• Title/Summary/Keyword: Biological sequence

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Application of Molecular Biological Technique for Development of Stability Indicator in Uncontrolled Landfill (불량매립지 안정화 지표 개발을 위한 분자생물학적 기술의 적용)

  • Park, Hyun-A;Han, Ji-Sun;Kim, Chang-Gyun;Lee, Jin-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.2
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    • pp.128-136
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    • 2006
  • This study was conducted for developing the stability parameter in uncontrolled landfill by using a biomolecular investigation on the microbial community growing through leachate plume. Landfill J(which is in Cheonan) and landfill T(which is in Wonju) were chosen for this study among a total of 244 closed uncontrolled landfills. It addressed the genetic diversity of the microbial community in the leachate by 165 rDNA gene cloning using PCR and compared quantitative analysis of denitrifiers and methanotrophs with the conventional water quality parameters. From the BLAST search, genes of 47.6% in landfill J, and 32.5% in landfill T, respectively, showed more than 97% of the similarity where Proteobacteria phylum was most significantly observed. It showed that the numbers of denitrification genes, i.e. nirS gene and cnorB gene in the J site are 7 and 4 times higher than those in T site, which is well reflecting from a difference of site closure showing 7 and 13 years after being closed, respectively. In addition, the quantitative analysis on methane formation gene showed that J1 spot immediately bordering with the sources has the greatest number of methane formation bacteria, and it was decreased rapidly according to distribute toward the outer boundary of landfill. The comparative investigation between the number of genes, i.e. nirS gene, cnorB gene and MCR gene, md the conventional monitoring parameters, i.e. TOC, $NH_3-N,\;NO_3-N,\;NO_2-N,\;Cl^-$, alkalinity, addressed that more than 99% of the correlation was observed except for the $NO_3-N$. It was concluded that biomolecular investigation was well consistent with the conventional monitoring parameters to interpret their influences and stability made by leachate plume formed in downgradient around the uncontrolled sites.

ANALYSIS OF DIFFERENTIAL GENE EXPRESSION IN NORMAL, CYST AND AMELOBLASTOMA CELLS (정상, 낭종 및 법랑아세포종 세포에서의 유전자 발현 차이 분석)

  • Yang, Cheol-Hee;Baik, Byeong-Ju;Yang, Yeon-Mi;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.32 no.1
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    • pp.75-88
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    • 2005
  • Ameloblastoma is the most commonly occurring odontogenic tumor in oral cavity. Although most are benign epithelial neoplasm, they are generally considered to be locally aggressive and destructive, exhibiting a high rate of recurrence. The biological behavior of this neoplasm is a slowly growing, locally invasive tumor without metastasis, therefore malignant neoplasm, changed its histological appearance to carcinoma or showed distant metastasis, is only defined clinically. In this study, we identified the differentially expressed genes(DEGs) in stages under benign or malignant ameloblastoma compared with normal patient using ordered differential display(ODD) reverse transcription(RT)-PCR and $GeneFishing^{TM}$ technology. ODD RT-PCR is rather effective when the investigation of samples containing very small amounts of total RNA must be accomplished. ODD RT-PCR used the means of amplification with anchored T-primer and adaptor specific primer. bearing definite two bases at their 3' ends and so this method could display differential 3'-expressed sequence taqs(ESTs) patterns without using full-length cDNAs. Compared with standard differential display, ODD RT-PCR is more simple and have enough sensitivity to search for molecular markers by comparing gene expression profiles, However, this method required much effort and skill to perform. $GeneFishing^{TM}$ modified from DD-PCR is an improved method for detecting differentially expressed genes in two or more related samples. This two step RT-PCR method uses a constant reverse primer(anchor ACP-T) to prime the RT reaction and arbitrary primer pairs(annealing control primers, ACPs) during PCR. Because of high annealing specificity of ACPs than ODD RT-PCR, the application of $GeneFishing^{TM}$ to DEG discovery generates reproducible, authentic, and long(100bp to 2kb) PCR products that are detectable on agarose gels. Consequently, various DEGs observed differential expression levels on agarose gels were isolated from normal, benign, and malignant tissues using these methods. The expression patterns of the some isolated DEGs through ODD RT-PCR and $GeneFishing^{TM}$ were confirmed by Northern blot analysis and RT-PCR. The results showed that these identified DEGs were implicated in ameloblastoma neoplasm processes. Therefore, the identified DEGs will be further studied in order to be applied in candidate selection for marker as an early diagnosis during ameloblastoma neoplasm processes.

