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Differential Effects of Quercetin and Quercetin Glycosides on Human α7 Nicotinic Acetylcholine Receptor-Mediated Ion Currents

  • Lee, Byung-Hwan;Choi, Sun-Hye;Kim, Hyeon-Joong;Jung, Seok-Won;Hwang, Sung-Hee;Pyo, Mi-Kyung;Rhim, Hyewhon;Kim, Hyoung-Chun;Kim, Ho-Kyoung;Lee, Sang-Mok;Nah, Seung-Yeol
    • Biomolecules & Therapeutics
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    • v.24 no.4
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    • pp.410-417
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    • 2016
  • Quercetin is a flavonoid usually found in fruits and vegetables. Aside from its antioxidative effects, quercetin, like other flavonoids, has a various neuropharmacological actions. Quercetin-3-O-rhamnoside (Rham1), quercetin-3-O-rutinoside (Rutin), and quercetin-3-(2(G)-rhamnosylrutinoside (Rham2) are mono-, di-, and tri-glycosylated forms of quercetin, respectively. In a previous study, we showed that quercetin can enhance ${\alpha}7$ nicotinic acetylcholine receptor (${\alpha}7$ nAChR)-mediated ion currents. However, the role of the carbohydrates attached to quercetin in the regulation of ${\alpha}7$ nAChR channel activity has not been determined. In the present study, we investigated the effects of quercetin glycosides on the acetylcholine induced peak inward current ($I_{ACh}$) in Xenopus oocytes expressing the ${\alpha}7$ nAChR. $I_{ACh}$ was measured with a two-electrode voltage clamp technique. In oocytes injected with ${\alpha}7$ nAChR copy RNA, quercetin enhanced $I_{ACh}$, whereas quercetin glycosides inhibited $I_{ACh}$. Quercetin glycosides mediated an inhibition of $I_{ACh}$, which increased when they were pre-applied and the inhibitory effects were concentration dependent. The order of $I_{ACh}$ inhibition by quercetin glycosides was Rutin${\geq}$Rham1>Rham2. Quercetin glycosides-mediated $I_{ACh}$ enhancement was not affected by ACh concentration and appeared voltage-independent. Furthermore, quercetin-mediated $I_{ACh}$ inhibition can be attenuated when quercetin is co-applied with Rham1 and Rutin, indicating that quercetin glycosides could interfere with quercetin-mediated ${\alpha}7$ nAChR regulation and that the number of carbohydrates in the quercetin glycoside plays a key role in the interruption of quercetin action. These results show that quercetin and quercetin glycosides regulate the ${\alpha}7$ nAChR in a differential manner.

Characterization of a Small Cryptic Plasmid from Pseudomonas nitroreducens Strain TX1 (Pseudomonas nitroreducens TX1에 존재하는 작은 플라스미드의 특성 규명)

  • Nguyen, Ngoc Tuan;Lee, Kyoung;Kang, Ju Beom;Huang, Shir-Ly
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.210-215
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    • 2014
  • Pseudomonas nitroreducens TX1 was isolated from a rice field drainage in Taiwan. The bacterium is of special interest because of its capability to use a group of nonionic surfactants such as alkylphenol polyethoxylates even at high concentrations as a sole carbon source. In this study, a small cryptic circular plasmid, pTX1, was characterized from P. nitroreducens TX1. It is 2,286 bp in length with a GC content of 63.3% and harbors three open reading frames, $Rep_{pTX1}$ and functionally unidentified ORF1 and ORF2. The predicted $rep_{pTX1}$ gene product is homologous to Rep proteins of plasmids belonging to the pC194/pUB110 family, which is predominantly found in Gram-positive bacteria and is known to replicate by the rolling-circle mechanism. The copy number of pTX1 was estimated to be about 150 in each cell. Based on the genetic fingerprints and comparison with other plasmids, it is concluded that pTX1 replicates by a rolling circle mechanism which is rarely found for Pseudomonas plasmids.

