• 제목/요약/키워드: P.E.B system

검색결과 414건 처리시간 0.025초

Fumonisin B1에 의한 세균바이러스 DNA손상 (Bacterial Virus DNA Damage Caused by Fumonisin B1)

  • 이길수;조성국
    • 한국환경성돌연변이발암원학회지
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    • 제19권1호
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    • pp.34-38
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    • 1999
  • Fumonisin B1 is a secondary metabolite of Fusarium moniliforme, a contaminant of corn and corn product. Fumonisin B1 has been shown to be responsible for major toxicological effects of the fungus in rats, horses, and pigs. Fumonisin B1 induced λ DNA fragmentation, which was increased with incubation time, reducing agent NADPH and metal ion (Cu2+). The DNA damage was inhibited by dimethyl sulfoxide (DMSO) or mannitol as radical scavenger for free radicals. DNA fragmentation, induced by fumonisin B1 in the presence of 1 mM NADPH and 0.1 mM CuCl2, was inhibited by 100 mM DMSO. By the in vitro reaction of fumonisin B1 with supercoiled plasmid pBR322 DNA, plasmid DNA was relaxed, eventually linearized in the agarose gel electrphoresis. From rifampicin sensitive E. coli CSH138 in bacterial mutagenesis system, the rifampicin resistant E. coli mutants were obtained by fumonisin B1. These results suggest that fumonisin B1 may be a possible environmental mutagen in bacterial mutagen assay system.

Immunization with a Genetically Engineered Uropathogenic Escherichia coli Adhesin-Escherichia coli Enterotoxin Subunit A2B Chimeric Protein

  • Lee, Yong-Hwa;Kim, Byung-O;Pyo, Suhk-Neung
    • Biomolecules & Therapeutics
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    • 제13권2호
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    • pp.101-106
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    • 2005
  • The generation of secretory IgA antibodies (Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the heat-labile Escherichia coli enterotoxin A2B (ltxa2b) gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimH/ltxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and westem blotting using antibodies to the maltose binding protein (MBP) or the heat labile E. coli subunit B (LTXB), plus the N-terminal amino acid sequence was analyzedd. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/LTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of LTXB. thisstudy also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA (sIgA) and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked LTXA2B acts as a useful mucosal adjuvant, and that adhesin/LTXA2A chimeric protein might be a potential antigen for oral immunization against UPEC.

테스토스테론 물질 검출을 위한 in vitro TCD 시스템 구축 (Establishment of an In Vitro TCD (Testosterone Compound Detection) System)

  • 이동근;조정권;이상현
    • 생명과학회지
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    • 제29권10호
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    • pp.1159-1163
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    • 2019
  • 남성호르몬 감소와 관련된 남성갱년기에 대한 관심이 고조되고 있지만, 남성호르몬의 정량을 위해 항체를 이용하는 고가의 kit가 이용되고 있다. 본 연구에서는 in vitro 전사 활성 시험법을 이용하여 남성 스테로이드호르몬의 활성 혹은 농도를 검증하는 시스템을 구축하였다. 테스토스테론-AR (androgen receptor) 복합체와 반응하는 ARE-AdE1bTATA 염기서열이 삽입되고 리포터로 luciferase를 발현하는 테스토스테론 유사활성 검증 리포터 플라스미드인 pGL2-Neo-ARE-AdE1BTATA를 제조하고, 인체 전립선암 세포인 LNcap-LN3 세포에 stable transfection을 실시하였다. 구축된 LNcap-LN3/pGL2-Neo-ARE-AdE1BTATA TCD (testosterone compound detection) 시스템은 표준물질인 테스토스테론의 $10^{-13}{\sim}10^{-8}M$ 범위에서 농도 증가에 비례하는 정량성을 보였다. 이 연구에서 확립된 in vitro TCD 시스템을 이용하면 천연물 유래 테스토스테론 유사물질 및 테스토스테론 저하물질의 대량 탐색 등이 가능할 것이므로, 건강기능성 식품이나 의약품 신소재의 개발에 기여할 것이다.

