• 제목/요약/키워드: cell lysis

검색결과 217건 처리시간 0.022초

Cooperation between Human DAF and CD59 in Protecting Cells from Human Complement-mediated Lysis

  • Xu, Li;Wu, Wenlan;Zhao, Zhouzhou;Shao, Huanjie;Liu, Wanhong;Liu, Hui;Li, Wenxin
    • BMB Reports
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    • 제39권6호
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    • pp.743-748
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    • 2006
  • The complement (C) regulatory proteins decay accelerating factor (DAF, CD55) and CD59 could protect host cells using different mechanisms from C-mediated damage at two distinct levels within the C pathway. Co-expression of DAF and CD59 would be an effective strategy to help overcome host C-induced xenograft hyperacute rejection. In this study, we made a construct of recombinant expression vector containing DAF and CD59 cDNA and the stable cell lines were obtained by G418 selection. Extraneous genes integration and co-expression were identified by PCR, RT-PCR and Western blot analysis. Human c-mediated cytolysis assays showed that NIH/3T3 cells transfected stably with pcDNA3-CD59, pcDNA3-DAF, and pcDNA3-CD59DAF-DP were protected from C-mediated damage and that synchronously expressed human CD59 and DAF provided the most excellent protection for host cells as compared with either human CD59 or DAF expressed alone. Therefore, the construct represents an effective and efficacy strategy to overcome C-mediated damage in cells and, ultimately, in animals.

Cross-immunizing potential of tumor MAGE-A epitopes recognized by HLA-A*02:01-restricted cytotoxic T lymphocytes

  • Huang, Ze-Min;Jia, Zheng-Cai;Tang, Jun;Zhang, Yi;Tian, Yi;Ni, Dong-Jing;Wang, Fang;Wu, Yu-Zhang;Ni, Bing
    • BMB Reports
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    • 제45권7호
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    • pp.408-413
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    • 2012
  • Almost all melanoma cells express at least one member of the MAGE-A antigen family, making the cytotoxic T cells (CTLs) epitopes with cross-immunizing potential in this family attractive candidates for the broad spectrum of anti-melanoma immunotherapy. In this study, four highly homologous peptides (P264: FLWGPRALA, P264I9: FLWGPRALI, P264V9: FLWGPRALV, and P264H8: FLWGPRAHA) from the MAGE-A antigens were selected by homologous alignment. All four peptides showed high binding affinity and stability to HLA-A$^*02:01$ molecules, and could prime CTL immune responses in human PBMCs and in HLA-A$^*02:01/K^b$ transgenic mice. CTLs elicited by the four epitope peptides could cross-lyse tumor cells expressing the mutual target antigens, except MAGE-A11 which was not tested. However, CTLs induced by P264V9 and P264I9 showed the strongest target cell lysis capabilities, suggesting both peptides may represent the common CTL epitopes shared by the eight MAGE-A antigens, which could induce more potent and broad-spectrum antitumor responses in immunotherapy.

Isolation, Physiological Characterization of Bacteriophages from Enhanced Biological Phosphorus Removal Activated Sludge and Their Putative Role

  • Lee, Sang-Hyon;Satoh, Hiroyasu;Katayama, Hiroyuki;Mino, Takashi
    • Journal of Microbiology and Biotechnology
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    • 제14권4호
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    • pp.730-736
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    • 2004
  • This study aims at characterizing the bacteriophages isolated from activated sludge performing enhanced biological phosphorous removal (EBPR) to understand the interactions between the phage-host system and bacterial community. Sixteen bacterial isolates (E1-E16) were isolated as host bacterial strains from EBPR activated sludge for phage isolation. Forty bacteriophages based on their plaque sizes (2 plaques on E4, 4 on E8, 11 on E10, 5 on E14, 18 on E16) were obtained from filtered supernatant of the EBPR activated sludge. Each bacteriophage did not make any plaque on bacterial strains tested in this study except on its own host bacterial strain, respectively, indicating that the bacteriophages are with narrow host specificity. However, fourteen of the forty bacteriophages obtained in this study lost their virulent ability even on their own host bacteria. All of the lytic phages showed similar one-step growth patterns and had long latent period (about 9 hours) to reproduce their phage particles in their host bacterial cells. On the other hand, their probable burst sizes (6 to 48 per host cell) were large enough to actively lyse their host bacterial cells. Therefore, it could be implied that bacteriophages are also important members of the microbial community in EBPR activated sludge, and lytic phages directly decrease the population size of their host bacterial groups in EBPR activated sludge by lysis.

