. The results supported discriminant validity for all constructs. In sum, the results of analysis demonstrated adequate convergent and discriminant validities for all constructs. Finally, path coefficients between the variables were examined. Methphor was found to have an impact on interaction (${\beta}$ = .457, p = .000). There were also significant effects of the interaction on perceived usefulness (${\beta}$ = .273, p = .000) and ease of use (${\beta}$ = .405, p = .000). User attitude was significantly influenced by these two beliefs, perceived usefulness (${\beta}$ = .386, p = .000) and ease of use (${\beta}$ = .347, p = .000) respectively. Further, the results of analysis found that users' intention to use smartphone applications was significantly influenced by user attitude (${\beta}$ = .567, p = .000). Based upon the analyses, all hypotheses were supported. This study found that the metaphors used in the interface of smartphone applications affect not only the interaction between users and applications, but also users' intention to use the applications through the mediating variables, perceived usefulness and ease of use. These findings imply that if the metaphors used in the user interface of application are easy enough to understand for smartphone users, then the application can be perceived useful and easy to use, which in turn make users to have an intention to use the application. In conclusion, this study contributed not only to validate and extend Technology Acceptance Model (TAM) partially, but also to develop the construct of metaphor in smartphone settings. However, since a single empirical study cannot be enough to validate the findings, some limitations should be considered.

The Evaluation of Reliability for Exam Distance of Visual Acuity (시력검사거리에 따른 원거리 시력검사 신뢰성 평가)

  • Chun, Young-Yun;Choi, Hyun-Soo;Park, Seong-Jong;Lee, Seok-Ju
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.1
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    • pp.17-22
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    • 2014
  • Purpose: We aimed to evaluate reliability of eye exam for visual acuity as a function of distance. Methods: There were 39 patients (78 eyes) who had visual acuity 1.0 or more at 5 meters. We measured refractive power of patients at each distances, 5 meters, 4 meters and 3 meters. Automatic chart (LCD-700, Hyeseong Optic. Co., Korea) used for visual acuity, skiascope (Beta 200, Heine, Germany) and auto refractometer (RK-5, Canon, Japan) used as for objective refraction. Accommodation was examined by minus lens addition methods, and Accommodative lag was examined by grid chart for reading distance. Results: Being compared to 3 meter test, Amount of corrected spherical refractive power decreased by $0.10{\pm}0.38$ D, astigmatism decreased by $0.05{\pm}0.10$ D, and axis of astigmatism rotated toward to temporal by $2.64{\pm}18.75$ degrees for right eyes, by $11.43{\pm}48.55$ degrees for left eyes in case of 5 meter test. Changes of corrected refraction and astigmatism were slightly correlated (r=-0.31, r=-0.29). Conclusions: Because corrected refraction power and amount of astigmatism decreased and axis of astigmatism tends to turn the temporal direction according to exam distance, examination distance of visual acuity should improved as to 5 meters.

Effect of Gamma Irradiation on the Structural and Physiological Properties of Silk Fibroin

  • Sung, Nak-Yun;Byun, Eui-Baek;Kwon, Sun-Kyu;Kim, Jae-Hun;Song, Beom-Seok;Choi, Jong-Il;Kim, Jin-Kyu;Yoon, Yo-Han;Byun, Myung-Woo;Kim, Mee-Ree;Yoo, Hong-Sun;Lee, Ju-Woon
    • Food Science and Biotechnology
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    • v.18 no.1
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    • pp.228-233
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    • 2009
  • This study was conducted to examine the changes in the molecular structure and physiological activities of silk fibroin by gamma irradiation. The results of gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the molecular weight of fibroin was increased depending upon the irradiation dose. Secondary structure of fibroin determined by using circular dichroism revealed that the ratio of $\alpha$-helix was increased up to 10 kGy and then decreased depending upon the irradiation dose. Whereas, the ratio of $\beta$-sheet, $\beta$-turn, and random coil were decreased and then increased with an alteration in the $\alpha$-helix secondary conformation. The 2.2-diphenyl-1-picryl-hydrazil (DPPH) radical scavenging activity of fibroin was increased by gamma irradiation at 5 kGy, but was decreased above 10 kGy depending upon the irradiation dose. Also, the inhibition activities of tyrosinase and melanin synthesis of fibroin were increased by gamma irradiation. These results indicated that gamma irradiation could be used as an efficient method to make fibroin more suitable for the development of functional foods and cosmetics.

Agastache rugosa Leaf Extract Inhibits the iNOS Expression in ROS 17/2.8 Cells Activated with TNF-$\alpha$ and IL-$\beta$

  • Oh Hwa Min;Kang Young Jin;Kim Sun Hee;Lee Young Soo;Park Min Kyu;Heo Ja Myung;Sun Jin Ji;Kim Hyo Jung;Kang Eun Sil;Kim Hye Jung;Sea Han Geuk;Lee Jae Heun;YunChoi Hye Sook
    • Archives of Pharmacal Research
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    • v.28 no.3
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    • pp.305-310
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    • 2005
  • It has been suggested that nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) may act as a mediator of cytokine-induced effects on bone turn-over. NO is also recognized as an important factor in bone remodeling, i.e., participating in osteoblast apoptosis in an arthritic joint. The components of Agastache rugosa are known to have many pharmacological activities. In the present study, we investigated the effects of Agastache rugosa leaf extract (ELAR) on NO production and the iNOS expression in ROS 17/2.8 cells activated by a mixture of inflammatory cytokines including TNF-$alpha$ and IL-1$\beta$. A preincubation with ELAR significantly and concentration-dependently reduced the expression of iNOS protein in ROS 17/2.8 cells activated with the cytokine mixture. Consequently, the NO production was also significantly reduced by ELAR with an IC$_{50}$ of 0.75 mg/mL. The inhibitory mechanism of iNOS induction by ELAR prevented the activation and translocation of NF-$\kappa$B (p65) to the nucleus from the cytosol fraction. Furthermore, ELAR concentration-dependently reduced the cellular toxicity induced by sodium nitroprusside, an NO-donor. These results suggest that ELAR may be beneficial in NO-mediated inflammatory conditions such as osteoporosis.

Changes of Organic Acids and Volatile Flavor Compounds of Traditional Andong Sikhe (전통안동식혜의 유기산 및 휘발성 향기성분의 변화)

  • Choi, Cheong;Woo, Hi-Seob;An, Bong-Jeun;Cho, Young-Je;Kim, Seung
    • Journal of the Korean Society of Food Culture
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    • v.10 no.1
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    • pp.11-17
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    • 1995
  • The changes in life style these days appear in many ways. Many housewives turn away from home preparation of the time consuming traditional foods, such as Andong Sikhe. The importance, however, of succeeding the traditional cuisines is getting appreciated widely now a days. A traditional Andong Sikhe in Korea, was prepared and fermented at $5^{\circ}C$ in order to analyze the taste and flavor compounds. Major flavor components were identified as camphene, sabinene, 1-(1,5-dimethyl-4-hexyl)-4-methyl-benzene, ${\alpha}-zingibirene$, farnesene, 2,6-bis(1,1-dimethylethyl)-4-methyl-phenol, ${\beta}-sesquiphellandrene$, calalene, tetradecanoic acid, hexadecanoic acid and 9,12-octadecanoic acid by GC and GC/MS. Lactic acid and increased from 0.49 mg/100 g in the initial stage to 19.37 mg/100 g at the 6 days of storage. The pH of product decresed to 3.7 from 5.4 after the 6 days of storage but the total acidty reached to 0.41% during fermentation and storage at $5^{\circ}C$.

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