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      • KCI등재

        Economic Evaluation of Transmission Expansion for Investment Incentives in a Competitive Electricity Market

        Robert Fischer,Sung-Kwan Joo 대한전기학회 2008 International Journal of Control, Automation, and Vol.6 No.5

        With the shift of the electric power industry from a regulated monopoly structure to a competitive market environment, the focus of the transmission expansion planning has been moving from reliability-driven transmission expansion to market-based transmission expansion. In market-based transmission expansion, however, a growing demand for electricity, an increasing number of transmission bottlenecks, and the falling levels of transmission investment have created the need for an incentive to motivate investors. The expectation of profit serves as a motivational factor for market participants to invest in transmission expansion in a competitive market. To promote investment in transmission expansion, there is an increasing need for a systematic method to examine transmission expansion for investment incentives from multiple perspectives. In this paper, the transmission expansion problem in a competitive market environment is formulated from ISO and investors' perspectives. The proposed method uses parametric analysis to analyze benefits for investors to identify the most profitable location and amount for transmission addition. Numerical results are presented to demonstrate the effectiveness of the proposed method.

      • SCIESCOPUSKCI등재

        Microbiological cleaning and disinfection efficacy of a three-stage ultrasonic processing protocol for CAD-CAM implant abutments

        Gehrke, Peter,Riebe, Oliver,Fischer, Carsten,Weinhold, Octavio,Dhom, Gunter,Sader, Robert,Weigl, Paul The Korean Academy of Prosthodonitics 2022 The Journal of Advanced Prosthodontics Vol.14 No.5

        PURPOSE. Computer-aided design and manufacturing (CAD-CAM) of implant abutments has been shown to result in surface contamination from site-specific milling and fabrication processes. If not removed, these contaminants can have a potentially adverse effect and may trigger inflammatory responses of the peri-implant tissues. The aim of the present study was to evaluate the bacterial disinfection and cleaning efficacy of ultrasonic reprocessing in approved disinfectants to reduce the microbial load of CAD-CAM abutments. MATERIALS AND METHODS. Four different types of custom implant abutments (total N = 32) with eight specimens in each test group (type I to IV) were CAD-CAM manufactured. In two separate contamination experiments, specimens were contaminated with heparinized sheep blood alone and with heparinized sheep blood and the test bacterium Enterococcus faecium. Abutments in the test group were processed according to a three-stage ultrasonic protocol and assessed qualitatively and quantitatively by determination of residual protein. Ultrasonicated specimens contaminated with sheep blood and E. faecium were additionally eluted and the dilutions were incubated on agar plates for seven days. The determined bacterial counts were expressed as colony-forming units (CFU). RESULTS. Ultrasonic reprocessing resulted in a substantial decrease in residual bacterial protein to less than 80 ㎍ and a reduction in microbiota of more than 7 log levels of CFU for all abutment types, exceeding the effect required for disinfection. CONCLUSION. A three-stage ultrasonic cleaning and disinfection protocol results in effective bacterial decontamination. The procedure is reproducible and complies with the standardized reprocessing and disinfection specifications for one- or two-piece CAD-CAM implant abutments.

      • 5-Methylcytosine Recognition by <i>Arabidopsis thaliana</i> DNA Glycosylases DEMETER and DML3

        Brooks, Sonja C.,Fischer, Robert L.,Huh, Jin Hoe,Eichman, Brandt F. American Chemical Society 2014 Biochemistry Vol.53 No.15

        <P/><P>Methylation of cytosine to 5-methylcytosine (5mC) is important for gene expression, gene imprinting, X-chromosome inactivation, and transposon silencing. Active demethylation in animals is believed to proceed by DNA glycosylase removal of deaminated or oxidized 5mC. In plants, 5mC is removed from the genome directly by the DEMETER (DME) family of DNA glycosylases. <I>Arabidopsis thaliana</I> DME excises 5mC to activate expression of maternally imprinted genes. Although the related Repressor of Silencing 1 (ROS1) enzyme has been characterized, the molecular basis for 5mC recognition by DME has not been investigated. Here, we present a structure–function analysis of DME and the related DME-like 3 (DML3) glycosylases for 5mC and its oxidized derivatives. Relative to 5mC, DME and DML3 exhibited robust activity toward 5-hydroxymethylcytosine, limited activity for 5-carboxylcytosine, and no activity for 5-formylcytosine. We used homology modeling and mutational analysis of base excision and DNA binding to identify residues important for recognition of 5mC within the context of DNA and inside the enzyme active site. Our results indicate that the 5mC binding pocket is composed of residues from discrete domains and is responsible for discrimination against 5mC derivatives, and suggest that DME, ROS1, and DML3 utilize subtly different mechanisms to probe the DNA duplex for cytosine modifications.</P>