Application of Pulsed Electric Fields with Square Wave Pulse to Milk Inoculated with E. coli, P. fluorescens, and B. stearothermophilus

  • Shin, Jung-Kue;Jung, Kwan-Jae;Pyun, Yu-Ryang;Chun, Myong-Soo
    • Food Science and Biotechnology
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    • 제16권6호
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    • pp.1082-1084
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    • 2007
  • Ultra-high temperature (UHT) processed full fat milk inoculated with Escherichia coli, Pseudomonas fluorescens, and Bacillus stearothermophilus was exposed to 30-60 kV/cm square wave pulsed electric field (PEF) with $1\;{\mu}sec$ pulse width, and $26-210\;{\mu}sec$ treatment time in a continuous PEF treatment system. Eight log reduction was obtained for E. coli and P. fluorescens and 3 logs reduced for B. stearothermophilus under PEF treatment conditions of $210\;{\mu}sec$ treatment time, 60 kV/cm pulse intensity at $50^{\circ}$. There was no significant change in pH and titration acidity of milk after PEF treatment. The electrical energy required to achieve 8 log reduction for E. coli and P. fluorescens was estimated to be about 0.74 kJ/L.

Induction of a systemic IgG and secretory IgA responses in mice by peroral immunization with uropathogenic Escherichia coli adhesin protein coupled to cholera toxin A2B subunits

  • Lee, Yong-Hwa;Kim, Byung-Oh;Rhee, Dong-Kwon;Pyo, Suh-Kneung
    • Biomolecules & Therapeutics
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    • 제11권3호
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    • pp.157-162
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    • 2003
  • The generation of secretory IgA antibodies(Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimHIctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was analyzed. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/CTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of CTXB. This study also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin/CTXA2B chimeric protein might be a potential antigen for oral immunization against UPEC.

Comparative Study on Characterization of Recombinant B Subunit of E. coli Heat-Labile Enterotoxin (rLTB) Prepared from E. coli and P. pastoris

  • Ma, Xingyuan;Yao, Bi;Zheng, Wenyun;Li, Linfeng
    • Journal of Microbiology and Biotechnology
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    • 제20권3호
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    • pp.550-557
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    • 2010
  • Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6$\times$His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at $30^{\circ}C$, whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at $37^{\circ}C$. The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6$\times$His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6$\times$His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.

二中心 H${\u}$ckel 法의 應用 脂肪族 아민의 염기도 [Ⅰ] (Application of Two Centre HTEX>$\''{u}$ckel Method for Basicity of the Aliphatic Amines)

  • 박병각;이무상;홍영석
    • 대한화학회지
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    • 제15권3호
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    • pp.147-152
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    • 1971
  • On the baisis of the calculations by the two centre Huckel method for sigma electron system, the electronic structure of the various aliphatic amines including ammonia, and the relationship between the observed $pK_b$ and the change in the sigma electronic energy, ${\Delta}E{\sigma}$ in the course of protonation are discussed. A parallelism is observed between the $pK_b$ and the calculated ${\Delta}E{\sigma}$ of the amines. Also, it is observed that the electron densities of hydrogen atom directly bonded to nitrogen of the amines, likewise have a linear relationship with the $pK_b$. Therefore, the basicity of the aliphatic amines may be estimated qualitatively by means of the two center Huckel method.

