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A Role for Leu247 Residue within Transmembrane Domain 2 in Ginsenoside-Mediated α7 Nicotinic Acetylcholine Receptor Regulation

  • Lee, Byung-Hwan;Choi, Sun-Hye;Pyo, Mi Kyung;Shin, Tae-Joon;Hwang, Sung-Hee;Kim, Bo-Ra;Lee, Sang-MoK;Lee, Jun-Ho;Lee, Joon-Hee;Lee, Hui Sun;Choe, Han;Han, Kyou-Hoon;Kim, Hyoung-Chun;Rhim, Hyewhon;Yong, Joon-Hwan;Nah, Seung-Yeol
    • Molecules and Cells
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    • v.27 no.5
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    • pp.591-599
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    • 2009
  • Nicotinic acetylcholine receptors (nAChRs) play important roles in nervous system functions and are involved in a variety of diseases. We previously demonstrated that ginsenosides, the active ingredients of Panax ginseng, inhibit subsets of nAChR channel currents, but not ${\alpha}7$, expressed in Xenopus laevis oocytes. Mutation of the highly conserved Leu247 to Thr247 in the transmembrane domain 2 (TM2) channel pore region of ${\alpha}7$ nAChR induces alterations in channel gating properties and converts ${\alpha}7$ nAChR antagonists into agonists. In the present study, we assessed how point mutations in the Leu247 residue leading to various amino acids affect 20(S)-ginsenoside $Rg_3$ ($Rg_3$) activity against the ${\alpha}7$ nAChR. Mutation of L247 to L247A, L247D, L247E, L247I, L247S, and L247T, but not L247K, rendered mutant receptors sensitive to $Rg_3$. We further characterized $Rg_3$ regulation of L247T receptors. We found that $Rg_3$ inhibition of mutant ${\alpha}7$ nAChR channel currents was reversible and concentration-dependent. $Rg_3$ inhibition was strongly voltage-dependent and noncompetitive manner. These results indicate that the interaction between $Rg_3$ and mutant receptors might differ from its interaction with the wild-type receptor. To identify differences in $Rg_3$ interactions between wild-type and L247T receptors, we utilized docked modeling. This modeling revealed that $Rg_3$ forms hydrogen bonds with amino acids, such as Ser240 of subunit I and Thr244 of subunit II and V at the channel pore, whereas $Rg_3$ localizes at the interface of the two wild-type receptor subunits. These results indicate that mutation of Leu247 to Thr247 induces conformational changes in the wild-type receptor and provides a binding pocket for $Rg_3$ at the channel pore.

Analysis of Amperometric Response to Cholesterol according to Enzyme-Immobilization Methods (효소고정화 방법에 따른 콜레스테롤 검출용 바이오센서의 전류 감응도 분석)

  • Ji, Jung-Youn;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.21 no.5
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    • pp.731-738
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    • 2011
  • Cholesterol is the precursor of various steroid hormones, bile acid, and vitamin D with functions related to regulation of membrane permeability and fluidity. However, the presence of excess blood cholesterol may lead to arteriosclerosis and hypertension. Moreover, dietary cholesterol may affect blood cholesterol levels. Generally, cholesterol determination is performed by spectrophotometric or chromatographic methods, but these methods are very time consuming and costly, and require complicated pretreatment. Thus, the development of a rapid and simple analysis method for measuring cholesterol concentration in food is needed. Multi-walled carbon nanotube (MWCNT) was functionalized to MWCNT-$NH_2$ via MWCNT-COOH to have high sensitivity to $H_2O_2$. The fabricated MWCNT-$NH_2$ was attached to a glassy carbon electrode (GCE), after which Prussian blue (PB) was coated onto MWCNT-$NH_2$/GCE. MWCNT-$NH_2$/PB/GCE was used as a working electrode. An Ag/AgCl electrode and Pt wire were used as a reference electrode and counter electrode, respectively. The sensitivity of the modified working electrode was determined based on the amount of current according to the concentration of $H_2O_2$. The response increased with an increase of $H_2O_2$ concentration in the range of 0.5~500 ${\mu}M$ ($r^2$=0.96) with a detection limit of 0.1 ${\mu}M$. Cholesterol oxidase was immobilized to aminopropyl glass beads, CNBr-activated sepharose, Na-alginate, and toyopearl beads. The immobilized enzyme reactors with aminopropyl glass beads and CNBr-activated sepharose showed linearity in the range of 1~100 ${\mu}M$ cholesterol. Na-alginate and toyopearl beads showed linearity in the range of 5~50 and 1~50 ${\mu}M$ cholesterol, respectively. The detection limit of all immobilized enzyme reactors was 1 ${\mu}M$. These enzyme reactors showed high sensitivity; especially, the enzyme reactors with CNBr-activated sepharose and Na-alginate indicated high coupling efficiency and sensitivity. Therefore, both of the enzyme reactors are more suitable for a cholesterol biosensor system.

