• Title/Summary/Keyword: 14-3-$3{\beta}$

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Cloning, Characterization of Pichia etchellsii $\beta-Glucosidase$ II and Effect of Media Composition and Feeding Strategy on its Production in a Bioreactor

  • Sethi Benu;Jain Monika;Chowdhary Manish;Soni Yogesh;Bhatia Yukti;Sahai Vikram;Mishra Saroj
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.1
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    • pp.43-51
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    • 2002
  • The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.

A Study on Disaster Preparedness, Core Competencies and Educational Needs on Disaster Nursing of Nursing Students (간호대학생의 재난준비도, 재난간호 핵심수행능력 및 교육요구도에 대한 연구)

  • Kim, Hee-Jung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.11
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    • pp.7447-7455
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    • 2015
  • This study was aimed to investigate disaster preparedness, core competencies and educational needs on disaster nursing of nursing students and to provide basic data needed for development of disaster nursing educational program. 254 nursing students enrolled in 3 college of nursing in D-city completed questionnaires. The data were collected from November 1, 2014 to November 30, 2014. The average level of core competencies on disaster nursing was 2.76 out of 5 points, which was moderate for the core competencies on disaster nursing. Disaster preparedness was 2.14 out of 5 points, suggesting that they are generally not well prepared for disaster. Factors affecting core competencies on disaster nursing were disaster preparedness, educational needs, grade, and experience of disaster education. It is necessary to develop disaster nursing educational program to reflect the needs of the field in Korea. And Nurse educator needs to develop strategies to prepare their students for disasters. Further research is needed to adequately address this issue.

The Crystal and Molecular Structure of Sodium Sulfisoxazole hexahydrate (Sodium Sulfisoxazole Hexahydrate의 결정 및 분자구조와 수소결합에 관한 연구)

  • Young Ja Park;Chung Hoe Koo
    • Journal of the Korean Chemical Society
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    • v.20 no.1
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    • pp.19-34
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    • 1976
  • The crystal structure of sodium sulfisoxazole hexahydrate, $C_{11}H_{12}N_3O_3SNa{\cdot}6H_2O$,has been determined by X-ray diffraction method. The compound crystallizes in the monoclinic space group $$P2_1}c$$ with a = 15.68(3), b = 7.70(2), c = 17.94(4)${\AA}$, ${\beta}$ = $118(2)^{\circ}$ and Z = 4. A total of 1717 observed reflections were collected by the Weissenberg method with $CuK{\alpha}$ radiation. Structure was solved by heavy atom method and refined by block-diagonal least-squares methods to the R value of 0.14. The conformational angle formed by the S-C(l) bond with that of N(2)-C(7), when the projection in taken along the S-N(2), is $73^{\circ}.$ The benzene ring is planar and makes an angle of $60^{\circ}$ with the plane of the isoxazole ring, which is also planar. The sodium atom has a distorted octahedral coordination of N(l) and five oxygen atoms from hydrate molecules. Sodium sulfisoxazole hexahydrate shows fourteen different hydrogen bondings in the crystal. These are six $O-H{\cdots}O-H bonds, three $O-H{\cdots}O$ bonds, two $O-N{\cdots}N,$ one $N-H{\cdots}O,O-H{\cdots}N,N-H{\cdots}O-H$ bond, with the distances in the range of 2.71 to $3.04{\AA}.$.

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Phytochemical Studies on Paeoniae Radix (2);Phenolic and Related compounds (작약(芍藥)의 성분연구(成分硏究) (2);Phenol성 물질 및 관련화합물들의 분리)

  • Kim, Ju-Sun;Kim, Yoon-Jung;Lee, Joo-Young;Kang, Sam-Sik
    • Korean Journal of Pharmacognosy
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    • v.39 no.1
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    • pp.28-36
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    • 2008
  • From the 70% EtOH extract of Paeonia lactiflora roots (Paeoniaceae), fourteen phenolic and related compounds were isolated. They were identified as ${\alpha}-tocopherol$ (1), dioctylphthalate (2), ${\alpha}-tocospiro$ B (3), paeonol (4), 3,3'-di-O-methylellagic acid(5), 3,4'-di-O-methylellagic acid (6), benzoic acid (7), aromadendrin (8), p-hydroxybenzoic acid (9), (+)-catechin (10), gallic acid (11), nicotinamide (12), methyl gallate (13) and $1,2,3,4,6-penta-O-galloyl-{\beta}-D-glucose$ (14) by spectroscopic methods. Among these compounds, 1-3, 5, 6, 8 and 12 were isolated for the first time from this plant.

