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      • An S-locus receptor-like kinase in plasma membrane interacts with calmodulin in <i>Arabidopsis</i>

        Kim, Ho Soo,Jung, Mi Soon,Lee, Kyunghee,Kim, Kyung Eun,Yoo, Jae Hyuk,Kim, Min Chul,Kim, Doh Hoon,Cho, Moo Je,Chung, Woo Sik Elsevier 2009 FEBS letters Vol.583 No.1

        <P><B>Abstract</B></P><P>Calmodulin-regulated protein phosphorylation plays a pivotal role in amplifying and diversifying the action of calcium ion. In this study, we identified a calmodulin-binding receptor-like protein kinase (CBRLK1) that was classified into an S-locus RLK family. The plasma membrane localization was determined by the localization of CBRLK1 tagged with a green fluorescence protein. Calmodulin bound specifically to a Ca<SUP>2+</SUP>-dependent calmodulin binding domain in the C-terminus of CBRLK1. The bacterially expressed CBRLK1 kinase domain could autophosphorylate and phosphorylates general kinase substrates, such as myelin basic proteins. The autophosphorylation sites of CBRLK1 were identified by mass spectrometric analysis of phosphopeptides.</P><P><B>Structured summary</B></P><P>MINT-6800947:<I>CBRLK1</I> (uniprotkb:Q9ZT06) and <I>AtCaM2</I> (uniprotkb:P25069) <I>bind</I> (MI:0407) by <I>electrophoretic mobility shift assay</I> (MI:0413)</P><P>MINT-6800966:<I>AtCaM2</I> (uniprotkb:P25069) and <I>CBRLK1</I> (uniprotkb:Q9ZT06) <I>bind</I> (MI:0407) by <I>competition binding</I> (MI:0405)</P><P>MINT-6800930:<I>CBRLK1</I> (uniprotkb:Q9ZT06) <I>binds</I> (MI:0407) to <I>AtCaM2</I> (uniprotkb:P25069) by <I>far Western blotting</I> (MI:0047)</P><P>MINT-6800978:<I>AtCaM2</I> (uniprotkb:P25069) <I>physically interacts</I> (MI:0218) with <I>CBRLK1</I> (uniprotkb:Q9ZT06) by <I>cytoplasmic complementation assay</I> (MI:0228)</P>

      • Histone deacetylase 3 is selectively involved in L3MBTL2-mediated transcriptional repression

        Yoo, Jung-Yoon,Choi, Kyung-Chul,Kang, HeeBum,Kim, Young Jun,Lee, Jeongmin,Jun, Woo Jin,Kim, Mi-Jeong,Lee, Yoo-Hyun,Lee, Ok-Hee,Yoon, Ho-Geun Elsevier 2010 FEBS letters Vol.584 No.11

        <P><B>Abstract</B></P><P>This is the first report that L(3)mbt-like 2 (L3MBTL2) specifically interacts with the histone deacetylase domain of histone deacetylase 3 (HDAC3) via its MBT domain. Here, we show that L3MBTL2 selectively interacts with HDAC3, but not other class I HDACs. An in vitro peptide-binding assay demonstrated the specific association of HDAC3 with methylated histone-K20 tail and L3MBTL2. Furthermore, depletion of HDAC3 resulted in a decrease of methylated K20-H4, as well as an increase in acetylated histone H3. Consequently, HDAC3 knock-down selectively suppressed L3MBTL2-mediated transcriptional repression. Taken together, our results reveal the concerted action of both HDAC3 and L3MBTL2 in histone deacetylation and methylation-dependent transcriptional repression.</P><P><B>Structured summary</B></P><P>MINT-7719975: <I>L3MBTL2</I> (uniprotkb:Q969R5) and <I>HDAC3</I> (uniprotkb:O15379) <I>colocalize</I> (MI:0403) by <I>fluorescence microscopy</I> (MI:0416)</P><P>MINT-7719941, MINT-7719921: <I>L3MBTL2</I> (uniprotkb:Q969R5) <I>binds</I> (MI:0407) to <I>HDAC3</I> (uniprotkb:O15379) by <I>pull down</I> (MI:0096)</P><P>MINT-7719991: <I>HDAC3</I> (uniprotkb:O15379) <I>physically interacts</I> (MI:0915) with <I>L3MBTL2</I> (uniprotkb:Q969R5) by <I>anti bait coimmunoprecipitation</I> (MI:0006)</P><P>MINT-7719958: <I>L3MBTL2</I> (uniprotkb:Q969R5) <I>physically interacts</I> (MI:0915) with <I>HDAC3</I> (uniprotkb:O15379) by <I>anti tag coimmunoprecipitation</I> (MI:0007)</P><P>MINT-7719897: <I>HDAC3</I> (uniprotkb:O15379) <I>physically interacts</I> (MI:0915) with <I>L3MBTL2</I> (uniprotkb:Q969R5) by <I>two hybrid</I> (MI:0018)</P>