A Comparison of the Movement of Aspheric RGP Lens on Cornea by the Amounts of Keratometric Astigmatisms using Keratometer and Corneal Topography (각막곡률계와 각막지형도를 이용한 각막난시 측정값에 따른 비구면 RGP 렌즈의 각막에서 동적움직임 비교)

  • Park, Sang-Il;Lee, Se Eun;Kim, So Ra;Park, Mijung
    • Journal of Korean Ophthalmic Optics Society
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    • v.16 no.2
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    • pp.123-133
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    • 2011
  • Purpose: The present study was conducted to analyze any difference in the movement of aspheric RGP lens by the amounts of keratometiric astigmatisms using keratometer and corneal topography. Methods: Corneal curvatures in thirty six eyes of males and females of with-the-rule keratometric astigmatisms in their twenties were measured by a keratometer and worn aspheric RGP lenses. Then, lens rotations, vertical and horizontal movements of lens by blinking were measured to compare with lens movements when aspheric RGP lenses were fitted by total keratometric astigmatisms using corneal topography. Results: The case having higher amount of central keratometric astigmatism was 61.1% of subjects, however, 36.1% of subjects showed higher total keratometric astigmatism indicating that central keratometric astigmatism was not always bigger than total keratometric astigmatism. Since over 0.25 diopter difference between total and central keratometric astigmatisms was shown in 19 eyes (52.8% of subjects), the prescription for lens fitting could be changed. Significant difference in horizontal movement was detected with increase of astigmatism when it compared based on the amount of keratometric astigmatism measured by a keratometer. However, there was no significant difference in lens rotation, horizontal and vertical movements by comparison with the amount of total keratometric astigmatism using a corneal topography. When central keratometric astigmatism measured by keratometer was bigger than total keratometric astigmatism estimated by corneal topography, bigger lens rotation was shown compared with opposite case. Also, the tendency of bigger lens rotation was measured with the increase of keratomatric astigmatism in the case of same prescription having same base curves with same amount of keratometric astigmatism but different curvatures. Conclusions: From the present study, we concluded that lens movements on cornea were not totally different when aspheric RGP lens fitted on with-the-rule astigmatism by keratometer and corneal topography. However, there was some difference in certain lens movements. Therefore, we concluded that further study on the relationship between the prescriptions for lens fitting should be conducted for improving the rate of successful lens fitting by keratometer or for the proper application of corneal topography for lens fitting.

Antithrombotic Activities of Cheongkookjang and Cheongkookjang Fermented with Green Tea or Mugwort (청국장 및 녹차, 쑥이 첨가된 청국장의 항혈전 활성)

  • Lee, Kyung-Ae;Jang, Jeong-Oak;Yoon, Hye-Kyung;Kim, Moo-Sung
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.298-303
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    • 2007
  • Antithrombotic activities of water extract of cheongkookjang and cheongkookjang fermented with green tea or mugwort were evaluated on some antithrombosis related activities in vitro and thrombotic death inhibition in vivo. Cheongkookjang made of white soybean (Glycine max) or black small soybean (Rhynchosia nulubilis) showed potent antioxidative activities. Addition of green tea or mugwort during cheongkookjang fermentation increased the antioxidative activity, cheongkookjang with green tea showed more drastic increase compared with cheongkookjang with mugwort. Nitrite scavenging effects of the cheongkookjang extracts were prominent but the addition of green tea or mugwort seldom increased the scavenging effects. All the cheongkookjang extracts showed strong inhibitory activities on platelet aggregation. The inhibitory activities of cheongkookjang were increased considerably by addition of green tea or mugwort even with low concentration. Plasmin unit as fibrinolytic activity was not affected considerably by addition of green tea. Addition of mugwort decreased the activity transiently at low concentration ($0.3{\sim}1.0%$) but increased again slowly at higher concentration ($1{\sim}3%$). In vitro thrombotic death inhibition test, the antithrombotic activity of cheongkookjang made of black small bean with green tea was higher by about 1.5 times compared to that without green tea. As results, cheongkookjang might inhibit antithrombosis not only by fibrinolytic action but also by inhibition of platelet aggregation and antioxidative action. The addition of functional materials such as green tea or mugwort could increase the antithrombotic function, even at low concentration.

