• Title/Summary/Keyword: Molecular Characteristics

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Functional Characteristics of TRPC4 Channels Expressed in HEK 293 Cells

  • Sung, Tae Sik;Kim, Min Ji;Hong, Soojin;Jeon, Jae-Pyo;Kim, Byung Joo;Jeon, Ju-Hong;Kim, Seon Jeong;So, Insuk
    • Molecules and Cells
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    • v.27 no.2
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    • pp.167-173
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    • 2009
  • The classical type of transient receptor potential (TRPC) channel is a molecular candidate for $Ca^{2+}$-permeable cation channels in mammalian cells. Because TRPC4 and TRPC5 belong to the same subfamily of TRPC, they have been assumed to have the same physiological properties. However, we found that TRPC4 had its own functional characteristics different from those of TRPC5. TRPC4 channels had no constitutive activity and were activated by muscarinic stimulation only when a muscarinic receptor was co-expressed with TRPC4 in human embryonic kidney (HEK) cells. Endogenous muscarinic receptor appeared not to interact with TRPC4. TPRC4 activation by $GTP{\gamma}S$ was not desensitized. TPRC4 activation by $GTP{\gamma}S$ was not inhibited by either Rho kinase inhibitor or MLCK inhibitor. TRPC4 was sensitive to external pH with $pK_a$ of 7.3. Finally, TPRC4 activation by $GTP{\gamma}S$ was inhibited by the calmodulin inhibitor W-7. We conclude that TRPC4 and TRPC5 have different properties and their own physiological roles.

Monolayer Characteristics of Bilayer Forming Phosphate Amphiphiles (이분자막 형성능을 가지는 인산형 양친매성 화합물의 단분자막 특성)

  • ;Kunitake, T.
    • Membrane Journal
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    • v.5 no.2
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    • pp.89-96
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    • 1995
  • The monolayer characteristics of phosphate amphiphiles with azobenzene at air/water interface were studied by the measurment of $\pi-A$ curves and absorption spectra. Immediately after being spread on the water surface, these amphiphiles having strong intermolecular hydrogen bonding interactions showed the typical absorption spectra which resulted from domain formation. But the aggregated domains could be controlled by changing the subphase conditions (adding bulky salt and rasing pH). Addition of metal ions in subphase changes the molecular orientation of monolayer. As the metal ion charge increases ($1\leq2$ < 3 < 4 valence), the absorption maximum (310nm) of the amphiphile with azobenzene shifts to a longer wavelength (350nm) which means that the orientation of the amphiphile is tilted. These results suggest that the molecular orientation, and furthermore the aggregation state of monolayer can be possibly controlled by the interaction of metal ions with different charge types.

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Construction of Artificial Epithelial Tissues Prepared from Human Normal Fibroblasts and C9 Cervical Epithelial Cancer Cells Carrying Human Papillomavirus Type 18 Genes

  • Eun Kyung Yang;Seu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.1
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    • pp.1-5
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    • 1998
  • One cervical cancer cell line, C9, carrying human papillomavirus type 18 (HPV18) genes that is one of the major etiologic concoviruses for cervical cancer was characterized. This cell line was further characterized for its capacity related to the epithelial cell proliferation, stratification and differentiation in reconstituted artificial epithelial tissue. The in vitro construction of three dimensional artificial cervical opithelial tissue has been engineered using C9 epithelial cancer cells, human foreskin fibroblasts and a matrix made of type I collagen by organotypic culture of epithelial cells. The morphology of paraffin embedded artificial tissue was examined by histochemical staining. The artificial epithelial tissues were well developed having multilayer. However, the tissue morphology was similar to the cervical tissus having displasia induced by HPV infection. The characteristics of the artificial tissues were examined by determinining the expression of specific marker proteins. In the C9 derived artificial tissues, the expression of EGF receptor, as epithelial proliferation marker proteins for stratum basale was observed up to the stratum spinosum. Another epithelial proliferation marker for stratum spinosum, cytokerations 5/6/18, were observed well over the stratum spinosum. For the differentiation markers, the expression of involucrin and filaggrin were observed while the terminal differentiation marker, cytokeratins 10/13 was not detected at all. Therefore the reconstituted artificial epithelial tissues expressed the same types of differentiation marker proteins that are expressed in normal human cervical epithelial tissues but lacked the final differentiation capacity representing characteristics of C9 cell line as a cancer tissue devived cell line. Expression of HPV18 E6 oncoprotein was also observed in this artifical cervical opithelial tissue though the intensity of the staining was weak. Thus this artificial epithelial tissue could be used as a useful model system to examine the relationship between HPV-induced cervical oncogenesis and epithelial cell differentiation.

