Kim, Sung-Kuk;Wee, Sung-Mo;Chang, Jong-Soo;Kwon, Taeg-Kyu;Min, Do-Sik;Lee, Young-Han;Suh, Pann-Ghill
BMB Reports
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v.37
no.6
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pp.720-725
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2004
A number of signaling molecules contain small pleckstrin homology (PH) domains capable of binding phosphoinositides or proteins. Phospholipase C (PLC)-${\gamma}1$ has two putative PH domains, an $NH_2$-terminal (PH1) and a split PH domain ($nPH_2$ and $cPH_2$). We previously reported that the split PH domain of PLC-${\gamma}1$ binds to phosphatidylinositol 4-phosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate (PI(4,5)$P_2$) (Chang et al., 2002). To identify the amino acid residues responsible for binding with PI(4)P and PI(4,5)$P_2$, we used site-directed mutagenesis to replace each amino acid in the variable loop-1 (VL-1) region of the PLC-${\gamma}1$$nPH_2$ domain with alanine (a neutral amino acid). The phosphoinositide-binding affinity of these mutant molecules was analyzed by Dot-blot assay followed by ECL detection. We found that two PLC-${\gamma}1$ nPH2 domain mutants, P500A and H503A, showed reduced affinities for phosphoinositide binding. Furthermore, these mutant PLC-${\gamma}1$ molecules showed reduced PI(4,5)$P_2$ hydrolysis. Using green fluorescent protein (GFP) fusion protein system, we showed that both $PH_1$ and $nPH_2$ domains are responsible for membrane-targeted translocation of PLC-${\gamma}1$ upon serum stimulation. Together, our data reveal that the amino acid residues $Pro^{500}$ and $His^{503}$ are critical for binding of PLC-${\gamma}1$ to one of its substrates, PI(4,5)$P_2$ in the membrane.
Zinc finger (ZNF) proteins play a critical role in cell growth, proliferation, apoptosis, and intracellular signal transduction. In this paper, we cloned and characterized a novel human KRAB-related zinc finger gene, ZNF425, which encodes a protein of 752 amino acids. ZNF425 is strongly expressed in the three month old human embryos and then is almost undetectable in six month old embryos and in adult tissues. An EGFP-ZNF425 fusion protein can be found in both the nucleus and the cytoplasm. ZNF425 appears to act as a transcription repressor. Over-expression of ZNF425 inhibits the transcriptional activities of SRE, AP-1, and SRF. Deletion analysis indicates that the C2H2 domain is the main region responsible for the repression. Our results suggest that the ZNF425 gene is a new transcriptional inhibitor that functions in the MAPK signaling pathway.
Rewards or penalties become informative only when contingent on an immediately preceding response. Our goal was to determine if the brain responds differently to motivational events depending on whether they provide feedback with the contingencies effective for learning. Event-related fMRI data were obtained from 22 volunteers performing a visuomotor categorical task. In learning-condition trials, participants learned by trial and error to make left or right responses to letter cues (16 consonants). Monetary rewards (+500) or penalties (-500) were given as feedback (learning feedback). In random-condition trials, cues (4 vowels) appeared right or left of the display center, and participants were instructed to respond with the appropriate hand. However, rewards or penalties (random feedback) were given randomly (50/50%) regardless of the correctness of response. Feedback-associated BOLD responses were analyzed with ANOVA [trial type (learning vs. random) x feedback type (reward vs. penalty)] using SPM8 (voxel-wise FWE p < .001). The right caudate nucleus and right cerebellum showed activation, whereas the left parahippocampus and other regions as the default mode network showed deactivation, both greater for learning trials than random trials. Activations associated with reward feedback did not differ between the two trial types for any brain region. For penalty, both learning-penalty and random-penalty enhanced activity in the left insular cortex, but not the right. The left insula, however, as well as the left dorsolateral prefrontal cortex and dorsomedial prefrontal cortex/dorsal anterior cingulate cortex, showed much greater responses for learning-penalty than for random-penalty. These findings suggest that learning-penalty plays a critical role in learning, unlike rewards or random-penalty, probably not only due to its evoking of aversive emotional responses, but also because of error-detection processing, either of which might lead to changes in planning or strategy.