A Development and Application of the Landscape Evaluation Model Based on the Biotope Classification (비오톱 유형분류를 기반으로 한 경관평가 모형개발 및 적용)

  • Park, Cheon-Jin;Ra, Jung-Hwa;Cho, Hyun-Ju;Kim, Jin-Hyo;Kwon, Oh-Sung
    • Journal of the Korean Institute of Landscape Architecture
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    • v.40 no.4
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    • pp.114-126
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    • 2012
  • The purpose of this study is to find ways of the view evaluation of biotope classification before development by selecting an area, which is as large as about $10.0km^2$ around Non Gong Up, Auk Po Myun, Dalsung Gun, Daugu where the large project has been planned, as a subject of this study. The results of this study are as follows. Because of the classification of biotope, there are 23 kinds of types that are subdivided into 140 types. Three surveys for selecting the assessment indicators were performed. The first survey analyzed the importance of 22 selected assessment indicators based on the evaluation of an existing literature review and on the spot research. The second survey performed factor analysis and reclassified the value indicators. The third survey computed additive values of the selected assessment indicators. It used a method of standardizing the average importance of indicators by making their sum equal by 10. Theses additive values were then multiplied by each grade of indicators in order to make a final evaluation. The number of assessment indicators finally selected through the survey of asking specialist is vitality elements, visual obstructs elements etc 19. According to the result of evaluation of 1st, 1 grade spaces which especially valuable is analyzed that 7 spaces, 2 grade spaces for 4, 3 grade spaces for 5, 4 grade space for 2, 5 grade space for 5. Because of the evaluation of 2st, 1 grade spaces which especially valuable(1a, 1b) is analyzed that 15 spaces, 2 grade spaces which valuable is analyzed that 28 space. As the evaluation of site suitability model of this study couldn't have high applicability to other similar area because of having only one site as a subject, it is needed to do synthesize and standardization of various examples to have higher objectivity later.

Effects of Temperature and Fluctuation Range on Microbial Growth and Quality of Foods Stored in Domestic Refrigerator (냉장실의 온도 정온화가 냉장 식품의 품질과 미생물의 생육에 미치는 영향)

  • Jung, Dong-Sun;Kweon, Mee-Ra;Auh, Joong-Hyuck;Cho, Kwang-Yeun;Choi, Young-Hoon;Kook, Seung-Uk;Park, Kwan-Hwa
    • Korean Journal of Food Science and Technology
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    • v.28 no.4
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    • pp.632-637
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    • 1996
  • Effects of refrigeration temperature and its fluctuation range on the growth of psychrotrophic microorganisms and the quality of refrigerated foods such as apple, fish and oyster were evaluated to find optimum storage conditions for a domestic refrigerator. Refrigeration temperature was $2^{\circ}C$ or $4^{\circ}C$, and fluctuation ranges were varied: ${\pm}0.3,\;{\pm}1.0,\;{\pm}1.2,\;or\;{\pm}4.0^{\circ}C$. Changes in hardness of apples stored at $2{\pm}0.3^{\circ}C$ were much slower than those of apples stored at $4{\pm}1.2^{\circ}C$. Freshness of fish and oyster also lasted much longer at low temperature such as $2{\pm}0.3^{\circ}C$. The growth of Listeria monocytogenes inoculated on sliced ham was inhibited for 1 month at $2{\pm}0.3^{\circ}C$, but the cells at $4{\pm}1.2^{\circ}C$ began to grows as time elapsed. Therefore, it was expected that shelf-life of certain food stored in a domestic refrigerator could be extended by lowering temperature to $2^{\circ}C$ and by reducing fluctuation range of refrigerator.