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분석기법의 차이에 따른 현장시료의 VOC 분석결과 비교연구: 분석오차의 발생 양상과 원인 (A comparative analysis of volatile organic compound levels in field samples between different gas chromatographic approaches)

  • 안지원;수딜 팬디;김기현
    • 분석과학
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    • 제23권5호
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    • pp.465-476
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    • 2010
  • 본 연구에서는 대기환경시료 및 현장조건에서 채취한 현장시료들 중, 악취성분 및 주요 VOC 성분들 중에서 여러 유형의 관리대상으로 지정 중인 benzene, toluene, p-xylene, styrene, methyl ethyl ketone의 농도분석 방식들을 비교하였다. 이때, GC/FID와 튜브를 연계한 F-T 분석방식과 GC/MS와 백(bag)을 연계한 M-B방식으로 같은 시료를 각각 분석하고 이들 결과 값을 비교하였다. 상관분석결과, toluene과 pxylene에서 F-T와 M-B방법간의 결과에서 큰 상관성을 보이는 것으로 나타났다(P < 1.4E-08). 양 기기간의 오차를 percent difference (PD)와 같이 산술적으로 구하여 평가한 결과, benzene과 toluene의 농도는 상대적으로 비슷한 경향을 보였다. 그러나 분석대상 성분들 간에 양 방식의 차이는 평균적으로 35%가 넘으면서 F-T와 M-B간의 오차가 일정 수준이상으로 존재하였다. 그리고 F-T/M-B ratio가 대부분 1 이하로 나타나면서, M-B의 결과 값이 F-T에 비해 상대적으로 크게 나타나는 경향을 보였다. 양 분석기기로부터 산출한 결과 값을 통계적으로 판단하기 위해, t-test를 실시하였다. 그 결과, 95% 신뢰수준에서 toluene, pxylene, styrene, methyl ethyl ketone 성분에서는 두 시스템간의 결과 값에 차이가 유의한 것으로 나타났다(P < 0.043). 그러나 전반적으로 양 분석기법의 차이에 따른 오차는 상당 부분 선택한 표준시료의 성상 및 백내부의 시료손실 등과 같은 요인에 크게 영향을 받는 것으로 나타났다.

다양한 IPS Empress Esthetic$^{(R)}$ ingot으로 제작한 laminate veneer의 최종 색조에 대한 영향 (Influencing factors on the final color of laminate veneer restorations with various IPS Empress Esthetic$^{(R)}$ ingots)

  • 양미선;김석규
    • 대한치과보철학회지
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    • 제49권4호
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    • pp.308-315
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    • 2011
  • 연구 목적: IPS Empress Esthetic$^{(R)}$ system의 ingot은 다양한 반투명도를 가지고 있지만, 실제 다양한 색조의 자연치를 배경으로 각각의 ingot을 laminate veneer로 제작했을 때 어느 정도의 차폐능력을 나타내는지 알 수 없다. 본 연구의 목적은 반투명도가 서로 다른 ingot으로 제작한 IPS Empress Esthetic$^{(R)}$ laminate veneer 수복 시 다른 색상의 지대치상에서 최종 색조의 차이(color difference, ${\Delta}E$)를측정, 분석해서 그 차폐효과를 평가하고, 최종 색조에 미치는 영향을 알아보고자 하였다. 연구 재료 및 방법: Laminate veneer 수복을 위해 삭제된 6가지 다른 색조(A1, A3, A4, B2, B3, C3)의 자연치아 위에 반투명도가 서로 다른 6가지 IPS Empress Esthetic$^{(R)}$ ingot (E 01, E 03, E OC1, E TC1, E TC2, E TC3)으로 제작된 총 두께 0.6 mm의 도재 시편들을 translucent shade의 시적용 합착제로 시적한 뒤 각 시편 조합의 색조를 색채계(colorimeter)를 이용하여 측정하였다. 서로 다른 색조의 지대치아 위에서 측정된 도재시편들의 색차를 비교, 분석하였다. 결과: 지대치 색조 및 coping의 반투명도에 따라서 laminate veneer 수복물의 최종 색조가 다르게 나타났다(P<.05). 모든 도재 시편에서 A3, B3, 그리고 C3 shade를 가진 지대치들 사이의 평균 색차값(${\Delta}E$)이 2.7 이하로 나타났고, 지대치들 간의 색조가 A3 와 A4, B3 와 A4, 그리고 C3 와 A4 인 경우 평균 색차값은 몇 가지 도재 시편에서 2.7 이하를 나타내어 laminate veneer 수복물에 의한 지대치 색조의 차폐효과를 보여 주었다. 반면, A1 과 B2 shade의 지대치는 다른 지대치들과 비교하여 높은 색조 차이를 보였으며, 특히A4와 B2, A3 와 B2, 그리고 A1 과 A4 색조의 지대치들 사이에는 그 색조 차이가 컸다. 결론: IPS Empress Esthetic$^{(R)}$ coping으로 제작된 laminate veneer의 최종 색조는 지대치 색조 및 coping의 반투명도에 의해 영향을 받았다. 서로 다른 $a^*$$b^*$ 값을 나타내는 지대치들의 색상 차이는 어느 정도 차폐 되는 효과를 보여주고 있으나, 명도($L^*$)의 차이가 많이 나타나는 지대치 색조들을 차폐하는 것은 한계가 있었다.

