• Title/Summary/Keyword: Nanosensor

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Expression and Purification of Recombinant Active Prostate-Specific Antigen from Escherichia coli

  • Jeong, Su-Jin;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.840-846
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    • 2007
  • Human prostate-specific antigen(PSA), a 33 kDa serine protease with comprehensive homology to glandular kallikrein, is secreted from prostatic tissue into the seminal fluid and enters into the circulation. The level of PSA increases in the serum of patients with prostatic cancer and hence is widely employed as a marker of the disease status. In particular, an enzymatically active PSA that is a form cleaved at the N-terminal seven-amino-acids prosequence, APLILSR, of proPSA may play an important roll in the progression of prostate cancer. Thus, the presence of the active form would selectively discriminate the cancer from benign prostatic hyperplasia. In this study, we developed a convenient purification method for the acquisition of active PSA and proPSA. Recombinant proPSA and active PSA were expressed directly in Escherichia coli, easily and efficiently isolated from inclusion bodies, refolded, and purified. Moreover, the enzymatic activity of the recombinant active PSA was confirmed as serine protease using chromogenic chymotrypsin substrate. This purified active PSA could be further applied to scrutinize the biological or conformational characteristics of the protein and to develop specific diagnostic and/or therapeutic agents against prostate cancer.

Vibration analysis thermally affected viscoelastic nanosensors subjected to linear varying loads

  • Ebrahimi, Farzad;Babaei, Ramin;Shaghaghi, Gholam Reza
    • Advances in nano research
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    • v.6 no.4
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    • pp.399-422
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    • 2018
  • Unwanted vibration is an issue in many industrial systems, especially in nano-devices. There are many ways to compensate these unwanted vibrations based on the results of the past researches. Elastic medium and smart material etc. are effective methods to restrain unnecessary vibration. In this manuscript, dynamic analysis of viscoelastic nanosensor which is made of functionally graded (FGM) nanobeams is investigated. It is assumed that, the shaft is flexible. The system is modeled based on Timoshenko beam theory and also environmental condition, external linear varying loads and thermal loading effect are considered. The equations of motion are extracted by using energy method and Hamilton principle to describe the translational and shear deformation's behavior of the system. Governing equations of motion are extracted by supplementing Eringen's nonlocal theory. Finally vibration behavior of system especially the frequency of system is developed by implementation Semi-analytical differential transformed method (DTM). The results are validated in the researches that have been done in the past and shows good agreement with them.

Functional Analysis of a Grapevine UDP-Glucose Flavonoid Glucosyl Transferase (UFGT) Gene in Transgenic Tobacco Plants (담배 형질전환체를 이용한 포도 UDP-glucose flavonoid glucosyl transferase (UFGT) 유전자의 기능 분석)

  • Park, Ji-Yeon;Park, Sung-Chool;Pyee, Jae-Ho
    • Journal of Life Science
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    • v.20 no.2
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    • pp.292-297
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    • 2010
  • Anthocyanin, a phenolic compound, is a pigment that shows blue or red color in the fruit, petal and other tissues. It is an important factor in grape berry skin pigment and accumulates only in the skin. This skin-specific accumulation of anthocyanin has been reported to be regulated by the ufgt gene which encodes UDP-glucose: flavonoid 3-O-glucosyltransferase that participates in the biosynthesis of anthocyanin. The ufgt gene is expressed only in berry skin, while the other genes involved in the biosynthetic pathway are expressed in both skin and flesh tissues. In order to determine whether anthocyanin accumulation is primarily regulated by compartment of UFGT, a ufgt cDNA clone was isolated from grape berry, its open reading frame was ligated in pBI121 vector in either a sense or an antisense orientation under the control of the CaMV35S promoter and the recombinant constructs were incorporated into tobacco plants. Several transgenic lines were selected and characterized to determine the level of expression of the grapevine ufgt transcript and endogenous homologs of tobacco. Compared to the wild-type, the amount of anthocyanins in sense transgenic plants increased by 44%, while the amount of anthocyanins in antisense transgenic plants decreased by 88%. In addition, the color of flowers became intense in the sense transgenic plants. These results suggest that over-expression or repression of the ufgt gene affected the accumulation of anthocyanin in flowers of tobacco.