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A Study on the Current-Voltage Characteristics of Self-Assembled Nitro-group and Methoxy-group Organic Molecules by Using STM (STM을 이용한 자기조립된 니트로기와 메톡시기 유기분자의 전압-전류 특성 연구)

  • Kim, Seung-Un;Park, Sang-Hyun;Park, Jae-Chul;Shin, Hoon-Kyu;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2004.11a
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    • pp.212-214
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    • 2004
  • In this study, we fabricated the organic thin film by self-assembly method by using nitro-group and methoxy-group organic molecule. Also, we selected the organic single molecule in organic thin film and measured current-voltage characteristics by using scanning tunneling microscopy. The Organic molecules that use in an experiment is 4,4'-(diethynylphenyl)-2'-nitro-1-benzen ethiol and 4-[2,5-dimethoxy-4-ph enylethynylphenyl]ethynylphenylethanthiol. 4,4'-(dimet hynylphenyl)-2'-nitro-1-benzenethiol is applied widely in molecular electronic device and 4-[2,5-dime thoxy-4-phenylethynylphenyl]ethynylphenylethanthiol composed in Korea Research Institute of Chemical Technology. To be confirmed the formation of the self-assembled monolayers, we observed the real time frequency shift of the QCM and investigated surface of the self-assembled monolayers the using STM. With this, we measured current to the organic single molecule, in condition of the air state. As a result, we confirmed in constant voltage that properties of negative differential resistance. Using properties of negative differential resistance to get from this study, application is expected to be molecular switching device, memory device and logic device.

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The First Case of Diarrhoea in Tibetan Sheep, Ovis aries, Caused by Balantidium coli in the Qinghai Tibetan Plateau Area, China

  • Jian, Ying-Na;Wang, Ge-Ping;Li, Xiu-Ping;Zhang, Xue-Yong;Ma, Li-Qing
    • Parasites, Hosts and Diseases
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    • v.56 no.6
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    • pp.603-607
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    • 2018
  • This study was carried out to determine the pathogen-causing diarrhoea in sheep Ovis aries in the Qinghai Tibetan Plateau Area, China. A trophozoite was identified as species of ciliate alveolates infecting the sheep based on morphological characteristics examined by microscope. It was mostly spherical, colourless and transparent, with many vesicles. Macronucleus and contractile vacuoles could not be distinguished. Size of the trophozoite was $80-180{\times}70-150{\mu}m$ and its surface was covered with cilia. Molecular analysis based on sequences of 18S rRNA and ITS genes confirmed the ciliate species as Balantidium coli. According to the literature, there have been many epidemiological investigations of B. coli infection in pigs, monkeys and humans. To our knowledge, this was the first report of B. coli infections in sheep in the Qinghai Tibetan Plateau Area of China, or eleswhere around the world. Importantly, the sheep case was rare but raised our concern that B. coli may spread across species and expand its host range.

Technology Trend on Commercial Polymeric Membranes for Water Treatment (수처리용 상용 고분자 분리막 제품 기술동향)

  • Jang, Haenam
    • Membrane Journal
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    • v.29 no.1
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    • pp.11-17
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    • 2019
  • In the field of water treatment membranes, polymers are used together with ceramics as the most important materials. In this review, I tried to analyze the technology trends of polymer membrane materials based on commercial products. For this purpose, according to the types of water treatment membranes such as MF (Microfiltration), UF (Ultrafiltration) and NF (Nanofiltration), the trends of polymer membrane products were investigated by countries, materials, and companies. Through this, we were able to classify the types of materials that are mainly used for each type of membrane, and at the same time, identify the companies that are dominant in the market, and analyze which materials constitute the product portfolio. Based on these results, we have presented the characteristics of the material market according to each type of membrane, and proposed a technology development strategy to enter each market based on these characteristics.

Increased mRNA Stability and Expression Level of Croceibacter atlanticus Lipase Gene Developed through Molecular Evolution Process

  • Jeong, Han Byeol;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.6
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    • pp.882-889
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    • 2021
  • In order to use an enzyme industrially, it is necessary to increase the activity of the enzyme and optimize the reaction characteristics through molecular evolution techniques. We used the error-prone PCR method to improve the reaction characteristics of LipCA lipase discovered in Antarctic Croceibacter atlanticus. Recombinant Escherichia coli colonies showing large halo zones were selected in tributyrin-containing medium. The lipase activity of one mutant strain (M3-1) was significantly increased, compared to the wild-type (WT) strain. M3-1 strain produced about three times more lipase enzyme than did WT strain. After confirming the nucleotide sequence of the M3-1 gene to be different from that of the WT gene by four bases (73, 381, 756, and 822), the secondary structures of WT and M3-1 mRNA were predicted and compared by RNAfold web program. Compared to the mean free energy (MFE) of WT mRNA, that of M3-1 mRNA was lowered by 4.4 kcal/mol, and the MFE value was significantly lowered by mutations of bases 73 and 756. Site-directed mutagenesis was performed to find out which of the four base mutations actually affected the enzyme expression level. Among them, one mutant enzyme production decreased as WT enzyme production when the base 73 was changed (T→ C). These results show that one base change at position 73 can significantly affect protein expression level, and demonstrate that changing the mRNA sequence can increase the stability of mRNA, and can increase the production of foreign protein in E. coli.