Environmental risk assessment of genetically modified Herbicide-Tolerant zoysiagrass (Event: Jeju Green21) (제초제저항성 들잔디(Zoysia japonica Steud.) 이벤트 Jeju Green21의 환경위해성평가)

  • Bae, Tae-Woong;Kang, Hong-Gyu;Song, In-Ja;Sun, Hyeon-Jin;Ko, Suk-Min;Song, Pill-Soon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.105-116
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    • 2011
  • Transgenic zoysiagrass (Zoysia japonica Steud.) expressing the bar gene inserted in the plant genome has been generated previously through Agrobacterium tumefaciens-mediated transformation. The GM zoysiagrass (event: JG21) permits efficient management of weed control of widely cultivated zoysiagrass fields, reducing the frequency and cost of using various herbicides for weed control. Now we have carried out the environmental risk assessment of JG21 prior to applying to the governmental regulatory agency for the commercial release of the GM turf grass outside of test plots. The morphological phenotypes, molecular analysis, weediness and gene flow from each test plot of JG21 and wild-type zoysiagrasses have been evaluated by selectively analyzing environmental effects. There were no marked differences in morphological phenotypes between JG21 and wild-type grasses. The JG21 retained its stable integration in the host plant in T1 generation, exhibiting a 3:1 segregation ratio according to the Mendelian genetics. We confirmed the copy number (1) of JG21 by using Southern blot analysis, as the transgenic plants were tolerant to ammonium glufosinate throughout the culture period. From cross-fertilization and gene flow studies, we found a 9% cross-pollination rate at the center of JG21 field and 0% at distances over 3 m from the field. The JG21 and wild-type zoysiagrass plants are not considered "weed" because zoysiagrasses generally are not dominant and do not spread into weedy areas easily. We assessed the horizontal gene transfer (HGT) of the transgene DNA to soil microorganisms from JG21 and wild-type plants. The bar gene was not detected from the total genomic DNA extracted from each rhizosphere soil of GM and non-GM Zoysia grass fields. Through the monitoring of JG21 transgene's unintentional release into the environment, we found no evidence for either pollen mediated gene flow of zoysiagrass or seed dispersal from the test field within a 3 km radius of the natural habitat.

Proporties of SV4O-transformed Human Cells (SV40 바이러스로 형질전환된 사람종양세포의 특성)

  • 최경희;홍승환
    • The Korean Journal of Zoology
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    • v.31 no.1
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    • pp.49-55
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    • 1988
  • A few SV4O-transformed human cells such as SV8O are potentially tumorigenic but rejected by athymic hosts. However, one cell line in this group (W118IVA-2) is known to be fully tumorigenic. Two clones were obtained after the injection of W118IVA-2, of which NW1SC1-1 was tumorigenic but NW18C1-2 was not in nude mice. As examined by Southern blot analysis, NW18C1-1 appears to contain more copy number of SV40 sequences than NW18C1-2 does. However, it was unable to demonstrate that this difference elicits the tumorigenicity in NW18C1-1 but not in NW18C1-2. Therefore, the latter clone was tested if it expresses SV40 early genes to produce large T as well as small t antigens using indirect immunofluorescent assay and immunoprecipitation. In addition, mouse NIH3T3 cells were transfected with the cellular DNA of NW1SC1-2 as well as that of NW18C1-1 to examine if the viral genomes in the clones can make the nontransformed cells to acquire malignant growth potential in vivo. The transformed cells expressed large T antigen and became tumorigenic. Thus, the transforming functions of NW1SC1-2 cell appers to be intact. These results clearly suggest that the inability of NW18C1-2 cell to form tumor in nude mice is not because they are inherently nontumorigenic. However, the possibility that the interaction of SV40 with its host differs in these clones can not he ruled out.

