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Isolation and Characterization of Eukaryotic Translation Initiation Factor 5A (eIF-5A) from Potato (감자로부터 Eukaryotic Translation Initiation Factor 5A (elF-5A) 유전자의 동정 및 발현 분석)

  • 인준교;신동호;최관삼;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.283-287
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    • 2001
  • Differential display based on PCR was employed to identify genes expressed during tuber-developing stage of potato (Solanum tuberosum L. cv. Irish Cobbler). An eukaryotic initiation factor 5A (eIF-5A) clone isolated from a cDNA library constructed with developing micro-tuber using a probe of PCR fragment. We isolated three positive clones and ore of them contained open reading frame. This clone revealed high sequence similarity to tomato eIF 5A cDNA. At the DNA level, there is 94.8% identity with the tomato eIF-5A4, whereas at the protein level there is a high identity with 97.5%. The potato eIF 5A clone is 716 bp in length and contains a single open reading frame from 57 to 539 bp, a 56 bp 5'-untranslated region and a 177 bp 3'-untranslated region. The deduced protein composed of 160 amino acid residues, with a predicted molecular mass of 17.4 kD and an estimated pl of 5.5. The sequence of 12 (STSKTGKHGHAK) amino acids among eIF-5A proteins is perfectly conserved from yeast to human. That sequence in potato eIF-5A protein is also conserved at position 46 to 57 amino acid. This region embeds the post-translational modification site of the lysine residue (at the seventh K) to hypusine that is crucial to eIF-5A activity. The northern blot analysis of eIF5A has shown abundant expression, mainly in flower organs (stamen, ovary, petal, sepal), fruit and stolen.

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A Fact-finding Study on the Gravesites on the Viewpoint of Landuse in Goryeong Area, Gyeongbug (토지이용면(土地利用面)에서 본 경북(慶北) 고령지역(高靈地域)의 묘지실태(墓地實態)에 관(關)한 조사연구(調査硏究))

  • Kim, Jung-Kon;Jung, Yeun-Tae;Son, Il-Soo;Yoon, El-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.4
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    • pp.315-320
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    • 1987
  • The survey was carried out in the Goryeong area in Gyeongbug province to point out the hinderances of grave increment on the development of hilly to mountainous area and to give basic data to improve the entombment custom. The total extent of gravesite in the area was mapped 148ha which were 0.4% of total area and 1.8% of arableland respectively. Scattering of graves might be hindered utilization of the mountainous area. Out of the total gravesite, about 60% was situated on the areas of 30-60% slopes, and the occupation extent in average was about $89m^2$ per grave which was wider than that of the national average $52m^2$. The soils occupied by gravas ware characterizod by having recommendable properties about 73% for grassland, about 17% for woodland and 8% for orchards.

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Modulation of Escherichia coli RNase E. Action by RraAS2, a Streptomyces coelicolor Ortholog of RraA (Streptomyces coelicolor의 RraA 동족체인 RraAS2에 의한 Escherichia coli RNase E 활성조절)

  • Ahn, Sang-Mi;Shin, Eun-Kyoung;Yeom, Ji-Hyun;Lee, Kang-Seok
    • Korean Journal of Microbiology
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    • v.44 no.2
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    • pp.93-97
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    • 2008
  • RraA is a recently discovered protein inhibitor that regulates the enzymatic activity of RNase E, which plays a major role in the decay and processing of RNAs in Escherichia coli. It has also been shown to regulate the activity of RNase ES, a functional Streptomyces coelicolor ortholog of RNase E, which has 36% identity to the amino-terminal region of RNase E. There are two open reading frames in S. coelicolor genome that can potentially encode proteins having more than 35.4% similarity to the amino acid sequence of RraA. DNA fragment encoding one of these RraA orthologs, designated as RraAS2 here, was amplified and cloned in to E. coli vector to test whether it has ability to regulate RNase E activity in E. coli cells. Co-expression of RraAS2 partially rescued E. coli cells over-producing RNase E from growth arrest, although not as efficiently as RraA, induced by the increased ribonucleolytic activity in the cells. The copy number of ColEl-type plasmid in these cells was also decreased by 14% compared to that in cells over-producing RNase E only, indicating the ability of RraAS2 to inhibit RNase E action on RNA I. We observed that the expression level of RraAS2 was lower than that of RraA by 4.2 folds under the same culture condition, suggesting that because of inefficient expression of RraAS2 in E. coli cells, co-expression of RraAS2 was not efficiently able to inhibit RNase E activity to the level for proper processing and decay of all RNA species that is required to restore normal cellular growth to the cells over-producing RNase E.