The effect of ceramic thickness and number of firings on the color of a zirconium oxide based all ceramic system fabricated using CAD/CAM technology

  • Bachhav, Vinay Chila;Aras, Meena Ajay
    • The Journal of Advanced Prosthodontics
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    • 제3권2호
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    • pp.57-62
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    • 2011
  • MATERIALS AND METHODS. Ceramics have a long history in fixed prosthodontics for achieving optimal esthetics and various materials have been used to improve ceramic core strength. However, there is a lack of information on how color is affected by fabrication procedure. The purpose of this study was to evaluate the effects of various dentin ceramic thicknesses and repeated firings on the color of zirconium oxide all-ceramic system (LavaTM) fabricated using CAD/CAM technology. MATERIALS AND METHODS. Thirty disc-shaped cores, 12 mm in diameter with a 1 mm thickness were fabricated from zirconium oxide based all ceramic systems ($Lava^{TM}$, 3M ESPE, St Paul, MN, USA) and divided into three groups (n = 10) according to veneering with dentin ceramic thicknesses: as 0.5, 1, or 1.5 mm. Repeated firings (3, 5, 7, or 9) were performed, and the color of the specimens was compared with the color after the initial firing. Color differences among ceramic specimens were measured using a spectrophotometer (VITA Easyshade, VITA Zahnfabrik, Bad $S{\ddot{a}}ckingen$, Germany) and data were expressed in CIELAB system coordinates. A repeated measures ANOVA and Bonferroni post hoc test were used to analyze the data (n = 10, ${\alpha}=.05$). RESULTS. $L^{\ast}a^{\ast}b^{\ast}$ values of the ceramic systems were affected by the number of firings (3, 5, 7, or 9 firings) (P<.001) and ceramic thickness (0.5, 1, or 1.5 mm) (P<.001). Significant interactions were present in $L^{\ast}a^{\ast}b^{\ast}$ values between the number of firings and ceramic thickness (P<.001). An increase in number of firings resulted in significant increase in $L^{\ast}$ values for both 0.5 mm and 1.5 mm thicknesses (P<.01, P=.013); however it decreased for 1 mm thickness (P<.01). The $a^{\ast}$ values increased for 1 mm and 1.5 mm thicknesses (P<.01), while it decreased for 0.5 mm specimens. The $b^{\ast}$ values increased significantly for all thicknesses (P<.01, P=.022). As the dentin ceramic thickness increased, significant reductions in $L^{\ast}$ values (P<.01) were recorded. There were significant increases in both $a^{\ast}$ and $b^{\ast}$ values (P<.01) as the dentin ceramic thickness increased. CONCLUSION. The number of firings and dentin ceramic thickness have a definite effect on the final color of all ceramic system tested. The mean ${\Delta}E$ value increased as the dentin ceramic thicknesses increased for zirconium-oxide based all ceramic specimens tested. However, the mean ${\Delta}E$ values were less than 3.7${\Delta}E$ units which is rated as a match in the oral environment.