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Expression of Toll-like Receptors, Pro-, and Anti-inflammatory Cytokines in Relation to Gut Microbiota in Irritable Bowel Syndrome: The Evidence for Its Micro-organic Basis

  • Shukla, Ratnakar;Ghoshal, Ujjala;Ranjan, Prabhat;Ghoshal, Uday C
    • Journal of Neurogastroenterology and Motility
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    • v.24 no.4
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    • pp.628-642
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    • 2018
  • Background/Aims A Subset of patients with irritable bowel syndrome (IBS) may have mild inflammation due to immune activation. Toll-like receptors (TLRs) and cytokines may cause intestinal inflammation. We studied their expression in relation to gut microbiota. Methods Expression of TLRs and cytokines was assessed in 47 IBS patients (Rome III) and 25 controls using quantitative real-time polymerase chain reaction. Immunohistochemistry was further performed to confirm the expression of TLR-4 and TLR-5. Results Of 47 patients with IBS, 20 had constipation (IBS-C), 20 diarrhea (IBS-D), and 7 unclassified (IBS-U). The mRNA levels of TLR-4 and TLR-5 were up-regulated in IBS patients than controls (P = 0.013 and P < 0.001, respectively). Expression of TLR-4 and TLR-5 at protein level was 4.2-folds and 6.6-folds higher in IBS-D than controls. The mRNA levels of IL-6 (P = 0.003), C-X-C motif chemokine ligand 11 (CXCL-11) (P < 0.001) and C-X-C motif chemokine receptor 3 (CXCR-3) (P < 0.001) were higher among IBS patients than controls. Expression of IL-6 (P = 0.002), CXCL-11 (P < 0.001), and CXCR-3 (P < 0.001) were up-regulated and IL-10 (P = 0.012) was down-regulated in IBS-D patients than controls. Positive correlation was seen between TLR-4 and IL-6 (P = 0.043), CXCR-3, and CXCL-11 (P = 0.047), and IL-6 and CXCR-3 (P = 0.003). Stool frequency per week showed positive correlation with mRNA levels of TLR-4 (P = 0.016) and CXCR-3 (P = 0.005), but inversely correlated with IL-10 (P = 0.002). Copy number of Lactobacillus (P = 0.045) and Bifidobacterium (P = 0.011) showed correlation with IL-10 in IBS-C, while Gram-positive (P = 0.031) and Gram-negative bacteria (P = 0.010) showed correlation with CXCL-11 in IBS-D patients. Conclusions Altered immune activation in response to dysbiotic microbiota may promote intestinal inflammation in a subset of patients with IBS.

Performance of Uncompressed Audio Distribution System over Ethernet with a L1/L2 Hybrid Switching Scheme (L1/L2 혼합형 중계 방법을 적용한 이더넷 기반 비압축 오디오 분배 시스템의 성능 분석)

  • Nam, Wie-Jung;Yoon, Chong-Ho;Park, Pu-Sik;Jo, Nam-Hong
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.46 no.12
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    • pp.108-116
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    • 2009
  • In this paper, we propose a Ethernet based audio distribution system with a new L1/L2 hybrid switching scheme, and evaluate its performance. The proposed scheme not only offers guaranteed low latency and jitter characteristics that are essentially required for the distribution of high-quality uncompressed audio traffic, and but also provide an efficient transmission of data traffic on the Ethernet environment. The audio distribution system with a proposed scheme consists of a master node and a number of relay nodes, and all nodes are mutually connected as a daisy-chain topology through up and downlinks. The master node generates an audio frame for each cycle of 125us, and the audio frame has 24 time slotted audio channels for carrying stereo 24 channels of 16-bit PCM sampled audio. On receiving the audio frame from its upstream node via the downlink, each intermediate node inserts its audio traffic to the reserved time slot for itself, then relays again to next node through its physical layer(L1) transmission - repeating. After reaching the end node, the audio frame is loopbacked through the uplink. On repeating through the uplink, each node makes a copy of audio slot that node has to receive, then play the audio. When the audio transmission is completed, each node works as a normal L2 switch, thus data frames are switched during the remaining period. For supporting this L1/L2 hybrid switching capability, we insert a glue logic for parsing and multiplexing audio and data frames at MII(Media Independent Interlace) between the physical and data link layers. The proposed scheme can provide a good delay performance and transmission efficiency than legacy Ethernet based audio distribution systems. For verifying the feasibility of the proposed L1/L2 hybrid switching scheme, we use OMNeT++ as a simulation tool with various parameters. From the simulation results, one can find that the proposed scheme can provides outstanding characteristics in terms of both jitter characteristic for audio traffic and transmission efficiency of data traffics.