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Evaluation of Antidepressant Drug Effect in a Depressive Animal Model by Proton MR Spectroscopy (양성자 자기공명분광법을 이용한 우울증 동물모델에서의 항우울제 약물 효능 평가)

  • Kim, Sang-Young;Choi, Chi-Bong;Lee, Sung-Ho;Woo, Dong-Cheol;Yoon, Seong-Ik;Hong, Kwan-Soo;Lee, Hyun-Sung;Cheong, Chae-Joon;Jee, Bo-Keun;Hong, Sung-Tak;Kim, Hwi-Yool;Choe, Bo-Young
    • Progress in Medical Physics
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    • v.19 no.2
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    • pp.95-101
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    • 2008
  • In this study, we observed the alteration of choline signal intensity in hippocampus region of the depressive rat model induced by forced swimming test (FST). The purpose of this study was to evaluate the antidepressant efficacy in the depressive animal model using MR spectroscopy. Fourteen experimentally naive male Sprague-Dawley rats weighting $160{\sim}180\;g$ were used as subjects. Drug injection group was exposed to the FST except for control group. The drugs were administered subcutaneously (SC) in a volume equivalent to 2ml/kg. And three injections were administered 23, 5, and 1h before beginning the given test. 1H MR spectra were obtained with use of a point resolved spectroscopy (PRESS) localization sequence performed according to the following parameters: repetition time, 2500 ms; echo time, 144 ms; 512 average; 2048 complex data points; voxel dimensions, $1.5{\times}2.5{\times}2.5\;mm^3$ ; acquisition time, 25min. There were no differences in NAA/Cr and Cho/Cr ratio between the right and the left hippocampus both normal control rats and antidepressant-injected rats. Also, no differences were observed in NAA/Cr and Cho/Cr ratio between the normal control rats and the antidepressant-injected rats both the right and the left hippocampus. In this study, we found the recovery of choline signals in the depressive animal model similar to normal control groups as injecting desipramine-HCl which was antidepressant causing anti-immobility effects. Thus, we demonstrated that MR spectroscopy was able to aid in evaluating the antidepressant effect of desipramine-HCl.

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Identification of the Pig β-1,3-N-acetylglucosaminyltransferase 1 (pB3GNT1) that is Involved in Poly-N-acetyllactosamine (poly-LacNAc) Synthesis (Poly-N-acetyllactosamine (poly-LacNAc) 합성에 관여하는 돼지 β-1,3-N-acetylglucosaminyltransferase I (pB3GNT1) 유전자 동정)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Hochi, Shinichi;Park, Mi-Ryung;Byun, Sung June;Oh, Keon Bong;Yang, Hyeon;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.28 no.4
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    • pp.389-397
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    • 2018
  • The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

Phylogenetic Analysis of Korean Native Aster Plants Based on Internal Transcribed Spacer (ITS) Sequences (ITS 염기서열을 이용한 한국산 참취속 식물의 유연관계분석)

  • Hong, Su-Young;Cho, Kwang-Soo;Yoo, Ki-Oug
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.178-184
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    • 2012
  • This study was carried out to decide ITS (internal transcribed spacer) sequence of some Korean native $Aster$ species and to resolve their relationship among Korean native $Aster$, including $Kalimeris$, $Gymnaster$, $Heteropappus$ genus separated from $Aster$ in a previously study based on the pappus length. We registered 11 ITS sequences of $Aster$ species including $A.$ $glehni$ to GenBank and those sequences were used for the cluster analysis with $Kalimeris$ species. The size of ITS1 was varied from 248 to 256 bp, while ITS2 was varied from 220 to 222 bp. The G + C content of the ITS region ranged from 49.4 to 53.5%. Pairwise comparison results showed that the substitution rate of ITS1 and ITS2 region was 9% and 10%, respectively. $Kalimeris$ sensu strict substitution rate was lower than that of $Aster$ sensu strict species. The strict consensus parsimonious cluster analysis showed $A.$ $tripolium$ is the first branching from the clade and the next is $A.$ $scaber$. The $Kalimeris$ species except for the $A.$ $hispidus$ were grouped into the same clade with high bootstrap value (91%) within $Aster$. $Gymnaster$ and $Heteropappus$ that has been classified by morphological characters were also grouped into broad sense $Aster$ clade. These results implied these three genera could be merged together into $Aster$ based on the ITS sequences.