Effects of LY-117018 and Tamoxifen on Reproductive Organ in Rats (흰쥐에 있어서 LY-117018 및 Tamoxifen이 생식기관에 미치는 영향)

  • Park, Kyoung-Sik;Kwun, Jong-Kuk
    • Clinical and Experimental Reproductive Medicine
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    • v.14 no.1
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    • pp.61-70
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    • 1987
  • This study was carried out to investigate the effects of the antiestrogens, LY-117018 and tamoxifen on reproductive organ of ovariectomized immature rats and also to elucidate the mechanism of action of said compounds by bioassay. Each of LY-117018, tamoxifen and estradiol-17${\beta}$ was administered to ovariectomized immature rats at various dose levels. Forty hours after drug administration, tested rats were sacrificed and uterine wet weight, DNA and RNA contents in uterine and liver tissues were investigated. At the same time, uterine wet weight was also investigated with some other rats treated with 125${\mu}g$ of LY-117018 together with increasing doses of tamoxifen. Ovariectomized immature rats given 25${\mu}g$ single dose of each drug were sacrificed on Day 1, 2, 3, 4, and 5 after drug administration and uterine was weighed to estimate the duration of action of LY-117018 and tamoxifen. The results were summarized as follows: 1. The administration of LY-117018 or tamoxifen to ovariectomized rats increased uterine wet weight and DNA and RNA contents in uterine tissues with more increase in tamoxifen groups, but significant differences between groups treated at dose levels of 5${\beta}$ or more of both drugs were observed. Estradiol-17${\beta}$ groups showed significant increases in each group(P<0.01). 2. The administration of LY-117018 or tamoxifen to each group significantly increased DNA and RNA contents in liver tissues with more increase in tamoxifen groups. Estradiol-17${\beta}$ groups showed no significant differences between treatment groups of 5${\beta}$ or more. 3. Treatment with 125${\beta}$ of LY-117018 together with various doses of tamoxifen resulted in more increase of uterine wet weight than treatment with a single dose of LY-117018 or tamoxifen. 4. Treatment with 0.2${\beta}$ of LY-117018 or tamoxifen in ovariectomized rats decreased uterine wet weight,DNA and RNA contents in liver and uterine tissues compared with ovariectomized control. 5. The duration of effective action of LY-1l7018 and tamoxifen was 4 days or more. 6. There was significant difference(P<0.001) in uterine wet weight between Day 9after ovariectomy (two days after LY-117018 or tamoxifen treatment) and Day 10(63.7${\pm}$3.5mg, 39.2${\pm}$9.9mg, respectively).

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Kinetics of a Cloned Special Ginsenosidase Hydrolyzing 3-O-Glucoside of Multi-Protopanaxadiol-Type Ginsenosides, Named Ginsenosidase Type III

  • Jin, Xue-Feng;Yu, Hong-Shan;Wang, Dong-Ming;Liu, Ting-Qiang;Liu, Chun-Ying;An, Dong-Shan;Im, Wan-Taek;Kim, Song-Gun;Jin, Feng-Xie
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.343-351
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    • 2012
  • In this paper, the kinetics of a cloned special glucosidase, named ginsenosidase type III hydrolyzing 3-O-glucoside of multi-protopanaxadiol (PPD)-type ginsenosides, were investigated. The gene (bgpA) encoding this enzyme was cloned from a Terrabacter ginsenosidimutans strain and then expressed in E. coli cells. Ginsenosidase type III was able to hydrolyze 3-O-glucoside of multi-PPD-type ginsenosides. For instance, it was able to hydrolyze the 3-O-${\beta}$-D-(1${\rightarrow}$2)-glucopyranosyl of Rb1 to gypenoside XVII, and then to further hydrolyze the 3-O-${\beta}$-D-glucopyranosyl of gypenoside XVII to gypenoside LXXV. Similarly, the enzyme could hydrolyze the glucopyranosyls linked to the 3-O-position of Rb2, Rc, Rd, Rb3, and Rg3. With a larger enzyme reaction $K_m$ value, there was a slower enzyme reaction speed; and the larger the enzyme reaction $V_{max}$ value, the faster the enzyme reaction speed was. The $K_m$ values from small to large were 3.85 mM for Rc, 4.08 mM for Rb1, 8.85 mM for Rb3, 9.09 mM for Rb2, 9.70 mM for Rg3(S), 11.4 mM for Rd and 12.9 mM for F2; and $V_{max}$ value from large to small was 23.2 mM/h for Rc, 16.6 mM/h for Rb1, 14.6 mM/h for Rb3, 14.3 mM/h for Rb2, 1.81mM/h for Rg3(S), 1.40 mM/h for Rd, and 0.41 mM/h for F2. According to the $V_{max}$ and $K_m$ values of the ginsenosidase type III, the hydrolysis speed of these substrates by the enzyme was Rc>Rb1>Rb3>Rb2>Rg3(S)>Rd>F2 in order.