관형막을 이용한 혐기소화조의 전처리 농축공정으로써 적용가능성 연구 (Application of tubular membrane to thickening process as a pre-treatment of anaerobic digester)

  • 강희석;박기태;박정우;김형수
    • 상하수도학회지
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    • 제29권2호
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    • pp.203-209
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    • 2015
  • The purpose of this study is to estimate the applicability of a stable anaerobic digester using a separator membrane to the preprocessing thickening process. The results of the experiments showed about a 47.16% weight loss rate for the sludge under anaerobic condition, and about 41.17% under intermittent aeration condition. The concentrations of rejection water were $SCOD_{Cr}$ 25 mg/L, T-N 16.6 mg/L, and T-P 1.4 mg/L on the average under the intermittent aeration condition, which were lower than the concentrations of rejection water under an anaerobic condition. As for the factors affecting the reduction of the flux, correlation analyses of TTF, MLSS, $SCOD_{Cr}$, and $EPS_{Protein}$, $EPS_{Polysacchride}$ resulted in -0.97, -0.95, -0.84 and -0.86, -0.95, respectively, which showed that TTF had the highest correlation to the reduction of the flux. In addition, it was concluded that MLSS, $SCOD_{Cr}$ and $EPS_{Protein}$, $EPS_{Polysacchride}$ also have close correlations. The results are considered to show that, in the case of the process using a tubular membrane in the preprocessing process of an anaerobic digester, an intermittent aeration condition of the thickener considering the contamination of the membrane and load of rejection water is appropriate for the stable preprocessing process.

Streptomyces subrutilus P5가 생산하는 철 함유 superoxide dismutase의 분비 (Secretion of the iron containing superoxide dismutase of Streptomyces subrutilus P5)

  • 박재승;김재헌
    • 미생물학회지
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    • 제51권2호
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    • pp.108-114
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    • 2015
  • 본 연구에서는 Streptomyces subrutilus P5의 생장과 세포내 외 철 함유 superoxide dismutase 활성을 비교 분석하여 철함유 superoxide dismutase의 분비 시점을 확인하고 분자 수준에서 이 효소의 분비에 관여하는 유전정보를 확인하고자 하였다. Streptomyces subrutilus P5의 균체 생장은 건체 중량을 측정하여 결정하였다. Glucose는 log phase에서 급격히 소모되어 24시간 후에 이르러 완전히 고갈되었다. 세포내의 철 함유 superoxide dismutase는 배양 후 3시간에 나타나며 세포외 철 함유 superoxide dismutase는 배양 후 7.5시간부터 나타난다. 따라서 superoxide dismutase는 용균에 의해서가 아니라 능동적인 분비기작에 의해서 세포 외로 분비된 것으로 추측할 수 있다. Streptomyces subrutilus P5의 sodF에는 signal peptide 유전정보가 존재하지 않았다. 그러나 sodF의 상류지역에서 다른 세균의 type III 분비단백질 유전자와 유사한 type III 분비상자가 발견되었다. Streptomyces 균주에서 type III 분비단백질이 존재할 가능성이 있음을 처음으로 제시하였다.