In this paper, to elucidate the further mechanisms of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced growth arrest, we investigated the effect of MNNG on cell cycle and proliferation in U937 cells, a p53-null human myeloid leukemia cell line. It was found that MNNG causes an arrest at the G2/M phase of the cell cycle and induces apoptosis, which is closely correlated to inhibition of cyclin B1 and cyelin-dependent kinase (Cdk) 2-associated kinase activities. MNNG treatment in. creased protein and mRNA levels of the Cdk inhibitor p21(WAF1/CIP1), and activated the reporter construct of a p21 promoter. By using p21 promoter deletion constructs, the MNNG-responsive element was mapped to a region between 113 and 61 relative to the transcription start site. These data indicate that in U937 cells MNNG can circumvent the loss of wild-type p53 function and induce critical downstream regulatory events leading to transcriptional activation of p21. Present results indicate that the p53-independent up-regulation of p21 by MNNG is likely responsible for the inhibition of cyclin/Cdk complex kinase activity rather than the down-regulation of cyclins and Cdks expression. These novel phenomena have not been previously described and provide important new insights into the possible biological effects of MNNG.
In this study MeV Si self ion implantations were done to reveal the intrinsic behavior of defect formation by excluding the possibility of chemical interactions between substrate atoms and dopant ones. Self implantations were conducted using Tandem Accelerator with energy ranges from 1 to 3 MeV. Defect formation by high energy ion implantation has a significant characteristics in that the lattice damage is concentrated near Rp and isolated from the surface. In order to investigate the energy dependence on defect formation, implantation energies were varied from 1 to 3 MeV under a constant dose of $1{\times}10^{15}/cm^2$. RBS channe!ed spectra showed that the depth at which as-implanted damaged layer formed increases as energy increases and that near surface region maintains better crystallinity as energy increases. Cross sectional TEM results agree well with RBS ones. In a TEM image as-implanted damaged layer appears as a dark band, where secondary defects are formed upon annealing. In the case of 2 MeV $Si^+$ self implantation a critical dose for the secondary defect formation was found to be between $3{\times}10^{14}/cm^24$ and $5{\times}10^{14}/cm^2$. Upon annealing the upper layer of the dark band was removed while the bottom part of the dark band did not move. The observed defect behavior by TEM was interpreted by Monte Carlo computer simulations using TRIM-code. SIMS analyses indicated that the secondary defect formed after annealing gettered oxygen impurities existed in silicon.
The geometrical shape of a plow bottom may be the most important factor affecting the performance of a plow for a given soil and operating conditions. There are various designs of the Jaenggi (Korean plow) available commercially, which may be different from each other and from the plow (Western plow) in respect to the shape and performance. This study was intended to investigate the geometrical characteristics of Jaenggi. The coordinate digitizer equipped with 3 potentiometers was designed and manufactured for measurement of the shape of curved plane of moldboard and share. The digitizer was connected to a microcomputer having the data acquisition system. This device was used to analyze the plow bottoms of 5 differently-made Jaenggis and one cylindrical plow. The results of the study are summarized as follows: 1. It was possible to measure easily and quickly the curved plane of plow bottom and to plot the view on three major plans using the coordinate digitizer electrically connected to a microcomputer system. 2. The shape of five Jaenggi bottoms analyzed could be characterized by the cutting angle having the range of $33-42^{\circ}$, the maximum share-lift angle of $41-50^{\circ}$, and the setting angle of moldboard wing of $46-70^{\circ}$. The most critical difference of the shape factors between the Jaenggi and the plow was found in the maximum share-lift angle, the former was more than twice as much as the latter. 3. The analysis of the shape of Jaenggi bottoms showed that the share projections on 3 major plans had a varied triangle, which was quite different from that of plow bottom. Especially, it was analyzed that the shape of furrow slice for the Jaenggi had a skewed rectangle, leaving a considerable height of the ridge at the furrow bottom. 4. The dihedral angle was similar range of $30-85^{\circ}$ for the all bodies investigated, but the directional angle was somewhat different from each other. The difference in directional angle was $5^{\circ}$ for the plow, $20^{\circ}$ for the Jaenggi A and $30^{\circ}$ for the Jaenggi B. 5. Area of the spherical representation region was 0.0328 for the plow, 0.1194 for the Jaenggi A and 0.1716 for the Jaenggi B. This may indicate that the plow came close to a working surface and the Jaenggi A and the Jaenggi B departed from a working surface to a somewhat greater extent.