A Study on Estimation of Design Tidal level Considering Sea Level Change in the Korean Peninsula (한반도의 해수면 상승을 고려한 설계조위 산정에 관한 연구)

  • Choo, Tai Ho;Sim, Su Yong;Yang, Da Un;Park, Sang Jin;Kwak, Kil Sin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.4
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    • pp.464-473
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    • 2016
  • The air temperatures of the coast and inland are rising due to an increase in carbon dioxide emissions and abnormal climate phenomena caused by global warming, El Nino, La Nina and so on. The sea levels of the Earth are rising by approximately 2.0 mm per year (global average value) due to the thermal expansion of sea water, melting of glaciers and other causes by global warming. On the other hand, when it comes to designing a hydraulic structure or coastal hydraulic structure, the standard of the design water level is decided by analyzing four largeness tide values and a harmonic constant with the observed tidal water level or simulating numerical model. Therefore, the design tidal water level needs to consider an increasing speed of the seawater level, which corresponds to the design frequency. In the present study, the observed tidal water levels targeting 46 tidal stations operated by the Korea Hydrographic and Oceanographic Administration (KHOA) from the beginning of observations to 2015 per hour were collected. The variation of the monthly and yearly and increasing ratio were performed and divided into 7 seas, such as east and west part of the Southern Sea, south part and middle of the East Sea, south part and middle of the Western Sea, and Jeju Sea. The current study could be used to determine the cause of local seawater rises and reflect the design tidal water level as basic data.

Size Distribution and Temperature Dependence of Magnetic Anisotropy Constant in Ferrite Nanoparticles

  • Yoon, Sunghyun
    • Proceedings of the Korean Magnestics Society Conference
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    • 2012.11a
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    • pp.104-105
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    • 2012
  • The temperature dependence of the effective magnetic anisotropy constant K(T) of ferrite nanoparticles is obtained based on the measurements of SQUID magnetometry. For this end, a very simple but intuitive and direct method for determining the temperature dependence of anisotropy constant K(T) in nanoparticles is introduced in this study. The anisotropy constant at a given temperature is determined by associating the particle size distribution f(r) with the anisotropy energy barrier distribution $f_A(T)$. In order to estimate the particle size distribution f(r), the first quadrant part of the hysteresis loop is fitted to the classical Langevin function weight-averaged with the log?normal distribution, slightly modified from the original Chantrell's distribution function. In order to get an anisotropy energy barrier distribution $f_A(T)$, the temperature dependence of magnetization decay $M_{TD}$ of the sample is measured. For this measurement, the sample is cooled from room temperature to 5 K in a magnetic field of 100 G. Then the applied field is turned off and the remanent magnetization is measured on stepwise increasing the temperature. And the energy barrier distribution $f_A(T)$ is obtained by differentiating the magnetization decay curve at any temperature. It decreases with increasing temperature and finally vanishes when all the particles in the sample are unblocked. As a next step, a relation between r and $T_B$ is determined from the particle size distribution f(r) and the anisotropy energy barrier distribution $f_A(T)$. Under the simple assumption that the superparamagnetic fraction of cumulative area in particle size distribution at a temperature is equal to the fraction of anisotropy energy barrier overcome at that temperature in the anisotropy energy barrier distribution, we can get a relation between r and $T_B$, from which the temperature dependence of the magnetic anisotropy constant was determined, as is represented in the inset of Fig. 1. Substituting the values of r and $T_B$ into the $N{\acute{e}}el$-Arrhenius equation with the attempt time fixed to $10^{-9}s$ and measuring time being 100 s which is suitable for conventional magnetic measurement, the anisotropy constant K(T) is estimated as a function of temperature (Fig. 1). As an example, the resultant effective magnetic anisotropy constant K(T) of manganese ferrite decreases with increasing temperature from $8.5{\times}10^4J/m^3$ at 5 K to $0.35{\times}10^4J/m^3$ at 125 K. The reported value for K in the literatures is $0.25{\times}10^4J/m^3$. The anisotropy constant at low temperature region is far more than one order of magnitude larger than that at 125 K, indicative of the effects of inter?particle interaction, which is more pronounced for smaller particles.

