• Title/Summary/Keyword: PHA-P

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Evaluation of Vi-Passive Hemagglutination, SD$^{(R)}$ Kit, and Widal Test for Serological Diagnosis of Typhoid Fever (장티푸스의 혈청학적 진단을 위한 Vi-수동혈구응집법, SD$^{(R)}$ Kit 및 Widal 시험에 대한 효용성 평가)

  • Kim, Sung-Hun;Kim, Shuk-Ho;Lee, Deog-Yong;Lee, Esther;Park, Mi-Sun;Lee, Bok-Kwon
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.219-222
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    • 2010
  • In this study, we evaluated Vi-passive hemagglutination (Vi-PHA), SD Salmonella Typhi IgG/IgM ($SD^{(R)}$ kit) and Widal test for the rapid laboratory diagnosis of typhoid fever patients. A total of 36 serum samples from febrile patients in Korea from 2005 to 2006 were used. Among 36 patients, 27 were fever patients without typhoid, 9 were typhoid fever. Vi-PHA showed 8 positive results out of 9 typhoid fever patients (sensitivity 88.9%) and 1 positive and 26 negative results out of 27 febrile patients without typhoid (specificity 96.3%). The sensitivity and the specificity of $SD^{(R)}$ kit were 100% and 92.6%, respectively. However, the sensitivity and the specificity of Widal O & H tests were 88.9%, 100%, and 77.8%, 70.4%, respectively. Consequently, Widal H and $SD^{(R)}$ kit showed higher sensitivity and Vi-PHA showed higher specificity. To efficient diagnosis, Vi-PHA may be sufficient diagnosis method in acute cases and $SD^{(R)}$ kit and Widal test may be sufficient in sporadic area and high risk group.

Effects of Glucose and Acrylic acid Addition on the Biosynthesis of Medium-Chain-Length Polyhydroxyalkanoates by Pseudomonas chlororaphis HS21 from Plant Oils (Pseudomonas chlororaphis HS21에 의한 식물유로부터 Medium-Chain-Length Polyhydroxyalkanoates 생합성이 미치는 포도당 및 아크릴산의 첨가 효과)

  • Chung Moon-Gyu;Yun Hye Sun;Kim Hyung Woo;Nam Jin Sik;Chung Chung Wook;Rhee Young Ha
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.225-231
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    • 2005
  • The characteristics of cell growth and medium-chain-length polyhydroxyalkanoate (MCL-PHA) biosynthesis of Pseudomonas chlororaphis HS21 were investigated using plant oils as the carbon substrate. The organism was efficiently capable of utilizing plant oils, such as palm oil, corn oil, and sunflower oil, as the sole carbon source for growth and MCL-PHA production. When palm oil (5 g/L) was used as the carbon source, the cell growth and MCL-PHA accumulation of this organism occurred simultaneously, and a high dry cell weight (2.4 g/L) and MCL-PHA ($40.2\;mol{\%}$ of dry cell weight) was achieved after 30 hr of batch-fermentation. The repeating unit in the MCL-PHA produced from palm oil composed of 3-hydroxyhexanoate ($7.0\;mol{\%}$), 3-hydroxyoctanoate ($45.3\;mol{\%}$), 3-hydroxydecanoate ($39.0\;mol{\%}$), 3-hydroxydodecanoate ($6.8\;mol{\%}$), and 3-hydroxytetradecanoate ($1.9\;mol{\%}$), as determined by GC/MS. Even though glucose was a carbon substrate that support cell growth but not PHA production, the conversion rate of palm oil to PHA was significantly increased when glucose was fed as a cosubstrate, suggesting that bioconversion of some functionalized carbon substrates to related polymers in P chlororaphis HS21 could be enhanced by the co-feed of good carbon substrates for cell growth. In addition, the change of compositions of repeating units in MCL-PHAs synthesized from the plant oils was markedly affected by the supplementation of acrylic acid, an inhibitor of fatty acid ${\beta}-oxidation$. The addition of acrylic acid resulted in the increase of longer chain-length repeating units, such as 3-hydroxydodecanoate and 3-hydroxytetradecanoate, in the MCL-PHAs produced. Particularly, MCI-PHAs containing high amounts of unsaturated repeating units could be produced when sunflower oil and corn oil were used as the carbon substrate. These results suggested that the alteration of PHA synthesis pathway by acrylic acid addition can offer the opportunity to design new functional MCL-PHAs and other unusual polyesters that have unique physico-chemical properties.

