• Title/Summary/Keyword: AP-SF

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Antibiotic Resistance and R Plasmids in Edwardsiella tarda (양만장 사육조에서 분리한 Edwardsiella tarda의 약제 내성과 R Plasmid)

  • Choi, Min-Soon;Kim, Young-Gill
    • Journal of fish pathology
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    • v.7 no.1
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    • pp.37-46
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    • 1994
  • A total 103 strains of Edwardsiella tarda was isolated from eel culture-ponds and examined for drug resistance, distribution and transferabilities of R plasmids. The drugs used were lincomycin(LM), penicillin(PM), sulfamethazine(SF), sulfadimethoxine(SD), cephalosprin(CP), chloramphenicol(CH), streptomycin(SM), oxytetracycline(OT), ampicillin(AP), oxolinic acid(OA), kanamycin(KM), amikacin(AK), gentamicin(GM) and enrofloxacin(EF). Two strains were resistant to the all drugs used, and all isolates were multiply resistant to drugs(at least 8 among 14 drugs), mostly restricted to LM(103 strains), PM(103 strains), SD(103 strains), SF(103 strains), CP(102 strains), CM(101 strains), SM(100 strains), OT(94 strains), AM(92 strains), OA(80 strains), KM(60 strains), AK(30 strains), GM(19 strains) and EF(14 strains), in combination at high degree showing 34 different drug resistant patterns. The most frequently encountered patterns were LM PM SD SF CP CH SM OT AP OA KM(22 strains, 22.4%) followed by LM PM SD SF CP CH SM OT AP OT(12 strains, 11.7%). LM PM SD SF CP CH SM OT AP(7 strains, 6.8%), LM PM SD SF CP CH SM OT AP OA KM AK GM(6 strains, 5.8%) and LM PM SD SF CP CH SM OT AM OX KM AK GM(6 strains, 5.8%). Transfer experiment of drug resistance showed that of 103 resistant strains, 100 strains(97%) transferred part or all resistance to all drugs, indicating that most isolates carried conjugally transferrable R plasmids determining multiple drugs. The most frequently observed transferarble R plasmids were LM PM SD SF CP CH SM OT AP OA(10 strains), LM PM SD SF CP CH SM OT AP OX KM(7 strains) and LM PM SD SF CP CH AP OA(7 strains). These results sugested that eel culture-ponds were highly contaminated with different strains of Edwardsiella tarda, and that contaminated bacteria might be highly multiple resistant strains to drugs, carring transferable R plasmids.

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Anti-inflammatory activity of AP-SF, a ginsenoside-enriched fraction, from Korean ginseng

  • Baek, Kwang-Soo;Hong, Yong Deog;Kim, Yong;Sung, Nak Yoon;Yang, Sungjae;Lee, Kyoung Min;Park, Joo Yong;Park, Jun Seong;Rho, Ho Sik;Shin, Song Seok;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.39 no.2
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    • pp.155-161
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    • 2015
  • Background: Korean ginseng is an ethnopharmacologically valuable herbal plant with various biological properties including anticancer, antiatherosclerosis, antidiabetic, and anti-inflammatory activities. Since there is currently no drug or therapeutic remedy derived from Korean ginseng, we developed a ginsenoside-enriched fraction (AP-SF) for prevention of various inflammatory symptoms. Methods: The anti-inflammatory efficacy of AP-SF was tested under in vitro inflammatory conditions including nitric oxide (NO) production and inflammatory gene expression. The molecular events of inflammatory responses were explored by immunoblot analysis. Results: AP-SF led to a significant suppression of NO production compared with a conventional Korean ginseng saponin fraction, induced by both lipopolysaccharide and zymosan A. Interestingly, AP-SF strongly downregulated the mRNA levels of genes for inducible NO synthase, tumor necrosis factor-${\alpha}$, and cyclooxygenase) without affecting cell viability. In agreement with these observations, AP-SF blocked the nuclear translocation of c-Jun at 2 h and also reduced phosphorylation of p38, c-Jun N-terminal kinase, and TAK-1, all of which are important for c-Jun translocation. Conclusion: Our results suggest that AP-SF inhibits activation of c-Jun-dependent inflammatory events. Thus, AP-SF may be useful as a novel anti-inflammatory remedy.

Polyhedra Productions of Recombinant Autographa californica Nucle- opolyhedroyiruses Containing Additional Polyhedrin of Autographa Cali- fornica, Bombyx mori or Spodoptera exigua Nucleopolyhedrovirus

  • Chang, Jin-Hee;Roh, Jong-Yul;Jin, Byung-Rae;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.13-18
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    • 2001
  • The role of polyhedrin in the polyhedra production in baculovirus Autograha californica Nucelopolyhedro-sisvirus (AcNPV) was studied by over-expression of AcNPV polyhedrin or heterologous polyhedrin from Bombyx mori (Bm) NPV or Spodoptera exigua (Se) NPV. The transfer vectors containing additional polyhedrin from AcNPV, BmNPV, or SeNPV were constructed and cotransfected with bacmid bApGOZA into Sf9 cells. The resulting recombinants, designated as vApAcPol, vApBmPol, and vApSePol were tonstructed, and the polyhedra production of the recombinant was characterized. All of the recombinants produced polyhedra in the nucleus, and the polyhedrin was over-expressed. Among three recombinants, vApAcPol and vApBmPol were discriminated by their larger polyhedra size than that of wild type AcNPV, and vApSePol also produced larger polyhedra than wild type SeNPV polyhedra.