The Effects of Platelet- Derived Growth Factor-BB on the Expression of Bone Matrix Protein in the MC3T3-E1 Cells (MC3T3-E1 세포의 골기질 단백질 발현에 대한 혈소판유래성장인자-BB의 효과)

  • Kim, Myo-Sun;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.347-360
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    • 2000
  • Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of the polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. This process includes strictly regulated gene expression of several bone matrix proteins such as type I collagen and osteopontin, a 44 kDa phosphorylated glycoprotein, which has important roles in bone formation. The purpose of this study is to evaluate the effecs of PDGF-BB on the mRNA expression of bone matrix protein, type I collagen and osteopontin, in MC3T3- E1 cell culture. Cells were seeded at $5{\times}10^5$ cells in 10 ml of minimum essential medium alpha(${\alpha}-MEM$) containig 10% fetal bovine serum, 10 mM beta glycerophosphate. 0.1, 1, 10 ng/ml PDGF-BB were added to the cells for the day 3, 7, 14, 21, 28 and cultured for 24 hours. Type I collagen cDNA, Hf677, and osteopontin cDNA were used as probes for northern blot analysis. Total cellular RNA was purified at indicated day and northern blot analysis was performed. The results were as follows : Type I collagen mRNA expressions were higher at the day 3 and 7, and lower in the day 14, 21 in the control groups. In the experimental groups, mRNA expressions were increased when 0.1 ng/ml PDGF-BB were added on the day 3, 7, 21, and decreased in dose-dependent manner on the day 14, decreased at all added dose on the day 28. Osteopontin mRNA expressions were highest in the day 21 groups and lowest in the day 14 groups in the control groups. Interesting results were shown in the day 14 and 21 groups. We found that osteopontin mRNA level was increased in dose dependent manner in the day 14 groups, and decreased dose dependent manner in the day 21 groups. In conclusion, PDGF-BB may have various control effects on type I mRNA expression in the growth and differentiation process of MC3T3-E1 cells and may have contrary regulatory effects on osteopontin mRNA expression. For examples, when the baseline level of osteopontin mRNA was low, as in the day 14, PDGF-BB up-regulated osteopontin mRNA expression in dose dependent manner, and when the baseline level was high as in the day 21, PDGF-BB down-regulated dose dependent manner. Thus, it may be useful for clinical application in periodontal regeneration procedure if further study were performed.

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Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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Kinetics and Mechanism of Hydrolysis of Insecticidal Imidacloprid (살충성 Imidacloprid의 가수분해 반응 메카니즘)

  • Yu, Sung-Jae;Kang, Moon-Sung;Sung, Nack-Doo
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.53-57
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    • 1997
  • The rate of hydrolysis of insecticidal 1-(6-chloro-3-pyridylmethyl) -2-nitro-iminoimidazolidine (common name; imidacloprid) have been investigated in 15%(v/v) aqueous dioxane at $45^{\circ}C$. From the kinetics and non-kinetics data such as pH-effect, solvent effect(m=0.04, n=0.30 IT m<${\Delta}H^{\neq}=16.14kcal{\cdot}mol^{-1}\;&\;{\Delta}S^{\neq}=-0.03e.u.$), rate equation ($k_{obs.}=4.56{\times}10^{-3}[OH^-]$) and analysis of hydrolysis product, 1-(6-chloro-3-pyridylmethyl-2)-imidazolidinon, the hydrolysis mechanism of imidacloprid is proposed that the specific base catalyzed hydrolysis($K_{OH^-}$) through nucleophilic addition-elimination ($Ad_N-E$) mechanism proceed via intermediate, 1-(6-chloro-3- pyridylmethyl)-2-hydroxy-2-imidazolidinylisonitraminate (I) and ${\beta}$-3-(6-chloro-3-pyridylmethyl)aminoethyl-1-nitrourea(III). And the half-life(t1/2) of hydrolytic degradation at pH 8.0 and $45^{\circ}C$ was about 4.5 months.

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Chemical Components, Antioxidant Activity, and α-Glucoamylase Inhibitory Activity of a New Mushroom Variety 'Dahyang' (신품종 갈색양송이 '다향'의 화학성분과 항산화활성 및 α-Glucoamylase 저해활성)

  • Kim, Hong-Kyu;Yang, Euy-Seog;Park, Gi-Moon;Kim, Gwan-Hou;Kim, Hyun-Ho;Lee, Ka-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.8
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    • pp.1179-1183
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    • 2011
  • This study was conducted to investigate the quality characteristics, antioxidant activity, and ${\alpha}$-glucoamylase inhibitory activity of Dahyang, a Chungnam Agricultural Research & Extension Service's newly bred cultivar of brown button mushroom. Total phenolic compound contents of Dahyang and the no. 705 mushroom were 189${\pm}$12 mg% and 168${\pm}$8 mg%, respectively. The major free sugars in Dahyang were mannitol (3.11%), xylose (0.12%), and trehalose (0.08%). ${\beta}$-Glucan content was 28.34% in Dahyang and 26.55% in the no. 705 mushroom, respectively. Electron donating ability by DPPH in Dahyang and the no. 705 mushroom was 52.14% and 45.27% for the water extract, and 57.81% and 46.93% for the 80% ethanol extract, respectively. ${\alpha}$-Glucoamylase inhibitory activity in a 10 mg/mL concentration of water extract were was 33.25% in Dahyang and 29.22% in the no. 705 mushroom, respectively.