Blastogenesis of splenic Iymphocytes to specific antigens and PHA in Paragonianus westermani infected mice (폐흡충 항원 및 Phytohemagglutinin에 의한 마우스 비장 림프구의 아세포화(Blastogenesis)반응)

  • Min, Duk-Young;Shin, Myeong-Heon;Choi, Ryung
    • Parasites, Hosts and Diseases
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    • v.30 no.1
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    • pp.43-48
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    • 1992
  • Paragonimus westermani is a common fluke in Uorea. The present study aimed to observe the cell mediated immune response in experimental paragonimiasis of mice. The mouse (BALB/c) was orally inoculated with 40 metacercariae of P. westermani from Cambaroides similis. During the infection (1, 2, 4, 6 weeks) of mouse, blastogenic response of splenic Iymphocytes to P. westermani adult antigen, metacercaria antigen, and PHA were observed. Sera from infected and noninfected mice added to normal mouse splenic Lymphocytes with or without PHA. The blastogenic response of splenic Lymphocytes to PHA was reduced after 1 week of infection. However after 6 weeks of infection, the response was restored to the control level. The blastogenic response of splenic Iymphocytes to P. westermani adult or metacercaria antigen increased significantly on 1 week after infection, and maintained up to 6 weeks after infection. The response of non-infected mice was suppressed by addition of the infected mouse serum. The present results suggested that cellular immunity was involved in P. westermani infected mice and that P. westermani anti.serum inhibited proliferation of T Iymphocytes.

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The Studies of Copolymer, Poly (3-hydroxybutyrate-co-3-hydroxy valerate) Synthesis in Recombinant pha+ Escherichia coli

  • Lee, Ha-Young;Rhie, Ho-Gun
    • Proceedings of the Zoological Society Korea Conference
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    • 1996.10a
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    • pp.195.2-195
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    • 1996
  • acs gene cloning was constructed by subcloning the 2.2-kb MunI-MunI restriction fragment of 638 and 639 which include acs gene from the kohara phage into the unique EcoRI site of pUC18 and pJM9131 containing the PHA biosynthesis genes. Then recombinant E. coli fadRatoC(Con) mutants containing the polyhydroxyalkanoate(PHA) biosynthesis genes are able to incoporate s significant levels of 3-hydroxyvalerate (3HV) into the copolymer [P(3HB-co-3HV)]. Quantitative determination of PHB and P(3HB-co-3HV) was performed by gas-chromatographic analysis of extracts obtained from methanolysis of lyophilized cells.

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Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli from Sucrose (재조합 대장균에서 수크로즈로부터의 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구)

  • Oh, Young Hoon;Kang, Kyoung-Hee;Shin, Jihoon;Song, Bong Keun;Lee, Seung Hwan;Lee, Sang Yup;Park, Si Jae
    • KSBB Journal
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    • v.29 no.6
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    • pp.443-447
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    • 2014
  • Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) and engineered C. propionicum propionyl-CoA transferase ($Pct_{Cp}$), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16 mol%LA) synthesized in E. coli W using sucrose as carbon source were $1.53{\times}10^4$ ($M_n$) and $2.78{\times}10^4$ ($M_w$), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.