      • KCI등재

        한국인 직무 스트레스 측정도구의 개발 및 표준화

        장세진,고상백,강동묵,김성아,강명근,이철갑,정진주,조정진,손미아,채창호,김정원,김정일,김형수,노상철,박재범,우종민,김수영,김정연,하미나,박정선,이경용,김형렬,공정옥,김인아,김정수,박준호,현숙정,손동국 大韓産業醫學會 2005 대한직업환경의학회지 Vol.17 No.4

        Background and Purposes: Over the past three decades, numerous studies performed in Korea have reported that job stress is a determinant risk factor for chronic diseases and work disability. Every society has its own culture and occupational climate particular to their organizations, and hence experiences different occupational stress. An occupational stress measurement tool therefore needs to be developed to estimate it objectively. The purpose of this study is to develop and standardize the Korean Occupational Stress Scale (KOSS) which is considered to be unique and specific occupational stressors in Korean employees. Subjects and Methods: Data were obtained from the National Study for Development and Standardization of Occupational Stress (NSDSOS Project: 2002-2004). A total of 12,631 employees from a nationwide sample proportional to the Korean Standard Industrial Classification and the Korean Standard Occupational Classification were administered. The KOSS was developed for 2 years (2002-2004). In the first year, we collected 255 items from the most popular job stress measurement tools such as JCQ, ERI, NIOSH and OSI, and 44 items derived from the a qualitative study (depth interview). Forty-three items of KOSS, in the second year, were retained for use in the final version of the KOSS by using Delphi and factor analysis. Items were scored using conventional 1-2-3-4 Likert scores for the response categories. Results: We developed eight subscales by using factor analysis and validation process: physical environment (3 items), job demand (8 items), insufficient job control (5 items), interpersonal conflict (4 items), job insecurity (6 items), organizational system (7 items), lack of reward (6 items), and occupational climate (4 items). Together they explained 50.0% of total variance. Internal consistency alpha scores were ranged from 0.51 to 0.82. Twenty-four items of the short form of the KOSS (KOSS-SF) were also developed to estimate job stress in the work setting. Because the levels of the subscales of occupational stress were gender dependent, gender-specific standard norms for both the 43-item full version and the 24-item short form using a quartile for the subscales of KOSS were presented. Conclusion: The results of this study suggest that KOSS might be an appropriate measurement scale to estimate occupational stress of Korean employees. Further and more detailed study needs to be conducted to improve the validity of this scale.