Gadoteridol's Signal Change according to TR, TE Parameters in T1 Image (T1영상에서 TR, TE 매개변수에 따른 Gadoteridol의 신호강도 변화)

  • Jeong, Hyun Keun;Jeong, Hyun Do;Nam, Ki Chang;Kim, Ho Chul
    • Journal of the Institute of Electronics and Information Engineers
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    • v.52 no.9
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    • pp.117-124
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    • 2015
  • In this paper, we introduce how to control TR, TE physical MR parameters for managing $H_1$ spin's SI(Signal Intensity) which is combined with gadolinium following administration MR agent in T1 effect for diagnostic usefulness. we used MRI phantom made with 0.5 mol Gadoteridol. This phantom was scanned by FSE sequence with different TR, TE parameters. In this study, to make T1 effect, TR was 200, 250, 300, 350, 400, 450, 500, 550, 600 msec. In addition to, TE was 6.2, 12.4, 18.6, 21.6 msec. The results were as follows ; Each RSP(Reaction Starting Point) was 100, 50, 40, 30 mmol in TE 6.2, 12.4, 18.6, 21.6 msec being irrelevant to TR. In MPSI(Max Peak Signal Intensity), 4 mmol was showed in TR 200 msec while peak signal was decreased to low concentration mol in TR 250-600 msec. In terms of RA(Reaction Area), the highest SI was TE 6.2 msec in TR 200-600msec. According to the study, we are able to recognize it is possible to control enhance rates by managing TR and TE of MR parameters; moreover, we expect that enhanced T1 image in MR clinical field can be performed in a practical way with this quantitative data.

From Trauma To growth: Posttraumatic Growth Clock (외상 후 병리에서 성장으로: 외상 후 성장 시계)

  • Lee, Hong-Seock
    • Korean Journal of Cognitive Science
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    • v.27 no.4
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    • pp.501-539
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    • 2016
  • The human mind is a self-evolving system that develops along a multidimensional hierarchical pathway in response to traumatic stimulus. In absence of trauma, a mind integrated in conflict-free state is called monistic. When the monistic mind responses to a traumatic stimulus, a response polarity forms toward stimulus polarity within the mind, turning it into a bipartite structure. Dialectical interaction between the two opposites, originating from their incompatibility, creates a new third polarity in the upper dimension. Thereby, the mind turns into a trinity structure. When the interaction among the three polarities becomes optimized, the plasticity of the mind gets maximized into the "far-from-equilibrium state," and the function of three polarities is synchronized. Through this recalibration, the mind returns back to its monistic structure. If the mind with the recurred monistic structure responds to another traumatic stimulus, this cycle of hierarchical transformation repeats itself in this cyclical and fractal growth process through synchronization of basic trinity system. Applying this concept to the process of post-traumatic growth (PTG), this paper explores how the mind transforms traumatic experiences into PTG and proposes a 'PTG Clock' that shows a fundamental sequence in the development of the human mind. The PTG Clock consists of seven hierarchical phases, and each of the first six phases has two opposite sub-phases: shocked/numbed, feared/intrusive, paranoid/avoidant, obsessional/explosive, dependent/depressive, and meaningless/searching for meaning. The seventh, the synchronization phase, completes one cycle of the mind's transformation, realizing a grand trinity system, where the mind synchronizes its biological, social, and existential dimensions. At that point, the mind becomes more susceptible to not only the stimulus of its own traumatic experience but also the pain of others. Thereby, the PTG Clock sets out on a journey to another cycle of transformation in higher dimensions. The validity of this transformational process for the PTG Clock will be examined by comparing it to Horowitz's theory of stress response syndrome.

