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      • 배추 시스테인 단백질 분해 효소 억제 유전자 BCPI-1의 발현과 종자 발아 및 유묘 생장과의 연관성

        홍준기,이은영,김정률,양경애,최영주,정우식,김호일,윤대진,이상열,조무제,임체오 Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-

        Phytocystatins are protein inhibitors of cysteine proteinases of the papain family that have been identified in both monocot and dicot plants. A cDNA encoding a phytocystatin, BCPI-1 (Brassica Cysteine Proteinase Inhibitor-1) has been isolated from Chinese cabbage (Brassica campestris L. ssp. pekinensis) flower buds. Here, we tested whether BCPI-1 transcription is regulated by hormones, and could be involved in regulating cysteine proteinases during and after germination. BCPI-1 was sensitive to exogenous GA_(3) and ABA, which are important factors controlling seed germination, suggesting that the expression of BCPI-1 is hormonally regulated. We introduced a recombinant plasmid containing the full-length BCPI-1 cDNA under the control of cauliflower mosaic virus 35S promoter into rice embryogenic calli using the particle delivery method, and regenerated a number of transgenic rice plants. Constitutively over-expressed BCPI-1 caused changes in overall plant growth and development, including reduced germination and seedling growth. These data support the role of the BCPI-1 in the regulation of endogenous proteinases during both seed germination and subsequent seedling development. Phylocystatins은 papain계열의 cysteine 단백질 분해 효소 활성을 특이적으로 억제하는 억제자로 다양한 식물 종으로부터 분리되었다. 본고에서는, 배추 화아 cDNA library로부터 분리된 phytocystatin인 BCPI-1(Brassica Cysteine Proteinase Inhibitor-1)의 생체 내 기능에 대해 연구하였다. 먼저, BCPI-1 전사체는 발아 조절에 중요한 영향을 미치는 GA₃와 ABA에 의해 예민하게 증가, 혹은 감소되는 반응을 보임으로써, BCPI-1이 식물 호르몬의 영향을 받으며, 특히 발아나 유묘의 생장 조절에 관련이 있음을 알 수 있었다. Particle bombardment 방법을 통하여 BCPI-1을 벼의 배 형성 세포 내에 도입, 재분화 시켜 형질 전환 벼를 생성하였다. CaMV 35S promoter의 조절에 의해 지속적인 BCPI-1 발현을 보이는 형질 전환 벼의 경우, 발아와 유묘 생장이 현저히 지연되었다. 위의 결과들을 바탕으로, BCPI-1이 식물체 내에서 생성되는 cysteine 계열의 단밸질 분해 효소 활성을 조절하여 종자의 발아와 유묘의 생장에 영향을 미치는 것으로 추측한다.

      • Gilbert 증후군에서 열량 제한 시험과 Phenobarbital 자극 시험의 의의(14예)

        이헌영,채경훈,정재훈,강윤세,김연수,문희석,박기오,이엄석,김선문,김석현,성재규,이병석,이강욱 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2