      • SCOPUSKCI등재

        Function of the pentose phosphate pathway and its key enzyme, transketolase, in the regulation of the meiotic cell cycle in oocytes

        Kim, Yunna,Kim, Eun-Young,Seo, You-Mi,Yoon, Tae Ki,Lee, Woo-Sik,Lee, Kyung-Ah The Korean Society for Reproductive Medicine 2012 Clinical and Experimental Reproductive Medicine Vol.39 No.2

        Objective: Previously, we identified that transketolase (Tkt), an important enzyme in the pentose phosphate pathway, is highly expressed at 2 hours of spontaneous maturation in oocytes. Therefore, this study was performed to determine the function of Tkt in meiotic cell cycle regulation, especially at the point of germinal vesicle breakdown (GVBD). Methods: We evaluated the loss-of-function of Tkt by microinjecting Tkt double-stranded RNAs (dsRNAs) into germinal vesicle-stage oocytes, and the oocytes were cultured in vitro to evaluate phenotypic changes during oocyte maturation. In addition to maturation rates, meiotic spindle and chromosome rearrangements, and changes in expression of other enzymes in the pentose phosphate pathway were determined after Tkt RNA interference (RNAi). Results: Despite the complete and specific knockdown of Tkt expression, GVBD occurred and meiosis was arrested at the metaphase I (MI) stage. The arrested oocytes exhibited spindle loss, chromosomal aggregation, and declined maturation promoting factor and mitogen-activated protein kinase activities. The modified expression of two enzymes in the pentose phosphate pathway, Prps1 and Rbks, after Tkt RNAi and decreased maturation rates were amended when ribose-5-phosphate was supplemented in the culture medium, suggesting that the Tkt and pentose phosphate pathway are important for the maturation process. Conclusion: We concluded that Tkt and its associated pentose phosphate pathway play an important role in the MI-MII transition of the oocytes' meiotic cell cycle, but not in the process of GVBD.

      • KCI등재

        중환자실 전문간호사의 전문간호행위 분류와 수행분석

        김진현,김명애,김미원,김경숙,유정숙,이은희 대한간호학회 간호행정학회 2009 간호행정학회지 Vol.15 No.4

        Purpose: The purpose of this study was to reclassify the advanced nursing practices of critical care nurse practitioners(CCNPs) in intensive care unit and measure the time and frequency of CCNP's activities. Method: Practices of ICU nurses are divided into RN's and CCNP's practices by a panel of ICU nursing experts. Each practice of CCNP is defined and CCNP's working time and service frequencies are monitored in general hospitals. Result: Practices of CCNP were classified into 4 domains and 32 practices. Fourteen practices by CCNPs were completed in 10 minutes and the other 12 practices consumed 10-30 minutes. A priority of practice in respiratory therapy was given to artificial airway management, management of tracheostomy patient, lower respiratory care, and the priority of CRRT was management of anticoagulation. Conclusions: Advanced nursing practices of CCNPs were recognized from those of RNs. A further research of CCNPs practices should be extended to other advanced practices and it is required to evaluate economic value of advanced nursing practice in the national health insurance system.