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An International Collaborative Program To Discover New Drugs from Tropical Biodiversity of Vietnam and Laos

  • Soejarto, Djaja D.;Pezzuto, John M.;Fong, Harry H.S.;Tan, Ghee Teng;Zhang, Hong Jie;Tamez, Pamela;Aydogmus, Zeynep;Chien, Nguyen Quyet;Franzblau, Scott G.;Gyllenhaal, Charlotte;Regalado, Jacinto C.;Hung, Nguyen Van;Hoang, Vu Dinh;Hiep, Nguyen Tien;Xuan, Le Thi;Hai, Nong Van;Cuong, Nguyen Manh;Bich, Truong Quang;Loc, Phan Ke;Vu, Bui Minh;Southavong, Boun Hoong
    • Natural Product Sciences
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    • v.8 no.1
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    • pp.1-15
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    • 2002
  • An International Cooperative Biodiversity Group (ICBG) program based at the University of Illinois at Chicago initiated its activities in 1998, with the following specific objectives: (a) inventory and conservation of of plants of Cuc Phuong National Park in Vietnam and of medicinal plants of Laos; (b) drug discovery (and development) based on plants of Vietnam and Laos; and (c) economic development of communities participating in the ICBG project both in Vietnam and Laos. Member-institutions and an industrial partner of this ICBG are bound by a Memorandum of Agreement that recognizes property and intellectual property rights, prior informed consent for access to genetic resources and to indigenous knowledge, the sharing of benefits that may arise from the drug discovery effort, and the provision of short-term and long-term benefits to host country institutions and communities. The drug discovery effort is targeted to the search for agents for therapies against malaria (antimalarial assay of plant extracts, using Plasmodium falciparum clones), AIDS (anti-HIV-l activity using HOG.R5 reporter cell line (through transactivation of the green fluorescent protein/GFP gene), cancer (screening of plant extracts in 6 human tumor cell lines - KB, Col-2, LU-l, LNCaP, HUVEC, hTert-RPEl), tuberculosis (screening of extracts in the microplate Alamar Blue assay against Mycobacterium tuberculosis $H_{37}Ra\;and\;H_{37}Rv),$ all performed at UIC, and CNS-related diseases (with special focus on Alzheimer's disease, pain and rheumatoid arthritis, and asthma), peformed at Glaxo Smith Kline (UK). Source plants were selected based on two approaches: biodiversity-based (plants of Cuc Phuong National Park) and ethnobotany-based (medicinal plants of Cuc Phuong National Park in Vietnam and medicinal plants of Laos). At mc, as of July, 2001, active leads had been identified in the anti-HIV, anticancer, antimalarial, and anti- TB assay, after the screening of more than 800 extracts. At least 25 biologically active compounds have been isolated, 13 of which are new with anti-HIV activity, and 3 also new with antimalarial activity. At GSK of 21 plant samples with a history of use to treat CNS-related diseases tested to date, a number showed activity against one or more of the CNS assay targets used, but no new compounds have been isolated. The results of the drug discovery effort to date indicate that tropical plant diversity of Vietnam and Laos unquestionably harbors biologically active chemical entities, which, through further research, may eventually yield candidates for drug development. Although the substantial monetary benefit of the drug discovery process (royalties) is a long way off, the UIC ICBG program provides direct and real-term benefits to host country institutions and communities.

Cell study on the Magnesium ion implanted surface with PSII (PSII를 이용한 마그네슘 이온 주입 임플란트에 대한 MC3T3-E1 골모양 세포 반응 연구)