The International Arbitration System for the Settlement of Investor-State Disputes in the FTA (FTA(자유무역협정)에서 투자자 대 국가간 분쟁해결을 위한 국제중재제도)

  • Lee, Kang-Bin
    • THE INTERNATIONAL COMMERCE & LAW REVIEW
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    • v.38
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    • pp.181-226
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    • 2008
  • The purpose of this paper is to describe the settling procedures of the investor-state disputes in the FTA Investment Chapter, and to research on the international arbitration system for the settlement of the investor-state disputes under the ICSID Convention and UNCITRAL Arbitration Rules. The UNCTAD reports that the cumulative number of arbitration cases for the investor-state dispute settlement is 290 cases by March 2008. 182 cases of them have been brought before the ICSID, and 80 cases of them have been submitted under the UNCITRAL Arbitration Rules. The ICSID reports that the cumulative 263 cases of investor-state dispute settlement have been brought before the ICSID by March 2008. 136 cases of them have been concluded, but 127 cases of them have been pending up to now. The Chapter 11 Section B of the Korea-U.S. FTA provides for the Investor_State Dispute Settlement. Under the provisions of Section B, the claimant may submit to arbitration a claim that the respondent has breached and obligation under Section A, an investment authorization or an investment agreement and that the claimant has incurred loss or damage by reason of that breach. Provided that six months have elapsed since the events giving rise to the claim, a claimant may submit a claim referred to under the ICSID Convention and the ICSID Rules of Procedure for Arbitration Proceedings; under the ICSID Additional Facility Rules; or under the UNCITRAL Arbitration Rules. The ICSID Convention provides for the jurisdiction of the ICSID(Chapter 2), arbitration(Chapter 3), and replacement and disqualification of arbitrators(Chapter 5) as follows. The jurisdiction of the ICSID shall extend to any legal dispute arising directly out of an investment, between a Contracting State and a national of another Contracting State, which the parties to the dispute consent in writing to submit to the ICSID. Any Contracting State or any national of a Contracting State wishing to institute arbitration proceedings shall address a request to that effect in writing to the Secretary General who shall send a copy of the request to the other party. The tribunal shall consist of a sole arbitrator or any uneven number of arbitrators appointed as the parties shall agree. The tribunal shall be the judge of its own competence. The tribunal shall decide a dispute in accordance with such rules of law as may be agreed by the parties. Any arbitration proceeding shall be conducted in accordance with the provisions of the Convention Section 3 and in accordance with the Arbitration Rules in effect on the date on which the parties consented to arbitration. The award of the tribunal shall be in writing and shall be signed by members of the tribunal who voted for it. The award shall deal with every question submitted to the tribunal, and shall state the reason upon which it is based. Either party may request annulment of the award by an application in writing addressed to the Secretary General on one or more of the grounds under Article 52 of the ICSID Convention. The award shall be binding on the parties and shall not be subject to any appeal or to any other remedy except those provided for in this Convention. Each Contracting State shall recognize an award rendered pursuant to this convention as binding and enforce the pecuniary obligations imposed by that award within its territories as if it were a final judgment of a court in that State. In conclusion, there may be some issues on the international arbitration for the settlement of the investor-state disputes: for example, abuse of litigation, lack of an appeals process, and problem of transparency. Therefore, there have been active discussions to address such issues by the ICSID and UNCITRAL up to now.