Effects of Ethylene Precursor, Auxin and Methyl Jasmonate on the Aerenchyma Formation in the Primary Root of Maize (Zea mays) (옥수수(Zea mays) 원뿌리의 통기조직 발달에 미치는 에틸렌 전구체, 옥신, 메틸자스몬산의 효과)

  • Ho, Jongyoon;Maeng, Sohyun;Park, Woong June
    • Journal of Life Science
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    • v.25 no.1
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    • pp.37-43
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    • 2015
  • We have investigated the effects of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), indole-3-acetic acid (IAA) and methyl jasmonate (MeJA) on the development of aerenchyma in the primary root of maize (Zea mays). Because plant hormones affected the longitudinal organization of the primary root, we need an indicator to direct the positions for comparison between control and hormone-treated roots. Therefore, the zones of the maize primary root were categorized as PR25, PR50 and PR75, where each value indicates the relative position between the root tip (PR0) and the base (PR100). Aerenchyma was not observed at PR25 and PR50 and rarely found at PR75 in the cortex of control roots. The aerenchymal area at PR75 increased in the presence of the ethylene precursor ACC or a natural auxin IAA. On the other hand, MeJA differentially acted on non-submerged and submerged roots. Exogenously applied MeJA suppressed the aerenchyma formation in non-submerged roots. When the primary root was submerged, aerenchymal area expanded prominently. The submergence-induced aerenchyma formation was amplified with MeJA. Lateral root primordia have been known to inhibit aerenchymal death of surrounding cells. All the three hormones stimulating aerenchyma formation as described above did not restore the inhibition caused by lateral root primordia, suggesting that the inhibitory step regulated by lateral root primordia can be located after hormonal signaling steps.

Effect of Fermented Garlic Extract on Adhesion of THP-1 Cells to Bovine Aortic Endothelial Cells (혈관내피세포에 부착하는 THP-1에 대한 발효마늘추출물의 효과)

  • Kwon, Ohseong;Song, Jina;Yu, Heui Jong;Park, Heonyong
    • Journal of Life Science
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    • v.27 no.5
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    • pp.553-560
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    • 2017
  • Fermented garlic extract (FGE) is a well-known functional food containing cycloalliin. Here we tested if FGE has an effect on the vascular system. We found that FGE had an effect on monocytic adhesion to endothelial cells, which occurs at the initial step of processes of inflammation. From dose-dependent experiments, 0.1 to $1{\mu}g/ml$ of FGE appeared to inhibit lipopolysaccharide (LPS)-enhanced heterotypic cell adhesion between THP-1 and BAECs. This result indicates that FGE blocks vascular inflammation. Then we found that FGE activates eNOS and Akt in BAECs. The phosphorylation of eNOS was maximally elevated 10 min after FGE treatment. Parallely, the phosphorylation of Akt was also maximally increased 10 min after FGE treatment. Consistently, it was found that FGE enhanced the production of nitric oxide. We then examined whether NO mediates THP-1 cell adhesion to BAECs. Both Akt and eNOS inhibitors appeared to reverse an inhibitory effect of FGE. These findings indicate that FGE inhibits LPS-enhanced heterotypic cell adhesion via Akt and eNOS. In conclusion, FGE plays an important role in prevention of inflammatory diseases.