Kim, Jae-Yoon;Kim, Dae-Yeon;Jung, Je-Hyeong;Hong, Min-Jeong;Heo, Hwa-Young;Johnson, Jerry W.;Kim, Tae-Ho;Seo, Yong-Weon
Journal of Crop Science and Biotechnology
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v.10
no.1
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pp.50-56
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2007
Barley S-adenosylmethionine synthetase1 gene, which was differentially expressed in seed development of extra early barley, was regulated by the phytohormones and abiotic stresses. In order to identify the regulation regions which were involved in transcriptional control of the phytohormones and abiotic stresses, we isolated 1459 bp fragment of HvSAMS1 gene promoter using genome walking strategy and deletion series were constructed. Deleted upstream fragments(-1459, -1223, -999, -766, -545, -301 bp) were fused to the GUS reporter gene and evaluated via Agrobacterium-mediated transient expression assay. Increased GUS activity of HvSMAS1 promoter -301/GUS construct under each of NaCl, $GA_3$, ABA and ethylene application was found. However, GUS activity was negligible in the leaves transformed with the HvSMAS1 promoter(-1459, -1223, -999, -766 and -545)/GUS constructs. No significant induction of GUS activity was observed for the ethionine and spermidine treatments. In order to locate promoter sequence of the HvSAMS1 gene that was critical for the activation of gene expression, deletion and addition promoter derivatives(+, includes 43 bp of 5' ORF) of the HvSAMS1 gene fused to the GUS reporter gene were applied. The tobacco leaves which harbored the additional HvSAMS1 promoter(-1459+, -1459 to -546, -545+ and -301+)/GUS construct did not significantly induce GUS activity as compared to the HvSAMS1 promoter(-1459, -545 and -301)/GUS constructs under each of NaCl, ABA and $GA_3$ treatment. However, the GUS activity was high in the tobacco leaves which harboring the -211 to -141 regions of the HvSAMS1 promoter. This result suggested that HvSAMS1 gene expression might be regulated by this region(from -211 to -141).
Journal of Korean Society for Geospatial Information Science
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v.19
no.3
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pp.57-66
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2011
The purpose of this study is to research into a method of selecting the subject parcel with a change in the category of land given surveying the land alteration state focusing on the present state of road in the government-owned and public land by using the fuzzy membership function and GIS spatial analysis. It selected the old town center of Incheon Jung-gu, and the new downtown & the forest land of Gyeyang-gu as the research subject region, and carried out GIS spatial analysis on a serial cadastral map, urban planning road layer of Korea Land Information System, practical width of road layer of Road Name Address Management System & cadastral data base, and then calculated the suitable index for the subject parcel with a change in the category of land by using the fuzzy membership function with having the critical value as the area ratio of each parcel on a serial cadastral map that was incorporated into road layer or practical width of road layer. It finally selected the parcel, which is different in land category from the real land usage, as the final subject parcel for altering land category, by using the screen of visualizing the suitable index and the aerial ortho photograph. As a result of the final selection, the fuzzy GIS spatial analysis method, which was suggested in this study, is judged to be efficient in the selection period and the methodology compared to the existing manual method. It could be confirmed to be more suitable method for downtown than forest land and for the new downtown than the old town center.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.33
no.2
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pp.114-120
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2007