Molecular Characterization of Cinnamate 4-Hydroxylase gene in Red Hot Pepper (Capsicum annuum L.) (고추에서 분리한 Cinnamate 4-Hydroxylase 유전자의 분자생물학적 특성)

  • Kim Kye-Won;Ha Sun-Hwa;Cho Kang-Jin;Kim Eun-Ju;Lee Min-Kyung;Yu Jae-Ju;Kim Jong-Guk;Lee Shin-Woo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.167-173
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    • 2005
  • Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.

Production of Poly-3-hydroxybutyrate from Xylose by Bacillus megaterium J-65 (Bacillus megaterium J-65에 의한 xylose로부터 poly-3-hydroxybutyrate 생산)

  • Jun, Hong-Ki;Jin, Young-Hi;Kim, Hae-Nam;Kim, Yun-Tae;Kim, Sam-Woong;Baik, Hyung-Suk
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1625-1630
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    • 2008
  • A microorganism capable of producing high level of poly-3-hydoxybutyrate (PHB) from xylose was isolated from soil. The isolated strain J-65 was identified as Bacillus megaterium based on the morphological, biochemical and molecular biological characteristics. The optimum temperature and pH for the growth of B. megaterium J-65 were $37^{\circ}C$ and 8.0, respectively. The optimum medium composition for the cell growth was 2% xylose, 0.25% $(NH_4)_2SO_4$, 0.3% $Na_2HPO_4{\cdot}12H_2O$, and 0.1% $KH_2PO_4$. The optimum condition for PHB accumulation was same to the optimum condition for cell growth. Copolymer of ${\beta}$-hydroxybutyric and ${\beta}$-hydroxyvaleric acid was produced when propionic acid was added to shake flasks containing 20 g/l of xylose. Fermenter culture was carried out to produce the high concentration of PHB. In batch culture, cell mass was 9.82 g/l and PHB content was 35% of dry cell weight. PHB produced by B. megaterium J-65 was identified as homopolymer of 3-hydoxybutyric acid by GC and NMR.

Effects of Characteristics of Ovarian follicular Fluid and Ant-Inhibin Serum on Steroid Hormone Secretion by Hanwoo Granulosa Cells In Vitro (한우 난소의 Follicular Fluid의 특징과 과립막 세포의 스테로이드호르몬 분비에 대한 Anti-Inhibin Serum의 첨가효과)

  • 성환후;민관식;양병철;노환국;최선호;임기순;장유민;박성재;장원경
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.119-124
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    • 2001
  • This study was performed to investigate the effects of the peptide to carrier ratio on the immune and biological functions to inhibin immunization in Hanwoo. A peptide sequence kom the alpha -subunit (19~32 peptide) of porcine inhibin was synthesized for antigen and conjugated to human serum albumin(HSA) for carrier protein. Anti-inhibin sera(AI) were produced 52 day later from rabbit after injection of inhibin-$\alpha$ -subunit peptide conjugator for antigen with the interval of 2 weeks. Immune-blotting analysis using antibody specific fur inhibin-$\alpha$ subunits revealed that the inhibin was detected at 1.0 cm bovine follicular fluid(bFF). However, each stage of corpus lutea and 0.1 cm of follicular fluid were not detected. The maximal contents of estradiol-17 $\beta$ in Hanwoo ovarian follicular fluid were detected at 2.0 cm of follicular size(diameter), but the mean total contents of these hormone decreased significantly with decreasing diameter of follicles. However, progesterone contents of follicular fluid were high at 1.0 cm of follicle. Progesterone secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) inhibited in 5% bFF and 5% bFF + 5% AI addition group compared with control group. Estradiol-17 $\beta$ secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) increased in 5% AI and 5% AI + 5% bFF addtion group compared with control group. However, the groups added 5% AI were not changed compared to control groups in progesterone and estradiol-17 $\beta$. Taken together, we suggested that inhibin in the mature FF plays a pivotal role on the biosynthesis of steroid hormone of follicular cells during follicular development.

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