      • KCI등재

        Temporal and Spatial Downregulation of Arabidopsis MET1 Activity Results in Global DNA Hypomethylation and Developmental Defects

        김민희,Hyonhwa Ohr,Jee Woong Lee,현유봉,Robert L. Fischer,최연희 한국분자세포생물학회 2008 Molecules and cells Vol.26 No.6

        DNA methylation is an epigenetic mechanism for gene silencing. In Arabidopsis, MET1 is the primary DNA me-thyltransferase that maintains CG DNA methylation. Plants having an overall reduction of MET1 activity, caused by a met1 mutation or a constitutively expressed MET1 antisense gene, display genome hypomethylation, inappropriate gene and transposon transcription, and developmental abnormalities. However, the effect of a transient reduction in MET1 activity caused by inhibiting MET1 expression in a restricted set of cells is not known. For this reason, we generated transgenic plants with a MET1 antisense gene fused to the DEMETER (DME) promoter (DME:MET1 a/s). Here we show that DME is expressed in leaf primordia, lateral root primoridia, in the region distal to the primary root apical meristem, which are regions that include proliferating cells. Endogenous MET1 expression was normal in organs where the DME:MET1 a/s was not expressed. Although DME promoter is active only in a small set of cells, these plants displayed global developmental abnormalities. Moreover, centromeric repeats were hypomethylated. The developmental defects were accumulated by the generations. Thus, not maintaining CG methylation in a small population of proliferating cells flanking the meristems causes global developmental and epigenetic abnormalities that cannot be rescued by restoring MET1 activity. These results suggest that during plant development there is little or no short-term molecular memory for reestablishing certain patterns of CG methylation that are maintained by MET1. Thus, continuous MET1 activity in dividing cells is essential for proper patterns of CG DNA methylation and development.

      • An E3 ligase complex regulates SET-domain polycomb group protein activity in Arabidopsis thaliana.

        Jeong, Cheol Woong,Roh, Hyungmin,Dang, Tuong Vi,Choi, Yang Do,Fischer, Robert L,Lee, Jong Seob,Choi, Yeonhee National Academy of Sciences 2011 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.108 No.19

        <P>Transcriptional repression via methylation of histone H3 lysine 27 (H3K27) by the polycomb repressive complex 2 (PRC2) is conserved in higher eukaryotes. The Arabidopsis PRC2 controls homeotic gene expression, flowering time, and gene imprinting. Although downstream target genes and the regulatory mechanism of PRC2 are well understood, much less is known about the significance of posttranslational regulation of PRC2 protein activity. Here, we show the posttranslational regulation of CURLY LEAF (CLF) SET-domain polycomb group (PcG) protein by the F-box protein, UPWARD CURLY LEAF1 (UCL1). Overexpression of UCL1 generates mutant phenotypes similar to those observed in plants with a loss-of-function mutation in the CLF gene. Leaf curling and early flowering phenotypes of UCL1 overexpression mutants, like clf mutants, are rescued by mutations in the AGAMOUS and FLOWERING LOCUS T genes, which is consistent with UCL1 and CLF functioning in the same genetic pathway. Overexpression of UCL1 reduces the level of CLF protein and alters expression and H3K27 methylation of CLF-target genes in transgenic plants, suggesting that UCL1 negatively regulates CLF. Interaction of UCL1 with CLF was detected in plant nuclei and in the yeast two-hybrid system. The UCL1 F-box binds in vivo to components of the E3 ligase complex, which ubiquitylate proteins that are subsequently degraded via the ubiquitin-26S proteasome pathway. Taken together, these results demonstrate the posttranslational regulation of the CLF SET-domain PcG activity by the UCL1 F-box protein in the E3 ligase complex.</P>

      • KCI등재SCISCIE

        Temporal and spatial downregulation of Arabidopsis MET1 activity results in global DNA hypomethylation and developmental defects.