The augmentation of soft tissue defects is one of the critical problems in the oral and maxillofacial surgery. Various types of graft materials, both autologous and non-autologous, have been used for the augmentation of soft tissue in the facial region. However, it is not easy to choose an ideal material for soft tissue augmentation because each has its advantages and disadvantages. An ideal graft material should meet the following criteria : it should not leave a scar at the area from which it was taken; should have less likelihood of causing infection; should feel natural after implanted; and should be not absorbed. Among the materials meeting these criteria, human dermis and artificial dermis are commonly used for clinical purposes. The present study was aimed to investigate and compare the resorption rate and the histological change following the use of the autologous dermis, the human homogenous dermis $Alloderm^{(R)}$, and the artificial dermis $Terudermis^{(R)}$ to reconstruct the soft tissue defect. Twenty mature rabbits of either sex, weighing about 2 ㎏, were used. Each rabbit was transplanted with the autologous dermis, $Alloderm^{(R)}$, and $Terudermis^{(R)}$ size $1{\times}1-cm$ at the space between the external abdominal oblique muscle and the external abdominal oblique fascia. They were then divided into 4 groups (n=5 each) according to the time elapsed after the surgery: 1, 2, 4, and 8 weeks. The resorption rate was calculated by measuring the volume change before and after the transplantation, and H-E stain was preformed to observe the histological changes. The resorption rate after 8 weeks was 21.5% for the autologous dermis, 16.0% $Alloderm^{(R)}$, and 36.4% $Terudermis^{(R)}$, suggesting that $Alloderm^{(R)}$ is the most stable while $Terudermis^{(R)}$ is the most unstable. In microscopic examinations, the autologous dermis graft was surrounded by inflammatory cells and showed foreign body reactions. The epidermal inclusion cyst was observed in the autologous dermis graft. $Terudermis^{(R)}$ and $Alloderm^{(R)}$ demonstrated neovascularization and the progressive growth of new fibroblast. The results suggest that $Terudermis^{(R)}$ and $Alloderm^{(R)}$ can be availably for substituting the autologous dermis.
Attentional processes facilitate cognitive and behavioral performance in several ways. Attention serves to reduce the amount of information to receive. Attention enables humans to direct themselves to appropriate aspects of external environmental events and internal operations. Attention facilitates the selection of salient information and the allocation of cognitive processing appropriate to that information. Attention is not a unitary process that can be localized to a single neuroanatomical region. Before the cortical registration of sensory information, activation of important subcortical structures occurs, which is called as an orienting response. Once sensory information reaches the sensory cortex, a large number of perceptual processes occur, which provide various levels of perceptual resolution of the critical features of the stimuli. After this preattentional processing, information is integrated within higher cortical(heteromodal) systems in inferior parietal and temporal lobes. At this stage, the processing characteristics can be modified, and the biases of the system have a direct impact on attentional selection. Information flow has been traced through sensory analysis to a processing stage that enables the new information to be focused and modified in relation to preexisting biases. The limbic and paralimbic system play significant roles in modulating attentional response. It is labeled with affective salience and is integrated according to ongoing pressures from the motivational drive system of the hypothalamus. The salience of information greatly influences the allocation of attention. The frontal lobe operate response selection system with a reciprocal interaction with both the attention system of the parietal lobe and the limbic system. In this attentional process, the search with the spatial field is organized and a sequence of attentional responses is generated. Affective, motivational and appectitive impulses from limbic system and hypothalamus trigger response intention, preparation, planning, initiation and control of frontal lobe on this process. The reticular system, which produces ascending activation, catalyzes the overall system and increases attentional capacity. Also additional energetic pressures are created by the hypothalamus. As psychophysiological measurement, skin conductance, pupil diameter, muscle tension, heart rate, alpha wave of EEG can be used. Event related potentials also provide physiological evidence of attention during information process. NI component appears to be an electrophysiological index of selective attention. P3 response is developed during the attention related to stimulus discrimination, evaluation and response.
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