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A Comparative Study on the Changes of Pelvic Alignment between AP View of the Pelvis in Standing and Supine Position (Pelvic AP X-ray 촬영 자세에 따른 골반변위 변화의 비교 연구)

  • Lee, Kyung-Moo;Park, Dong-Su;Kim, Soon-Joong;Jeong, Su-Hyeon
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.4
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    • pp.161-169
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    • 2008
  • Objectives : This study was carried out to investigate the relationship of pelvic alignment between AP view of the pelvis in standing and supine position. Methods : Sixteen healthy peoples who get $51.59{\pm}4.14$ as average in SF-36 were evaluated by X-ray findings. After measuring innomiate measurement, off centering measurement, sacral ala measurement, illium shadow measurement, the area of obturator foramen, those were analyzed statistically. Results : It was not all to be corresponded to distort pelvic alignment of AP view of the pelvis in standing and in supine. Sometimes it was the opposite result. Conclusions : These results suggest that the diagnosis of pelvic alignment to go through on each position is brought about disagreement with each other.

Development of Pre-Mix Cement for 150 MPa Ultra High Strength Concrete (설계강도 150 MPa 초고강도 콘크리트용 시멘트 결합재의 개발)

  • Hwang, Yin-Soong;Kim, Seong-Su;Cha, Wan-Ho;Kwon, O-Bong;Sohn, Yu-Shin;Lee, Seung-Hoon
    • Proceedings of the Korea Concrete Institute Conference
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    • 2006.05b
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    • pp.25-28
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    • 2006
  • This study investigated pre-mixed cement combined with ordinary portland cement, BF and SF, in order to manufacture cement binder, which is possible to produce 150MPa ultra high strength concrete. The BF used in this study reduces and control hydration heat. It can also improve concrete fluidity, while AP increases hydration product and accelerates reaction of BF. SF has micro filler effect and makes pozzolanic reaction. It also fabricates high density internal organization. This developed pre-mixed cement can reduce hydration heat and increase hydration product. It is possible to fabricate high density organization and to secure homogeneity. The mock-up test of ultra high strength concrete showed excellent dispersibility and workability and indicated compressive strength more than 150MPa at 28 days.

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High-Level Expression of T4 Endonuclease V in Insect Cells as Biologically Active Form

  • Kang, Chang-Soo;Son, Seung-Yeol;Bang, In-Seok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1583-1590
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    • 2006
  • T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.

Optimal Conditions for the Expression of Glycoprotein E2 of Classical Swine Fever Virus using Baculovirus in Insect Cells

  • Bae, Sung Min;Lee, Seung Hee;Kwak, Won Suk;Ahn, Yong Oh;Shin, Tae Young;Woo, Soo Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.207-213
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    • 2014
  • The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein $E^{rns}$ (E0), E1 and E2. Among them, E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. In this study, to determine the optimal expression conditions of glycoprotein E2 using baculovirus system, we investigated the influence of insect cells and media to the expression of recombinant E2. Recombinant virus containing glycoprotein E2 coding gene was constructed with bApGOZA DNA. Expression of the glycoprotein E2 was analyzed by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies. Expression of glycoprotein E2 in Sf21 cells was first observed after 3 days and reached a maximum on the 5th day after infection. Furthermore, the highest levels of glycoprotein E2 expression were observed at multiplicity of infection (MOI) of 5. When three different insect cell lines (Sf21, High-Five and Se301) were tested, High-Five cells showed the highest production. In addition, four different serum-free and serum-supplemented media, respectively, were tested for the expression of glycoprotein E2 and the budded virus (BV) titers. As a result, serum-supplemented medium provided the best conditions for protein production and the BV yield.

Extraction Characteristics of Flavonoids from Lonicera flos by Supercritical Fluid Carbon Dioxide ($SF-CO_2$) with Co-solvent (초임계유체 $CO_2$ 및 Co-solvent 첨가에 따른 금은화(Lonicera fles)의 Flavonoid류 추출특성)

  • Suh, Sang-Chul;Cho, Sung-Gill;Hong, Joo-Heon;Choi, Yong-Hee
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.183-188
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    • 2005
  • Effects of co-solvent polarity, citric acid, pressure, temperature, run time, and co-solvent ratio on extraction of major flavonoids from Lonicera Flos were investigated using supercritical fluid $CO_{2}(SF-CO_{2})$. HPLC analysis revealed addition of pure methanol resulted in low extraction yield of major flavonoids, luteoloin (Lu), Quercetin (Qu), Apigenin (Ap). Under same condition, as co-solvent polarity increased, yields of major flavonoids increased gradually, At optimum co-solvent extraction condirion of 60% aqueous methanol (10%, v/v), yields of Lu, Qu, and Ap were 42.09, 28.18, and 3.49 mg/100 g, respectively. Addition of citric acid to 60% aqueous methanol gave higher, with addition of 1% citrie acid resulting in highest yields of 63.2 (Lu), 39.35 (Qu), and 5.79 (Ap) mg/100 g. Optimum extraction conditions of major flavonoids were 200 bar, $50^{\circ}C$, 60 min, and $CO_{2}$-methanol-water(20: 1.8: 1.2).

Nucleotide Sequence and Cloning of sfs4, One of the Genes Involved in the CRP-Dependent Expression of E. coli mal Genes. (CRP 의존성 maltose 대사 촉진 유전자 sfs4의 클로닝 및 염기배열 결정)

  • Chung, Soo-Yeol;Cho, Moo-Je;Jeong, Hee-Tae;Choi, Yong-Lark
    • Applied Biological Chemistry
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    • v.38 no.2
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    • pp.111-117
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    • 1995
  • In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

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