이온교환크로마토그래피에서 라이소자임 분리에 미치는 pH와 온도 영향 (The Effect of pH and Temperature on Lysozyme Separation in Ion-exchange Chromatography)

  • 고관영;김인호
    • Korean Chemical Engineering Research
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    • 제52권1호
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    • pp.98-105
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    • 2014
  • 라이소자임은 용균 작용, 조직 회복 과정의 촉진 등의 작용을 하며 난백 중에 0.3% 함유되어 있다. 난백에서 라이소자임을 분리하는 방법으로 친화성 크로마토그래피, 이온교환크로마토그래피, 한외여과법 등이 있는데 이 중 이온교환크로마토그래피가 가장 많이 사용된다. 라이소자임을 양이온 젤이 충전된 유리 칼럼에서 분리 정제할 때 최적의 pH와 온도 조건을 찾는 것을 실험 목표로 하고, ASPEN Chromatography 전산 모사의 결과와 비교하였다. 실험에 사용되는 완충용액은 인산완충용액이었고 pH를 5~8로 변화를 주어 상온에서 실험하였고, 가장 분리가 잘 이루어진 pH에서 온도를 $5^{\circ}C$ 간격으로 $25{\sim}40^{\circ}C$로 변화시켜 실험하였다. RP-HPLC (Reversed phase High Performance Liquid Chromatography) 분석을 통해 라이소자임의 체류 시간을 확인하였고, OriginPro 8을 이용해 용출 단계에서 크로마토그램의 면적을 비교하여 라이소자임의 양을 정량분석하였다. 결과를 분석한 결과, pH 5일 때, 온도가 $25^{\circ}C$에서 가장 많은 양의 라이소자임이 분리되었다.

인삼뿌리 썩음 병균 Fusarium solane에 대한 Streptomyces species의 길한작용 (Antagonistic activity of Streptomyces apecies against Fusarium solani causing ginseng root rot)

  • 정영륜;오승환;정후섭
    • 미생물학회지
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    • 제27권1호
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    • pp.56-62
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    • 1989
  • Chitin 분해 활성이 높은 Streptomyces 균주들을 인삼재배 토양에서 분리하고, 그 균주들이 인상뿌리 썩음 병균인 Fµsarium solani 에 미치는 길항효과와 그 작용을 조사하였다. 분리된 Streptomyces 중 몇 균주는 F. solani 의 포자발아와 발 아판의 생장을 억제하였고 균사를 분해하였으며, 동시에 세포벽 분해효소로 알려친 Chitinase를 생산하였다. 그중에서도 S alboniger ST 59 와 S. roseolilacinus ST 129 는 길항효과가 아주 좋았는데, 병원균의 분생포자를 두 Streptomyces 의 농축배 양여액에 14일간 처리하였을 때 포지숫자가 처음 정종 정도의 20%로 줄어을였다. 특히 S. alboniger ST 59는 뱅균인 F solani의 분생포자, 균사 뿐만 아니라 분해되기 어려운 후막포자까지도 분해하였다. 이것으로 비푸어 보건데, F. solani의 이 Streptomycetes에 의한 억제작용은 항생물질에 영향을 받은 병균이 Chitinase에 의해 분해된 것으로 생각된다.

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Cecropin A-Magainin 2 유도체 펩티드의 Trichosporon beigelii에 대한 항진균 활성 및 인간 적혈구 세포에 대한 용혈활성 (Fungicidal and Hemolytic Activity of Cecropin A-Magainin 2 Analogue Peptides against Tri-chospoon beigelii and Human Red Blood Cells)

  • 이동건;신송엽;이명규;함경수
    • 미생물학회지
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    • 제33권3호
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    • pp.170-174
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    • 1997
  • 강한 항진균 활성을 나타내며, 낮은 적혈구 용혈활성을 갖는 새로운 합성 펩티드를 얻기 위하여 cecropin A(1-8)-magainin 2(1-12) 및 이들의 유사체들을 설계하고 이들을 고상법에 의하여 합성하였다. 합성 펩티드의 항진균 활성은 Trichosporon beigelii에 대한 성장억제능에 의하여 측정하였으며, 세포독성은 인간의 적혈구 세포에 대한 용혈활성에 의하여 측정하였다. 펩티드의 양쪽 친매성의 증가는 항진균활성보다는 적혈구 용혈황성에 큰 영향을 미쳤다. Cecropin A(1-8)-magainin 2(1-12)의 12번 아미노산이 Lys을 Ala으로 치환시킨 유사체 펩티드(A2)는 가장 강한 항진균활성(minimum inhibitory concentration : 2.5.$\mu$g/ml)을 나타내며 비교적 낮은 적혈구 용혈활성(200.$\mu$g/ml의 펩티드 농도에서 0.5% hemolysis를 나타냄)을 나타내었다. 따라서 A2의 펩티드는 세포독성을 갖지 않으며 강한 항진균 활성을 가지는 모델로서 유용하게 사용될 것이다.