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Characterization of Homocysteine ${\gamma}$-Lyase from Submerged and Solid Cultures of Aspergillus fumigatus ASH (JX006238)

  • El-Sayed, Ashraf S.;Khalaf, Salwa A.;Aziz, Hani A.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.499-510
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    • 2013
  • Among 25 isolates, Aspergillus fumigatus ASH (JX006238) was identified as a potent producer of homocysteine ${\gamma}$-lyase. The nutritional requirements to maximize the enzyme yield were optimized under submerged (SF) and solid-state fermentation (SSF) conditions, resulting in a 5.2- and 2.3-fold increase, respectively, after the last purification step. The enzyme exhibited a single homogenous band of 50 kDa on SDS-PAGE, along with an optimum pH of 7.8 and pH stability range of 6.5 to 7.8. It also showed a pI of 5.0, as detected by pH precipitation with no glycosyl residues. The highest enzyme activity was obtained at $37-40^{\circ}C$, with a $T_m$ value of $70.1^{\circ}C$. The enzyme showed clear catalytic and thermal stability below $40^{\circ}C$, with $T_{1/2}$ values of 18.1, 9.9, 5.9, 3.3, and 1.9 h at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. Additionally, the enzyme $K_r$ values were 0.002, 0.054, 0.097, 0.184, and 0.341 $S^{-1}$ at $30^{\circ}C$, $35^{\circ}C$, $40^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$, respectively. The enzyme displayed a strong affinity to homocysteine, followed by methionine and cysteine when compared with non-S amino acids, confirming its potency against homocysteinuria-related diseases, and as an anti-cardiovascular agent and a specific biosensor for homocysteinuria. The enzyme showed its maximum affinity for homocysteine ($K_m$ 2.46 mM, $K_{cat}\;1.39{\times}10^{-3}\;s^{-1}$), methionine ($K_m$ 4.1 mM, $K_{cat}\;0.97{\times}10^{-3}\;s^{-1}$), and cysteine ($K_m$ 4.9 m M, $K_{cat}\;0.77{\times}10^{-3}\;s^{-1}$). The enzyme was also strongly inhibited by hydroxylamine and DDT, confirming its pyridoxal 5'-phosphate (PLP) identity, yet not inhibited by EDTA. In vivo, using Swiss Albino mice, the enzyme showed no detectable negative effects on platelet aggregation, the RBC number, aspartate aminotransferase, alanine aminotransferase, or creatinine titer when compared with negative controls.

A clinical study of the effect of Nd:YAG laser irradiation on inflamed gingiva (염증성 치은에 대한 Nd:YAG laser 조사효과에 관한 임상적 연구)

  • Cho, Hyung-Soo;Kim, Hyun-Sup;Lim, Kee-Jung;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.531-541
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    • 1996
  • Periodontal disease is characterized by destruction of supporting tissues caused by invasion of plaque bacteria and defense mechanism of host. Many dentists are very interested in laser therapy on various intraoral soft tissue lesions including inflammatory periodontal pocket. In order to determine the therapeutic effect of intrapocket irradiation of a pulsed- Nd : YAG laser on the inflammatory periodontal pockets, bilateral 60 teeth with 4-6mm in probing pocket depth and gingival inflammation were selected and evaluated by sulcus bleeding index(SBI), and plaque index(pI) for baseline record. Intrapocket irradiation($300{\mu}m$ fiber optic, I.5W power, for 2 min.) of a pulsed-Nd : YAG laser(EL.EN.EN060, Italy) was applied on half of them. As the control group, the same procedure except power-off was repeated on the contralateral 30 teeth. At 1-, 2-, 3-, and 4-week after intrapocket manipulation, every tooth was reevaluated by the same clinical indices. And the difference between the lased group and control group was statistically analyzed by paired t-test and Chi-square test in Microstat program. Following results were obtained: 1. Until I-week and 2-week after intrapocket manipulation, SBI was lowered in both lased group and control group, compared to baseline SBI, but from 3-week after, the recovering tendency toward baseline was noted, and at only 2-week after, the number of teeth showing lowered SBI was significantly more in lased group than in control group(p<0.05). 2. PI of both lased group and control group was lowered through whole experimental period from I-week to 4-week after, compared to baseline PI(p<0.05), but there was no significant difference between lased group and control group(p>0.1). The results suggest that intrapocket irradiation of a pulsed-Nd:YAG laser may lead somewhat remission of gingival inflammation, but for more favorable therapeutic result the thorough root planing should be necessarily accompanied with gingival curettage.

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CHEMICAL DEGRADATION OF LIGHT CURED COMPOSITE RESINS IN NAOH SOLUTION (NaOH 용액내에서의 광중합형 복합레진의 화학적 분해)