Inhibition of Intimal Hyperplasia by Perivascular Delivery of Paclitaxel Using Poly(n-butylmethacrylate) or Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) in Balloon-Injured Rat Carotid Arteries (쥐 경동맥 손상 모델에서 Poly(n-butylmethacrylate) 혹은 Poly(3-hydroxybutyrate-co-4-hydroxybutyrate)를 이용한 Paclitaxel 국소 전달 요법의 신생내막 형성 억제 효과)

  • Kwon, Jin-Sook;Lee, Ho-Yeon;Heo, Shin-Haeng;Park, Rho-Kwan;Shim, Tae-Jin;Kim, Yu-Kyug;Kim, Beom-Soo;Kim, Dong-Woon
    • Polymer(Korea)
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    • v.32 no.3
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    • pp.284-289
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    • 2008
  • Polymer is a critical component of local drug delivery to prevent restenosis. This study tested whether poly(n-butylmethacrylate)(PBMA) and poly(3-hydroxybutyrate-co-4-hydroxybutyrate)(PHA) was candidates for this purpose. In vitro release of paclitaxel from PBMA and PHA loaded with 10% paclitaxel exhibited a triphasic release profile, with a fast initial and intermediate second phase followed by a slow release phase. Perivascular delivery of paclitaxel using these films inhibited neointimal hyperplasia in balloon-injured rat carotid arteries. The paclitaxel-loaded PBMA or PHA groups showed significant neointimal formation reductions versus the control groups (PBMA vs control: $0.03{\pm}0.02$ vs $0.10{\pm}0.01\;mm^2$, p<0.05; PHA vs control: $0.04{\pm}0.03$ vs $0.09{\pm}0.01\;mm^2$, p<0.05). This study suggests that PBMA and PHA could be good candidate polymers of local drug delivery to prevent restenosis. Perivascular delivery using these films represents a possible approach for prevention of restenosis. These can be candidate polymers for drug eluting stents.

Synthesis of Poly[3-hydroxybutyrate-co-3-hydroxyvalerate] by Recombinant Escherichia coli from Whey (재조합 대장균에 의한 유청으로부터 Poly[3-hydroxybutyrate-co-3-hydroxyvalerate] 합성)

  • 김범수;이상엽
    • KSBB Journal
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    • v.18 no.5
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    • pp.404-407
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    • 2003
  • Two recombinant Escherichia coli strains, GCSC6576 harboring a plasmid pSYL107 containing the Ralstonia eutropha polyhydroxyalkanoate (PHA) biosynthesis genes and a fadR atoC mutant LS5218 harboring a plasmid pJC4 containing the Alcaligenes latus PHA biosynthesis genes were compared for their ability to synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV) from whey. The 3HV fraction could be increased by acetic acid induction and oleic acid supplementation in flask cultures of recombinant E. coli GCSC6576. With the pH-stat fed-batch culture of recombinant E. coli LS5218, we obtained a cell concentration, a P(3HB-co-3HV) concentration, a P(3HB-co-3HV) content, and a 3HV fraction of 31.8 g/L, 10.6 g/L, 33.4%, and 6.26 mol%, respectively in 39 h.

Synthesis of Copolymeric PHA by Hydrogenophaga pseudoflava and Ralstonia eutropha H16 from Vari-ous Lactones and Their Microstructural Studies (락톤류로부터 Hydrogenophaga pseudoflava와 Ralstonia eutropha H16 두 세균에 의한 공중합 PHA의 합성 및 미세구조적 특성 연구)