        Gilbert 증후군은 인구의 7%에서까지 나타날 수 있는 매우 흔한 증후군으로서 비진행성인 양성의 만성적 경과를 치하며, 간질환의 증상과 징후가 없는 경한 비포합형 고빌리루빈혈증이 특징인 일종의 체질적인 증상으로서 혈장 빌리루빈 농도에 대한 사춘기의 영향 때문에 10대와 20대에 자주 진단이 된다. 따라서 임상적인 중요성은 미약하지만 높은 빈도가 예상되는 점에 그 중요성이 부여되어야 할 것이다. 따라서 적정한 임상적 진단법으로 기왕에 소개된 열량제한 시험과 phenobarbital 유도 시험을 시행하고 이들의 진단적 가치를 알아보기 위하여 본 연구를 시행하였다. 1990년 7월부터 1999년 4월까지 충남대학교병원에 내원하여 HBsAg, IgG anti-HBc 및 anti-HCV가 음성이고, 간 초음파 스캔에서 이상이 없으며, 혈청 AST, ALT 및 AP가 정상인 비음주자에서 경한 비포합형 고빌리루빈혈증이 있는 14예의 환자들을 대상으로 ^(99m)Tc-DISID 스캔을 시행하였으며, 기저 치 총빌리루빈 및 포합형 빌리루빈 치를 측정한 다음에 하루에 400Kcal로 48시간동안 제한한 열량 제한 시험을 시행하였고, phenobarbital을 하루 60mg씩 5일간 투여한 후에도 각각 총빌리루빈과 포합형 빌리루빈 치를 검사하여 비포합형을 구하였다. 대상 환자들은 모두 14예로서 남자가 11예(78.6%)였고 여자가 3예(21.4%)여서 3.7:1로 남자에서 많았으며, 20대가 6예(42.9%), 30대가 역시 6예(42.9%) 및 40대가 2예(14.2%)로서 2,30대가 대부분(85.8%)이었다. 열량 제한 시험 후의 총빌리루빈 치, 비포합형 및 포합형 빌리루빈 치들은 평균 각각 5.5±2.7, 4.2±2.3 및 1.3±10mg/dL 로서, 시험 전 치들인 3.0±0.8, 2.2±0.8 및 0.7±0.4mg/dL 보다 유의하게(p=0.001, p=0.001, p=0.023) 상승하였다. 포합형 빌리루빈 치도 유의하게 상승하였으나 비포합형의 상승보다는 훨씬 낮아서 주로 비포합형이 증가하였다. phenobarbital 투여 중 설사가 발생하여 중단한 1예를 제외한 13예에서 열량 제한 시험 후에 상승하였던 총, 비포합형 및 포합형 빌리루빈 치가 phenobarbital 유도 시험후에는 2.0±1.1, 1.5±0.8 및 0.5±0.4mg/dL로서 열량 제한 시험 결과보다 유의하게 낮아졌고(p=0.00, p=0.000, p=0.001), 열량 제한 시험 전의 기초치들인 3.0±0.8, 2.2±0.8 및 0.7±0.4mg.dL 보다도 더욱 낮아졌으며 유의한 차이(p=0.001, p=0.02, p=0.005)를 나타내었다. 14예에서 시행한 ^(99m)-Tc DISIDA 스캔에서 9예(64.3%)가 정상이었고, 5예(35.7%)에서는 심장 및 신장으로의 간외 섭취가 3예였고, 60분까지 소장 배출이 없는 배설 지연 예와 담낭 수축 불량 예가 각각 1예 씩 발견되었다. Phenobarbital 투여시험에서 민감도가 열량제한시험에 비해 더 높았다(92.3%와 50.0%). Gilbert 증후군에서 1일 400 Kcal로 48시간의 열량제한 시험과 1일 60mg의 phenobarbital을 5일간 투여하는 유도 시험은 편리하고 유용한 임상적인 진단법으로 이용할 수 있다고 생각된다. 그러나 열량 제한 시험에서는 증가 기준의 통일이 필요하다고 유추되며 phenobarbital 유도 시험이 민감도가 더 높은 것으로 생각된다. Gilbert's syndrome is very frequent and benign chronic process characterized by mild, intermittent, unconjugated hyperbilirubinemia without any symptom and sign of liver disease. Previously intoduced caloric restriction test and phenobarbital stimulation test as two appropriate clinical tests had been examined and their diagnostic values were reevaluated. Fourteen patients with mild, persistent, unconjugated hyperbilirubinemia were included. Subsequently caloric restriction has been applicated by 400 Kcal/day for 48 hours and phenobarbital has been prescribed by 60 mg/day for 5 days. Therafter serum levels of total and direct bilirubin were measured. Most of the patients were third and fourth decade(85.8%) and male predominant. Each basal serum levels of total, indirect and direct bilirubin were 3.0±0.8, 2.2±0.8 and 0.7±0.4 mg/dL. After caloric restriction test, each levels were increased significantly to 5.5±2.7, 4.2±2.3 and 1.3±1.0 mg/dL(p=0.001, p=0.001, p=0.023). After phenobarbital stimulation test for 13 patients had been practiced, increased levels of each bilirubin after caloric restriction test were decreased significantly to 2.0±1.0, 1.5±0.8 and 0.5±0.4 mg/dL(p=0.000, p=0.000, p=0.001) and these levels were significantly lower than basal levels(p=0.001, p=0.02, p=0.005). The sensitivities of caloric restriction test were 85.7%, 50.0%, and 71.4%, 35.7%(1.0, 1.5 mg increase of total bilirubin and 1.0, 1.5 mg/dL increase of indirect bilirubin). The sensitivities of phenobarbital stimulation test were 93.2% and 92.3% at criteria of 1.5 mg/dL increase of total bilirubin and indirect bilirubin. On the diagnosis of Gilbert syndrome, caloric restriction test and phenobarbital stimulation test are convenient and useful diagnostic tools in clinical face. And also phenobarbital stimulation test has higher sensitivity than caloric restriction test. Furthermore, standardization of bilirubin increment would be necessary in caloric restriction test.