The Effect of Sodium Tungstate on the Aldehyde Oxidase and the Growth in the Primary Root of Maize (Zea mays) (옥수수 (Zea mays) 뿌리의 알데히드 산화효소와 생장에 미치는 텅스텐산 나트륨의 영향)

  • Oh, Young-Joo;Cho, Young-Jun;Park, Woong-June
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.990-995
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    • 2007
  • We tested the effect of sodium tungstate, which disturbs the molybdenum cofactor formation, on the activities of aldehyde oxidase(AO) and the growth of maize(Zea mays) primary roots. As reported in other plants, sodium tungstate inhibited AO also in the maize root concentration-dependently. The inhibitory effect of sodium tungstate was observed only when the inhibitor was applied to the living plants. Application of tungstate to the extracted protein did not show any effect. Western analysis revealed slightly decreased level of AO protein in the presence of tungstate, indicating a positive feedback of gene regulation by the product. We also tested the effects of tungstate on the root growth. The elongation of primary root and the development of lateral roots, which are sensitive to the absolute level of auxin, were decreased in the presence of sodium tungstate. However, the gravitropic curvature of the primary root, which is dependent on the relative amount of auxin at both sides, was unaffected. These data suggested the decrease of auxin biosynthesis by the application of tungstate. However, the level of free IAA was unaffected by tungstate application. We discuss the possible explanations for the observed results.

Supply of Tryptophan and Tryptamine Influenced the Formation of Melatonin in Viola Plants (제비꽃속(Viola) 식물에서 tryptophan과 tryptamine 공급이 멜라토닌 생성에 미치는 영향)

  • Kim, Yeo-Jae;Yoon, Young-Ha;Park, Woong-June
    • Journal of Life Science
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    • v.21 no.2
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    • pp.328-333
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    • 2011
  • Melatonin has been known as an animal hormone. However, melatonin exists in diverse organisms including higher plants. The biosynthesis and physiological roles for melatonin in plants is still largely unknown, although both dicot and monocot plants have melatonin and some medicinal plants even contain large amounts of melatonin. In this study we detected melatonin in diverse Viola plants, in which melatonin had not been examined so far, by reverse phase HPLC analysis, demonstrating the wide existence of melatonin in the genus of Viola. We then fed tryptophan (Trp) and tryptamine (TAM) to the incubation medium for Viola leaf sections to test their effects on melatonin formation. Trp is also the hypothesized starting material of melatonin in plants, and TAM is the following intermediate produced by the decarboxylation of Trp. Trp feeding did not affect the contents of melatonin. In contrast, TAM feeding clearly increased the level of melatonin in Viola leaves. Because TAM is derived from Trp, we concluded that the Trp-TAM pathway exists in Viola plants as well. Ineffectiveness of Trp feeding to the change of melatonin contents supports the hypothesis that the decarboxylation step from Trp to TAM is the rate-limiting step in plant melatonin biosynthesis.

Characterization of the ZnSe/ZnS Core Shell Quantum Dots Synthesized at Various Temperature Conditions and the Water Soluble ZnSe/ZnS Quantum Dot

  • Hwang, Cheong-Soo;Cho, Ill-Hee
    • Bulletin of the Korean Chemical Society
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    • v.26 no.11
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    • pp.1776-1782
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    • 2005
  • ZnSe/ZnS, UV-blue luminescent core shell quantum dots, were synthesized via a thermal decomposition reaction of organometallic zinc and solvent coordinated Selenium (TOPSe) in a hot solvent mixture. The synthetic conditions of the core (ZnSe) and the shell (ZnS) were independently studied at various reaction temperature conditions. The obtained colloidal nanocrystals at corresponding temperatures were characterized for their optical properties by UV-vis, room temperature solution photoluminescence (PL) spectroscopy, and further obtained powders were characterized by XRD, TEM, and EDXS analyses. The synthetic temperature condition to obtain the best PL emission intensity for the ZnSe core was 300 ${^{\circ}C}$, and for the optimum shell capping, the temperature was 135 ${^{\circ}C}$. At this temperature, solution PL spectrum showed a narrow emission peak at 427 nm with a PL efficiency of 15%. In addition, the measured particle sizes for the ZnSe/ZnS nanocomposite via TEM were in the range of 5 to 12 nm. Furthermore, we have synthesized water-soluble ZnSe/ZnS nanoparticles by capping the ZnSe/ZnS hydrophobic surface with mercaptoacetate (MAA) molecules. For the obtained aqueous colloidal solution, the UV-vis spectrum showed an absorption peak at 250 nm, and the solution PL emission spectrum showed a peak at 425 nm, which is similar to that for hydrophobic quantum dot ZnSe/ZnS. However, the calculated PL efficiency was relatively low (0.1%) due to the luminescence quenching by water and MAA molecules. The capping ligand was also characterized by FT-IR spectroscopy, with the carbonyl stretching peak in the mercaptoacetate molecule appearing at 1575 $cm ^{-1}$. Finally, the particle sizes of the MAA capped ZnSe/ZnS were measured by TEM, showing a range of 12 to 17 nm.