  • Jang, Young-Ok;Nam, Won;Choi, Mun-Hwan;Song, Jae-Jun;Yoon, Sung-Chul
    • Microbiology and Biotechnology Letters
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    • v.28 no.2
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    • pp.71-79
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    • 2000
  • Two typess of copolyesters, poly(3-hydroxybutyric acid-co-4-hydroxy-butyric acid)[P(3HB-co-4HB] and poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid)[P(3HB-co-3HV)], with various monomer ratios and different degree of microstructural heterogeneity were synthesized from Ralstonia eutropha H16 and Hydrogenophaga pseudoflava by using ${\gamma}$-butyrolactone and ${\gamma}$-valerolactone, respectively. The two bacteria showed a large difference in the utilization of ${\gamma}$-butyrolactone for cell growth and PHA synthesis. H. pseudoflava synthesized P(3HB-co-4HB) copolyesters with a wide range of 4HB content from 13 to 96 mol% depending on culture conditions, whiel R. eutropha H16 was able to synthesize the copolyesters containing less than 20 mol% of 4HB. An increase in the 4HB content in the P(3HB-co-4HB) copolyesters synthesized by H. pseud-oflava induced an lowering of their melting temperatures as well as their enthalpies of fusion. The increase in the 4HB content, however, increased the rate of degradation by an extracellular P(3HB) depolymerase. NMR spectros-copy and differential scanning calorimetry showed that the P(3HB-co-4HB) copolyesters from H. pseudoflava were generally microstructurally heterogeneous. The P(3HB-co-4HB) copolyesters) synthesized by R. eutropha H16 were rather random copolymers showing less microstructural heterogeneity than those synthesized by H. pseudoflava. The NMR D value analysis suggested that the monomer distribution of the P(3HB-co-3HV) copolymers from the two bacteria were relatively random.

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Vaccination Studies against Fowl Pox in Chickens (닭에서 계두 예방 접종에 관한 연구)

  • Shil, N.K.;Rahman, M.S.;Paul, S.;Cha, S.Y.;Jang, H.K.;Song, H.J.
    • Korean Journal of Poultry Science
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    • v.34 no.4
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    • pp.253-257
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    • 2007
  • Fayoumi chicks were vaccinated with fowl pox virus vaccine and pigeon pox virus vaccine. The protective potentiality of the two vaccines was compared in field condition in Bangladesh. The percentage of 'take reaction' was assessed to conclude its relationship with better immune response and recorded 93.33% and 100% in birds of group B and group C, respectively. The mean passive hemagglutination (PHA) antibody titre after primary vaccination was $32{\pm}14.81$ in group B and $33{\pm}13.66$ in group C. Following booster vaccination, the mean PHA titres level at pre challenge of group B was $46.93{\pm}16.52\;and\;55.46{\pm}14.64$ in group C. The PHA titre of group B and C at two weeks post challenge recorded $93.86{\pm}33.04\;and\;110.93{\pm}29.29$, respectively. PHA titre significantly (P<0.01) increased after vaccination and post challenge compared to pre- vaccination titre. There was significant variation (p<0.01) of PHA titre at pre challenge in these groups using different vaccine combinations, but all the vaccinated birds resisted challenge infection.

Isolation of Pseudomonas putida BM01 Accumulating High Amount of $PHA_{MCL}$

  • Song, Jae-Jun;Yoon, Sung-Chul
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.126-133
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    • 1994
  • A Pseudomonas putida strain able to accumulate high amount of polyesters of medium-chain-length 3-hydroxyalkanoic acids ($PHA_{MCL)$) was isolated from soil in a landfill site using an enrichment technique. Culture condition of the isolated strain for polyester production in a one-step culture was optimized in a mineral-salts medium against pH and concentrations of ammonium sulfate, carbon source(e.g., octanoate), and phosphate. The optimal values for maximal cell growth and PHA accumulation were: pH; 7$\sim$8, $(NH_4)_2SO_4$; 8 mM, octanoate; 40 mM. The optimum temperature was in the range of $20\sim30^{\circ}C$, which was rather broader than in other bacteria. Cell growth was strongly inhibited by the phosphate limitation to less than 1 mM. An increase of phosphate concentration above 1 mM showed little effect on cell growth and polyester accumulation. When the strain was grown on octanoate under this optimized condition it produced 3.4 g dry biomass per liter and yielded 1.7 g PHA per liter amounting to 53 wt% of dry cells. The monomer units composing the polyester synthesized from octanoate were 3-hydroxyoctanoate (3HO), 3-hydroxycaproate (3HC), and 3-hydroxybutyrate (3HB) (85:13:2, mole ratio). Other low linear $C_3\simC_{10}$ monocarboxylic acids were also tested for polyester production.

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