Biological Characteristics of Recombinant Arthrobotrys oligospora Chitinase AO-801

  • Gong, Shasha;Meng, Qingling;Qiao, Jun;Huang, Yunfu;Zhong, Wenqiang;Zhang, Guowu;Zhang, Kai;Li, Ningxing;Shang, Yunxia;Li, Zhiyuan;Cai, Xuepeng
    • Parasites, Hosts and Diseases
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    • v.60 no.5
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    • pp.345-352
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    • 2022
  • Chitinase AO-801 is a hydrolase secreted by Arthrobotrys oligospora during nematode feeding, while its role remained elusive. This study analyzed the molecular characteristics of recombinant chitinase of Arthrobotrys oligospora (reAO-801). AO-801 belongs to the typical glycoside hydrolase 18 family with conserved chitinase sequence and tertiary structure of (α/β)8 triose-phosphate isomerase (TIM) barrel. The molecular weight of reAO-801 was 42 kDa. reAO-801 effectively degraded colloidal and powdered chitin, egg lysate, and stage I larval lysate of Caenorhabditis elegans. The activity of reAO-801 reached its peak at 40℃ and pH values between 4-7. Enzyme activity was inhibited by Zn2+, Ca2+, and Fe3+, whereas Mg2+ and K+ potentiated its activity. In addition, urea, sodium dodecyl sulfate, and 2-mercaptoethanol significantly inhibited enzyme activity. reAO-801 showed complete nematicidal activity against C. elegans stage I larvae. reAO-801 broke down the C. elegans egg shells, causing them to die or die prematurely by hatching the eggs. It also invoked degradation of Haemonchus contortus eggs, resulting in apparent changes in the morphological structure. This study demonstrated the cytotoxic effect of reAO-801, which laid the foundation for further dissecting the mechanism of nematode infestation by A. oligospora.

Molecular Characteristics and Potent Immunomodulatory Activity of Fasciola hepatica Cystatin

  • Zhang, Kai;Liu, Yucheng;Zhang, Guowu;Wang, Xifeng;Li, Zhiyuan;Shang, Yunxia;Ning, Chengcheng;Ji, Chunhui;Cai, Xuepeng;Xia, Xianzhu;Qiao, Jun;Meng, Qingling
    • Parasites, Hosts and Diseases
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    • v.60 no.2
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    • pp.117-126
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    • 2022
  • Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-α, and promoted the expression of transforming growth factor-β and interleukin-10. These results showed that FhCystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection.

Changes in Spectroscopic and Molecular Weight Characteristics of Dissolved Organic Matter in an Agriculture Reservoir during a Summer Monsoon (장마시기에 따른 농업용 저수지 내 용존 유기물 분광특성과 분자량 변화)

  • Jung, Ka-Young;Lee, Yun Kyung;Yoo, HaYoung;Nam, Gui-Sook;Hur, Jin
    • Journal of Korean Society on Water Environment
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    • v.37 no.6
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    • pp.458-468
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    • 2021
  • In this study, we investigated the variations of dissolved organic matter (DOM) in an agricultural reservoir during the monsoon period (June to October, 2020) with respect to the organic carbon concentration (DOC), molecular weight distribution, and optical properties. The monsoon period was divided into three phases - beginning storm (BS), during storm (DS), and after storm (AS). Our results showed significant differences in the concentrations and characteristics of DOM during the summer monsoon. The DOC concentrations were decreased after the monsoon, probably due to a dilution effect. In contrast, increasing trends were observed in the specific UV absorbance (SUVA), and relative abundances of humic-like fluorescence and larger-sized compounds. These observations implied that the large-sized and humic-like organic components with terrestrial origins strongly affected the reservoir DOM after the summer monsoon. Meanwhile, biopolymer size fraction, which is associated with algal activity, became more abundant after the monsoon. These results suggest that DOM with autochthonous sources became dominant as a result of the inflow of nutrients into the reservoir after the storm. Spatial changes in DOM within the reservoir were not pronounced as much as the temporal variations. All taken, it can be concluded that the summer monsoon simply led to the decrease of DOM concentrations while the sources and the quality of DOM underwent substantial changes, which may enrich refractory organic matter in the reservoir. This study reveals the importance of in-depth DOM quality monitoring before and after summer monsoon for effective water quality management in agricultural reservoirs.