      • SCISCIESCOPUS

        Interleukin-6 and Cyclic AMP Stimulate Release of Cathepsin B in Human Osteoblasts

        Chae, Han-Jung,Ha, Ki-Chan,Lee, Geun-Youn,Yang, Sun-Kyung,Yun, Ki-Jung,Kim, Eun-Cheol,Kim, Sun-Hee,Chae, Soo-Wan,Kim, Hyung-Ryong Taylor & Francis 2007 IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY Vol.29 No.2

        <P> Previous studies have suggested that cathepsin B participates in the joint destruction associated with rheumatoid arthritis (RA). This study examined the activity of cathepsin B (a lysosomal cysteine protease) in human osteoblasts along with its regulation by cyclic AMP and Interleukin-6 (IL-6). Cyclic AMP elevating agents activate cathepsin B and stimulate the secretion of cathepsin B via the secretion of IL-6, a potent mediator of RA. This study investigated the induction of cathepsin B using the proinflammatory cytokine in human osteoblasts (MG-63) in relation to p38 mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-kappa B transcription factor. When added to MG-63 cells, IL-6 stimulated the production of cathepsin B, which was reduced significantly by the addition of SB203580, a specific p38 MAPK inhibitor. In addition, the release of IL-6 was also inhibited by either pyrrolidine dithiocarbamate (PDTC) or NF-&kgr;B SN50, which are potent NF-&kgr;B inhibitors. Both NF-&kgr;B inhibitors had a larger inhibitory effect on the activity of cathepsin B in the presence of SB203580. IL-6 stimulated the NF-&kgr;B binding affinity as well as the activation of p38 MAP kinase, leading to the release of cathepsin B. However, SB203580 had no effect on the IL-6-induced activation of NF-&kgr;B, and neither of the NF-&kgr;B inhibitors decreased the level of p38 MAPK activation in the IL-6-stimulated osteoblasts. Moreover, IL-6 increased the activity of urokinase type plasminogen activator (uPA) in MG-63 cells, which was inhibited by SB203580, PDTC and NF-&kgr;B SN50. This strongly suggests that p38 MAPK and NF-&kgr;B are essential to the IL-6-induced activation of cathepsin B or uPA and that these two IL-6-activated pathways can act independently.</P>

      • SCISCIESCOPUS

        Characterization of <i>Salmonella enterica</i> serovar Typhimurium and <i>Salmonella enterica</i> serovar 4,[5],12:i:- isolates from pigs presenting with diarrhea in Korea

        LEE, Ki-Eun,LEE, Deog-Yong,CHOI, Hwan-Won,CHAE, Su-Jin,YUN, Young-Sun,LEE, Ki-Chan,CHO, Yun-Sang,YANG, Dong-Kun The Japanese Society of Veterinary Science 2015 The Journal of veterinary medical science Vol.77 No.11