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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Isolation and Characterization of a Novel Flavonoid 3'-Hydroxylase (F3'H) Gene from a Chrysanthemum (Dendranthema grandiflorum) and Its Gamma-ray Irradiated Mutants (감마선 처리에 의한 스프레이형 국화 화색변이체로부터 Flavonoid 3'-Hydroxylase(F3'H) 유전자의 분리 및 특성 구명)

  • Chung, Sung-Jin;Lee, Geung-Joo;Kim, Jin-Baek;Kim, Dong-Sub;Kim, Sang-Hoon;Kang, Si-Yong
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.162-170
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    • 2012
  • The objectives of this study were to isolate and the sequence of novel $F3'H$ gene related to an anthocyanin pathway, and to confirm the expression patterns of the gene involved in the flower color variations of chrysanthemum mutants. In this study, we isolated the full-length cDNAs and the genomic DNAs of an $F3'H$ gene from a wild type (WT) chrysanthemum (cv. Argus) and its three color mutants. The sequence analysis revealed a putative open reading frame of 1,527 bp that encodes a polypeptide of 509 amino acids. Sequence homology ranged from 97% to 99% between 'Argus' and its three color mutants. The sequence analysis from the genomic DNA revealed that the chrysanthemum $DgF3'H$ gene consisted of three exons and two introns spanning a 3,830 bp length. The sizes of the gene for three mutants ranged from a shorter size of 3,828 bp to a longer size of 3,838 bp when compared to the size of WT. The total size of the two introns was 2,157 bp for WT, but those for three color mutants ranged from 2,154 bp to 2,159 bp. A result of an RT-PCR analysis indicated that the color variations of the mutants AM1 and AM2 can be partly explained by the structural modification derived from the sequencial changes in the gene caused by gamma ray. A Southern blot analysis revealed that the $DgF3'H$ gene existing as multiple copies in the chrysanthemum genome. A systemic study will be further needed to provide a genetic mechanism responsible for the color mutation and to uncover any involvement of genetic elements for the expression of the $DgF3'H$ gene for the color variation in chrysanthemum.

A Study on the Ijori Tortoise Pedestal of Namsan Mountain in Gyeong-Ju (경주 남산 이조리귀부에 관한 고찰)

  • Lee, Eun Seok;Cho, Hyun Kyung
    • Korean Journal of Heritage: History & Science
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    • v.43 no.4
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    • pp.56-77
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    • 2010
  • The tortoise pedestal for the memorial monument of Choe Jin-rip(an army officer in the mid-Joseon Period) in Ijo-ri, Naenam-myeon, Gyeongju is known to have been made in 1740. As such, it was originally understood to be a Joseon imitation of a tortoise pedestal made in the Unified Silla Period. The style of the Ijori Tortoise Pedestal differs from other tortoise pedestals dating back to the same period, and bears no resemblance to the Unified Silla pedestals of which it is a copy. Mullu ilgi, a record of the production of the pedestal, explains that the monument was made before the pedestal. Traces show that the two sides of the bottom of the monument were cut off so that it would fit into the smaller space made on the pedestal. It is scarcely conceivable that they made the pedestal and the platform without considering the bottom size of the monument. The record only states that the monument was made at a temple site named Baegundae, without explaining the details of the production process. This leaves some doubt as to whether its production was undertaken systematically. The cloud patterns engraved on this pedestal look similar to the temporal seriation found on the Tortoise Pedestal of the Royal Tomb of King Muyeol and the Seoangni Tortoise Pedestal of Gyeongju. The lotus pattern decorating the square pedestal on the back of the tortoise is one of a number of patterns that were widely used on roof-tiles in the 8th century, the heyday of the Unified Silla Kingdom. The Ijori Tortoise Pedestal, which represents a tortoise moving forward, displays a liveliness the like of which is rarely found in its cousins remaining in Gyeongju. The layout of the patterns in a queue on the tortoise-shell looks much better schematized than those made at an earlier date. It also looks like a more developed form, with the use of space taken into account. Such factors as the style of the patterns, the incongruity between the monument and its pedestal, and what is stated in the historical record indicate that the Ijori Tortoise Pedestal of Gyeongju was made in the mid-8th century(i.e. during the Unified Silla Period), rather than in the Joseon Period(i.e. the 18th century), as an imitation of earlier ones, including changes in the style unique to the Silla Period.