Glass Transition Behavior of Dendritic Polymers Containing Mobile Aliphatic Polyether Cores and Glassy Peripheral Polystyrenes

  • Song, Jie;Cho, Byoung-Ki
    • Bulletin of the Korean Chemical Society
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    • v.29 no.6
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    • pp.1167-1172
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    • 2008
  • We investigated the glass transition temperatures ($T_g$) of dendrons consisting of conformationally mobile aliphatic polyether dendritic cores plus glassy peripheral polystyrenes (PSs), and linear PSs in the molecular weight range of 1000-8500 g/mol. We compared their $T_g$ behavior depending on their polymeric architecture. The linear PSs show a typical growth of $T_g$ up to 92.5 ${^{\circ}C}$ as the molecular weight increases to 8300 g/mol, while the dendrons display nearly constant $T_g$ values of 58-61 ${^{\circ}C}$, despite the increase of molecular weight with each generation. The striking contrast of Tg behavior would be mainly attributed to the fact that the dendrons keep the ratio of $N_e$/M ($N_e$: number of peripheral chain ends, M: molecular weight) over all the generations. Additionally, for the influence of dendritic spacers on glass transition temperature we prepared dimeric PSs with different linkage groups such as aliphatic ether, ester and amide bonds. We found that the dimer with the ether spacer exhibited the lowest glass transition at 55.4 ${^{\circ}C}$, while the amide linked dimer showed the highest glass transition temperature at 74.2 ${^{\circ}C}$. This indicates that the peripheral PS chains are effectively decoupled by the conformationally flexible ether spacer. The results from this study demonstrated that polymeric architecture and dendritic core structures play a crucial role in the determination of glass transition behavior, providing a strategy for the systematic engineering of polymer chain mobility.

Inhibition of Hepatitis C Virus (HCV) Replication by Hammerhead Ribozyme Which Activity Can Be Allosterically Regulated by HCV NS5B RNA Replicase (C형 간염바이러스(HCV)의 NS5B RNA Replicase에 의해 활성이 유도되는 Hammerhead 리보자임에 의한 HCV 복제 억제 연구)

  • Lee, Chang-Ho;Lee, Seong-Wook
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.188-193
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    • 2011
  • As a specific and effective therapeutic genetic material against hepatitis C virus (HCV) multiplication, HCV internal ribosome entry site (IRES)-targeting hammerhead ribozyme which activity is allosterically regulated by HCV regulatory protein, NS5B RNA replicase, was constructed. The allosteric ribozyme was composed of sequence of RNA aptamer to HCV NS5B, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding NS5B to the aptamer, and sequence of ribozyme targeting +382 nucleotide of HCV IRES. With real-time PCR analysis, the ribozyme was found to efficiently inhibit HCV replicon replication in cells. Of note, the allosteric ribozyme was shown to inhibit HCV replicon replication more efficiently than either HCV genome-targeting ribozyme or NS5B aptamer only. This allosteric ribozyme can be used as a lead genetic agent for the specific and effective suppression of HCV replication.