      • KCI등재

        단클론항체를 이용한 타액 내 Streptococcus mutans 수준의 측정

        김추성,김재곤,양연미,백병주,이경열,김미아,임수민 大韓小兒齒科學會 2010 大韓小兒齒科學會誌 Vol.37 No.2

        Streptococcus mutans는 구강 내에 상존하는 치아우식증의 주요 원인균으로서 치면의 피막에 부착 후 glucan을 형성하여 세균의 군락을 이루며, 외부로부터 공급된 자당대사를 통하여 유기산을 생성함으로써 법랑질을 탈회시킨다. 치아우식 활성도의 평가를 위한 단클론항체를 이용한 방법은 진료실에서 빠른 시간 내에 간편하게 타액에 존재하는 Streptococcus mutans의 정량분석이 가능한 방법이다. 이 연구는 3세에서 6세 사이의 어린이 15명을 대상으로 자극성 타액을 채취하여 시판 중인 단클론항체를 이용한 Salivacheck Mutans, strip을 이용한 Dentocult-SM 그리고 MSB배지 배양법으로서 타액 내 Streptococcus mutans를 측정한 후 그 값을 우식경험치아수와 비교하여 상관관계를 알아보았다. Saliva-check Mutans를 이용한 방법은 Dentocult-SM과 MSB배지법과 통계학적으로 유의한 상관관계를 보였으나 (p<0.05), MSB배지법은 어린이의 우식경험치아수와 통계학적으로 유의한 결과를 나타내지 않았다 (p=0.34). Streptococcus mutans, one of the major causal agents of dental caries, is component of the dental plaque. It produces various organic acids such as lactic acid which is the end-product of glycolysis, and this leads to dental caries. A new system using species-specific monoclonal antibodies was developed to detect Streptococcus mutans in saliva. The system quickly detects salivary Streptococcus mutans in 30min and classifies the result into two levels. The purpose of this study was to investigate correlation between monoclonal antibody-based detecting system and selective medium-based detecting methods. Children's deft indices were also compared with Streptococcus mutans counts in MSB agar plate. Subjects consisted of 15 children in the age of 3 to 6 years. They were assigned to three groups: Group Ⅰ (deft index = 3), Group Ⅱ (deft index ≤ 3), Group Ⅲ (deft index ≥ 4). The results are as follows : 1. The rate of children with positive response was 13.3% and with negative response was 86.7% in the result of Saliva-check Mutans test kit. 2. There was a positive correlation between monoclonal antibody-based detecting system and selective medium-based detecting methods (p<0.05). 3. Streptococcus mutans counts in MSB agar plate were irrelevant to deft of children(p=0.34).

      • KCI등재후보

        Identification of Genes Modulated by High Extracellular Calcium in Coculture of Mouse Osteoblasts and Bone Marrow Cells by Oligo Chip Assay

        Kim, Hyung-Keun,Song, Mina,Jun, ji-Hae,Woo, Kyung-Mi,Kim, Gwan-Shik,Baek, Jeong-Hwa The Korean Academy of Oral Biology 2006 International Journal of Oral Biology Vol.31 No.2

        Calcium concentration in the bone resorption lacunae is high and is in the mM concentration range. Both osteoblast and osteoclast have calcium sensing receptor in the cell surface, suggesting the regulatory role of high extracellular calcium in bone merabolism. In vitro, high extracellular calcium stimulated osteoclastogenesis in coculture of mouse osteoblasts and bone marrow cells. Therefore we examined the genes that were commonly regulated by both high extracellular calcium and 1,25(OH)_(2)vitaminD_(3)(VD3) by using mouse oligo 11 K gene chip. In the presence of 10 mM[Ca^(2+)]e or 10 nM VD3, mouse calvarial osteoblasts and bone marrow cells were co-cultured for 4 days when tartrate resistant acid phosphatase-positive multinucleated cells start to appear. Of 11,000 genes examined, the genes commonly regulated both by high extracellular calcium and by VD3 were as follows; 1) the expressions of genes which were osteoclast differentiation markers or were associated with osteoclastogenesis were up-regulated both by high extracellular calcium and by VD3; trap, mmp9, car2, ctsk, ckb, atp6b2, tm7sf4, rab7, 2) several chemokine and chemokine receptor genes such as sdf1, scya2, scyb5, scya6, scya8, scya9, and ccr1 were up-regulated both by high ectracellular calcium and by VD3, 3) the genes such as mmp1b, mmp3 and c3 which possibly stimulate bone resorption by osteoclast, were commonly up-regulated, 4) the gene such as c1q and msr2 which were related with macrophage function, were commonly down-regulated, 5) the genes which possibly stimulate osteoblast differentiation and/or mineralization of extracellular matrix, were commonly down-regulated;slc8a1, admr, plod2, lox, fosb, 6) the genes which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were commonly up-regulated;s100a4, npr3, mme, 7) the genes such as calponin 1 and tgfbi which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were up-regulated by high extracelluar calcium but were down-regulated by VD3. These results suggest that in coculture condition, both high extracellular calcium and VD3 commonly induce osteoclastogenesis but suppress osteoblast differentiation/mineralization by regulating the expression of related genes.

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