        Kim, Minhee,Ohr, Hyonhwa,Lee, Jee Woong,Hyun, Youbong,Fischer, Robert L,Choi, Yeonhee Korean Society for Molecular Biology 2008 Molecules and cells Vol.26 No.6

        <P>DNA methylation is an epigenetic mechanism for gene silencing. In Arabidopsis, MET1 is the primary DNA methyltransferase that maintains CG DNA methylation. Plants having an overall reduction of MET1 activity, caused by a met1 mutation or a constitutively expressed MET1 antisense gene, display genome hypomethylation, inappropriate gene and transposon transcription, and developmental abnormalities. However, the effect of atransient reduction in MET1 activity caused by inhibiting MET1 expression in a restricted set of cells is not known. For this reason, we generated transgenic plants with a MET1 antisense gene fused to the DEMETER (DME) promoter (DME:MET1 a/s). Here we show that DME is expressed in leaf primordia, lateral root primoridia, in the region distal to the primary root apical meristem, which are regions that include proliferating cells. Endogenous MET1 expression was normal in organs where the DME:MET1 a/s was not expressed. Although DME promoter is active only in a small set of cells, these plants displayed global developmental abnormalities. Moreover, centromeric repeats were hypomethylated. The developmental defects were accumulated by the generations. Thus, not maintaining CG methylation in a small population of proliferating cells flanking the meristems causes global developmental and epigenetic abnormalities that cannot be rescued by restoring MET1 activity. These results suggest that during plant development there is little or no short-term molecular memory for reestablishing certain patterns of CG methylation that are maintained by MET1. Thus, continuous MET1 activity in dividing cells is essential for proper patterns of CG DNA methylation and development.</P>

      • KCI등재

        Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

        Yeonhee Choi,Kyunghyuk Park,Jennifer M. Frost,Adam James Adair,Dong Min Kim,Hyein Yun,Janie S. Brooks,Robert L. Fischer 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.10

        The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dis-section followed by the derivation of central cell proto-plasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

      • SCISCIESCOPUS

        DNA demethylation is initiated in the central cells of <i>Arabidopsis</i> and rice

        Park, Kyunghyuk,Kim, M. Yvonne,Vickers, Martin,Park, Jin-Sup,Hyun, Youbong,Okamoto, Takashi,Zilberman, Daniel,Fischer, Robert L.,Feng, Xiaoqi,Choi, Yeonhee,Scholten, Stefan National Academy of Sciences 2016 Proceedings of the National Academy of Sciences Vol.113 No.52

        <P>Cytosine methylation is a DNA modification with important regulatory functions in eukaryotes. In flowering plants, sexual reproduction is accompanied by extensive DNA demethylation, which is required for proper gene expression in the endosperm, a nutritive extraembryonic seed tissue. Endosperm arises from a fusion of a sperm cell carried in the pollen and a female central cell. Endosperm DNA demethylation is observed specifically on the chromosomes inherited from the central cell in Arabidopsis thaliana, rice, and maize, and requires the DEMETER DNA demethylase in Arabidopsis. DEMETER is expressed in the central cell before fertilization, suggesting that endosperm demethylation patterns are inherited from the central cell. Down-regulation of the MET1 DNA methyltransferase has also been proposed to contribute to central cell demethylation. However, with the exception of three maize genes, central cell DNA methylation has not been directly measured, leaving the origin and mechanism of endosperm demethylation uncertain. Here, we report genome-wide analysis of DNA methylation in the central cells of Arabidopsis and rice-species that diverged 150 million years agoas well as in rice egg cells. We find that DNA demethylation in both species is initiated in central cells, which requires DEMETER in Arabidopsis. However, we do not observe a global reduction of CG methylation that would be indicative of lowered MET1 activity; on the contrary, CG methylation efficiency is elevated in female gametes compared with nonsexual tissues. Our results demonstrate that locus-specific, active DNA demethylation in the central cell is the origin of maternal chromosome hypomethylation in the endosperm.</P>

      • KCI등재

        Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

        Park, Kyunghyuk,Frost, Jennifer M.,Adair, Adam James,Kim, Dong Min,Yun, Hyein,Brooks, Janie S.,Fischer, Robert L.,Choi, Yeonhee Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.10

        The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

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