  • Kim, Jung-Ran;Jeong, Byung-Cho;Yang, Kyu-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.3
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    • pp.469-477
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    • 2002
  • One of the major deficiencies of composite restorative resins is their insufficient resistance to wear. Of the multitude of factors that have been associated with wear, subsurface degradation within the restoration is considered to be one. The aim of this study was to evaluate the resistance to degradation of four commercial composite resins in an alkaline solution. The brands studied were Z100(3M), Clearfil AP-X(Kuraray), Tetric Ceram(Vivadent), Aelit flo(Bisco). Resistance to degradation was evaluated on the basis of the following parameters: (a) mass loss(%) - determined from pre-and post-exposure specimen weights: (b) Si loss(ppm) - obtained from ICP-AE analysis of solution exposed to specimens; and (c) degradation depth(${\mu}m$) - measured microscopically (SEM) from polished circular sections of exposed specimens. The results were as follows: 1. The sequence of the mass loss was in ascending order by AE, EL, TC, Z100. There was statistically significant difference of mass loss between AE, CL group and TC, Z100 group(p<0.05). 2. The sequence of the degree of degradation layer depth was in ascending order by AE, CL, TC, Z100. But there was no statistically significant difference of degree of degradation layer depth between AE and CL(p<0.05). 3. For the Si concentration, Z100 was the highest of all. 4. The correlation coefficient between mass loss and degradation depth was relatively high(r=0.71 p<0.05).

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TYPE II DENTINOGENESIS IMPERFECTA : CASE REPORT (Type II 상아질형성부전증의 임상 증례)

  • Kim, Chi-Hyun;Lee, Jae-Ho;Choi, Byung-Jai;Lee, Chong-Gap
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.4
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    • pp.654-660
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    • 2001
  • Dentinogenesis imperfecta is an example of an inheritable dentinal defect originating during the histodifferentiation stage of tooth development, with involvement of the primary and permanent teeth. Shields, Bixler and El-Kafrawy proposed three types of Dentinogenesis imperfecta : Type I, II, III. Witkop reported a prevalence of 1 in 8000 with the trait, and no significant difference between male and female. Affected teeth have red-brown discoloration often with distinctive wearness of occlusal surface of posterior teeth and incisal surface of anterior teeth. Once enamel seperated from underlying defective dentin, the dentin demonstrates significantly acclerated attrision. Radiographically, the teeth have thin roots, bulbous crown, cervical constriction, and obliteration of the root canals and pulp chambers. In primary dentition periapical lesions or multiple root fractures are often observed. In successive generations the phenotypes of discoloration and wearness of teeth occurred, and one of the patient's subships, 10 year-old sister, showed general discoloration of her teeth and mild wearness. In this case, a 4 year-old male reported to the Yonsei University Pedodontics clinic, with a chief complaint of discolored teeth. The teeth showed generally yellowish-brown discoloration and moderate wearness. In radiographic features, obliteration of pulp, bulbous crown, and short roots were observed. It was diagnosed as Dentinogenesis imperfecta. The posterior teeth were restored with Stainless Steel Crown, and defective incisors including left upper primary central incisor which was extracted due to a root fracture with Open-faced Stainless Steel crown.

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Bacteriological Characteristics of Unidentified Vibrio sp., Hemolysin Producer Isolated from Brackish Water -2. Bacteriological Characteristics of Vibrio sp. E10 Similar to Vibrio mimicus- (기수에서 분리된 용혈독소를 생산하는 미분류 Vibrio sp.의 세균학적 특징 -2. Vibrio mimicus와 유사한 Vibrio sp. E10의 세균학적 특성-)

  • Kim Young Man;Yu Hong Sik;Oh Hee Kyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.545-550
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    • 2002
  • A hemolysin producing bacterial strain which belong to Vibrio species was isolated from the Kum River estuary. In the process of identification, the strain did not show characteristics of known Vibrio species; thus, the strain was designated as Vibrio sp, E10 (V. kunsan) tentatively and further identification study was carried out by comparing its bacteriological characteristics. Morphologically Vibrio sp, E10 was comma shaped rod with a polar flagellium. Clear hemolysis zones were observed with the strain against human and sheep blood agar. Hemollytic toxicity was confirmed by strong vascular Permeability and fatal toxicity against mouse was also observed. Therefore the strain was a pathogenic vibrio. Growth conditions for Vibrio sp. E10 were ranged salinity of 0$\~$$4.5\%$, pH of 6.2$\~$9.2, temperature of 14$\~$42$^{\circ}C$, respectively, 16S rDNA partial sequence of Vibrio sp, E10 showed $99\%$ homology with dozens of V. cholerae species including V, cholerae El Tor N16961 and V, snmisnfus ATCC 33653T. This strain belonged to Proteobacteria; gamma subdivision; Vibrionacea: Vibrio. But, among knorn Vibrio species no identical styains were found when using automatic bacteria identification system ($MicroLog^{TM}$system, release 4.0, Biolog Inc., USA) which evaluated the ability of metabolizing 95 kinds of carbon and nitrogen sources. Vibrio sp, E10 showed 18 and 11 different responses as compared to V. mimicus and V, cholerae, respectively.