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적조생물 Cochlodinium polykrikodes 구제효율에 미치는 황토의 광물학적 특징 (Effects of Yellow Clay Contents on Removal Efficiency of Harmful Dinoflagellate Cochlodinium polykrikoides)

  • 박영태;박호섭;박태규;안경호;손문호;김평중;박맹언
    • 수산해양교육연구
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    • 제28권6호
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    • pp.1662-1672
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    • 2016
  • To address physicochemical factors of yellow clay for removing Cochlodinium polykrikoides blooms, the correlation of removal efficiency with mineralogical characteristics of yellow clay collected from various areas was surveyed. Yellow clay from different localities showed the wide range of chemical compositions $SiO_2$ : 43~71%, $Al_2O_3$ : 13~26%, $Fe_2O_3$ : 5~14%, MgO : 0.4~1.8%, $K_2O$ : 0.6~3.3%, L.O.I.(Loss of Ignition) : 4.5~15%. The mineral compositions of yellow clay were mainly consisted of quartz and feldspar including small amounts of kaolinite, chlorite, and Fe-oxides. The result of size analysis showed that $6{\Phi}(31{\sim}16{\mu}m)$ and $7{\Phi}(16{\sim}8{\mu}m)$ were dominated sizes. The zeta-potential were in the range of -4.1~-20.7mV(average -13.7). As increasing removal efficiency of C. polykrikoides, contents of $Al_2O_3$, $Fe_2O_3$ and L.O.I. in yellow clay increased, whereas $SiO_2$ content decreased. Furthermore, the amounts of silt mineral and small particle were high when the removal efficiency was high. According to factor analysis using principle component analysis, two components of factor 1 and factor 2 showed 79% of the total variance, which is related to cohesion and adsorption. Inducing cell lysis of C. polykrikoides by cohension and adsorption between C. polykrikoides and yellow clay.

홍삼산성다당체 (RGAP)의 경구투여에 의한 항종양 효과 (Effect of Oral Administration of Red ginseng acidic polysaccharide (RGAP) on the Tumor Growth Inhibition)

  • 곽이성;신한재;송용범;경종수;위재준;박종대
    • Journal of Ginseng Research
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    • 제29권4호
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    • pp.176-181
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    • 2005
  • Our previous reports demonstrated that ip. administration of Korean red ginseng acidic polysaccharide (RGAP) exerts antitumor activity In mice. The present study was carried out to compare the effects of ip. and p.o. routes of administration of RGAP on either normal or tumor-bearing BALB/c mice. RGAP was administered either ip. or p.o. at doses of 100, 300, 500, 1000 mg/kg for 1 or 5 weeks. Peritoneal macrophages from mice treated with RGAP p.o. at a dose of 300 mg/kg either for 1 or 5 weeks did not exhibit growth inhibition activity toward WEHI-I64 tumor cells. However, administration of RGAP at a dose of 600 mg/kg for both 1 and 5 weeks increased the antitumor activity of macrophages. Oral administration of RGAP (600 mg/kg) for 5 weeks and ip. administration of RGAP (300 mg/kg) for 1 week resulted in antitumor activities of $40\%$ and $45\%$, respectively, indicating that the effect of i.p. injection is more potent 2 and 5 times than that of p.o. one in terms of dose and duration, respectively. Tumor inhibition rates of RGAP at doses of 300, 500, 1000 mg/kg in mice transplanted with B16-F10 melanoma were 4.4, 12.0, and $45.4\%$, respectively, meaning that p.o. dose higher than 500 mg/kg possess marked antitumor activity. The results above suggests that p.o. administration of RGAP also show antitumor activity in vivo depending on the dose.