        <P>Between 2011 and 2012, a total of 896 pig fecal samples were collected from nine provinces in Korea, and 50 <I>salmonella enterica </I>susp. <I>enterica</I> serovar Typhimurium (<I>S</I>. Typhimurium) was isolated. The characteristics of the 50 strains were analyzed, and 4 strains were identified as <I>Salmonella enterica</I> subsp. <I>enterica</I> serovar 4,[5],12:i:-. <I>Salmonella</I> 4,[5],12:i:- could not be distinguished from <I>S</I>. Typhimurium through phage typing, antimicrobial resistance testing or multiple-locus variable-number tandem repeat analysis (MLVA). However, among the four <I>Salmonella</I> 4,[5],12:i:- strains, one (KVCC-BA1400078) was identified as a <I>Salmonella</I> 4,[5],12:i:- clone isolated from humans in the United States, and another (KVCC-BA1400080) was identified as DT193, which has been primarily isolated from humans and animals in European countries. The presence of <I>Salmonella</I> 4,[5],12:i:- in Korea poses a significant threat of horizontal transfer between pigs and humans.</P>

      • KCI등재

        Identification of Single Nucleotide Polymorphisms in the TNFRSF17 Gene and Their Association with Gastrointestinal Disorders

        Chae, Soo-Cheon,Yu, Ji-In,Oh, Gyung-Jae,Choi, Chang-Soo,Choi, Suck-Chei,Yang, Yun-Sik,Yun, Ki-Jung Korean Society for Molecular and Cellular Biology 2010 Molecules and cells Vol.29 No.1

        TNFRSF17 is preferentially expressed in mature B lymphocytes, and may be important for the development of B cells. TNFRSF17 is selected as a candidate susceptibility gene to IBD pathogenesis by our cDNA microarray analysis, and we showed the specific expression of TNFRSF17 in resting and activated $CD19^+$ cells obtained from human blood. We identified four SNPs (g-1729G>A, g.2295T>C, g.2445G>A and g.2493G>A) and one variation site (g.-894delT) in the TNFRSF17 gene using direct sequencing analysis. In addition, the association of the genotype and allelic frequencies of these SNPs was studied in healthy controls and in patients with ulcerative colitis (UC) or irritable bowel syndrome (IBS). Although, the genotype and allelic frequencies of these SNPs, in the UC and IBS patients, were not significantly different from those in the healthy controls, the distribution of the AAG, GGA, AGG and AAA haplotypes, of the SNPs (g.-1729G>A, g.2445G>A and g.2493G>A) associated with the TNFRSF17 gene, in the UC patients, were notably different from those of the healthy controls (P = 0.002, 0.002, 4.7E-4 and 3.3E-6, respectively). Moreover, the frequencies of the AAG, AGG, GAG and GAA haplotypes were significantly different in the IBS patients compared to the healthy controls (P = 4.2E-5, 4.4E-17, 1.8E-22 and 1.6E-10, respectively). These results suggest that the haplotypes of the TNFRSF17 polymorphisms might be associated with UC and IBS susceptibility.

      • 3D intra-stacked CoO/carbon nanocomposites welded by Ag nanoparticles for high-capacity, reversible lithium storage.

        Chae, Changju,Kim, Ki Woong,Kim, Sue Jin,Lee, Daehee,Jo, Yejin,Yun, Young Jun,Moon, Jooho,Choi, Youngmin,Lee, Sun Sook,Choi, Sungho,Jeong, Sunho RSC Pub 2015 Nanoscale Vol.7 No.23

        <P>A wet-chemical, facile strategy is proposed for forming three-dimensional intra-structured nanocomposites to facilitate the development of high performance anodes for lithium ion batteries. The nanocomposites are composed of cobalt oxide nanoparticles, reduced graphene oxides, and Ag nanoparticles, and all the constituent materials are incorporated homogenously in a layer-by-layer structured geometry by a simple sono-chemical hybridizing process in a single, one-pot batch. Herein, it is revealed that the homogenously intra-stacked oxide, carbon, and metallic phases play critical roles in determining electrochemical performance (i.e. high capacity, rate capability, and cycling stability) of nanocomposite-based anodes, owing to the characteristic chemical/physical nature of constituent materials welded by partial melting of the metallic nanoparticles. In particular, by virtue of a characteristic role of a nano-Ag phase in suppressing the irreversible capacity, a critical drawback for metal oxide-based anodes, excellent capacities are demonstrated (983 and 770 mA h g(-1) at current densities of 100 and 2000 mA g(-1), respectively).</P>

      • KCI등재

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