  • Shin, Hyeong-Joo;Kim, Dae-Gon;Park, Chan-Jin;Cho, Lee-Ra;Lee, Hee-Su;Cha, Min-Sang
    • Journal of Dental Rehabilitation and Applied Science
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    • v.25 no.4
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    • pp.361-374
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    • 2009
  • For successful osteogenesis around the implants, interaction between implant surface and surrounding tissue is important. Biomechanical bonding and biochemical bonding are considered to influence the response of adherent cells. But the focus has shifted surface chemistry. The purpose of this study is to evaluate the MC3T3-E1 osteoblast like cell responses of magnesium (Mg) ion implanted titanium surface produced using a plasma source ion implantation method. Commercially pure titanium disc was used as substrates. The discs were prepared to produce four different surface, A: Machine turned surface, B: Mg implanted surface, C: sandblasted surface, D: sandblasted and Mg implanted surface. MC3T3 El osteoblastic like cells were cultured on the disc specimens. Cell adhesion, proliferation, differentiation, and synthesis of extracellular matrix were evaluated. The cell adhesion morphology was evaluated by SEM. RT PCR assay was used for assessment of cell adhesion, proliferation and differentiation. ALP activity was measured for cell differentiation. The results of this study were as follows: 1. SEM showed that cell on Mg ion groups was more proliferative than that of non Mg ion groups. On the machine turned surface, cell showed some degree of contact guidance in aligning with the machining grooves. 2. In RT PCR analysis, osteonectin and c-fos mRNA were more expressed on sandblasted and Mg ion implanted group. 3. ALP activity was not significantly different among all groups. Within the limitations of this study, the following conclusions were drawn: It might indicate Mg ion implanted titanium surface induce better bone response than non Mg ion groups.

Flow Behavior and Mixing Characteristics of Rice Husk/Silica Sand/Rice Husk Ash (왕겨/모래/왕겨 회재의 유동 및 혼합 특성 연구)

  • Kim, Bo Hwa;Seo, Myung Won;Kook, Jin Woo;Choi, Hee Mang;Ra, Ho Won;Yoon, Sang Jun;Mun, Tae Young;Kim, Yong Ku;Lee, Jae Goo;Rhee, Young Woo
    • Korean Chemical Engineering Research
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    • v.54 no.4
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    • pp.533-542
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    • 2016
  • We investigate fluidization characteristics of the mixture of rice husk, silica sand and rice husk ash as a preliminary study for valuable utilization of rice husk ash obtained from gasification of rice husk in a fluidized bed reactor. As experiment valuables, the blending ratio of rice husk and sand (rice husk: sand) is selected as 5:95, 10:90, 20:80 and 30:70 on a volume base. Rice husk ash was added with 6 vol% of rice husk for each experiment and air velocity to the reactor was 0~0.63 m/s. In both rice husk/sand and rice husk/sand/ash mixture, the minimum fluidization velocity (Umf) is observed as 0.19~0.21 m/s at feeding of 0~10 vol.% of rice husk and 0.30 m/s at feeding of 20 vol.% of rice husk. With increasing the amount of rice husk up to 30 vol.%, $U_{mf}$ can not measure due to segregation behavior. The mixing index for each experiment is determined using mixing index equation proposed by Brereton and Grace. The mixing index of the mixture of rice husk/sand and rice husk/sand/ash was 0.8~1 and 0.88~1, respectively. The optimum fluidization condition was found for the good mixing and separation of rice husk ash.

Effects of Feeding Level of Extruded Poultry Manure on Growth Performance, Nutrient Digestibility and Body Composition in Korean Native Goats (Extrusion 가공처리 계분사료의 첨가수준이 재래산양의 육성능력, 영양소 소화율 및 체조성에 미치는 영향)

  • Kim, C.H.;Ra, C.S.;Goh, B.D.;Park, J.I.;Lin, G.Z.;Shin, J.S.
    • Journal of Animal Science and Technology
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    • v.44 no.6
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    • pp.783-792
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    • 2002
  • This experiment was conducted to investigate the effects of supplementary level of extruded poultry manure, corn and tapioca mixture (EPM) on growth performance, nutrient digestibility and body composition in korean native goats. Total 15 heads of Korean native goats (10 kg B.W.) were randomly assigned into five treatment groups (EPM 10, 20, 30, 40% and control.) and feeding trial was done for six weeks with ad libitum. Protein level of the extrudate with poultry manure was directly proportional to corn supplements rate. NFE and Ca content in EPM also were functional of the tapioca supplements levels. Daily body weight gain was significantly (p<0.05) reduced in EPM 40% group, but no significant differences were observed in other groups. Concentrate feed intake was high in EPM 10 to 30% groups, compared to the control. Feed conversion ratio also was relatively higher in EPM groups than in the control. As EPM level increased, dry matter digestibility was proportionally decreased, but that of crude protein was enhanced. Dressed carcass percentage was significantly (p<0.05) reduced with the increase of EPM level, showing control 49.7, EPM 10% 49.8, EPM 20% 48.3, EPM 30% 47.9 and EPM 40% 45.2, respectively.