• 제목/요약/키워드: 3D co-culture

검색결과 233건 처리시간 0.027초

천적유지식물로써 괭이밥에 대한 기초연구: 괭이밥의 귤응애 밀도억제효과와 천적 2종의 괭이밥응애 포식력 (A preliminary study of Oxalis corniculata L. as a new banker plant: Control efficacy against Panonychus citri (McGregor) and feeding ability of two natural enemies on Tetranychina harti (Ewing))

  • 오창학;진혜영;안태현;송유진;전혜정;이준석;함은혜
    • 한국응용곤충학회지
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    • 제56권3호
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    • pp.267-273
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    • 2017
  • 귤나무속 과수의 주요 해충인 귤응애의 방제효과 극대화를 위해 괭이밥의 천적유지식물 적용 가능성을 검토하였다. 괭이밥을 지피식물로 조성한 처리구에서 일반 초생재배구에 비해 포식성 응애류 밀도가 평균 3배 이상 높게 나타났으며, 제초구에 비해 3.7배 낮은 귤응애 발생 밀도를 확인 할 수 있었다. 칠레이리응애는 괭이밥응애 알과 성충을 포식하지 못하였고, 사막이리응애는 일일 평균 괭이밥응애 알 1.96개와 암컷 성충 1.93개를 포식하였다. 온도 $23{\pm}1^{\circ}C$, 습도 $70{\pm}5%$, 광조건 16L : 8D로 설정된 사육실에서 괭이밥응애 알에서 성충까지 17.4일이 소요되었으며 77.5%의 부화율과 84.8%의 유충생존율을 확인하였다.

Extracellular 5-Aminolevulinic Acid Production by Escherichia coli Containing the Rhodopseudomonas palustris KUGB306 hemA Gene

  • Choi, Han-Pil;Lee, Young-Mi;Yun, Cheol-Won;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • 제18권6호
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    • pp.1136-1140
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    • 2008
  • The Rhodopseudomonas palustris KUGB306 hemA gene codes for 5-aminolevulinic acid (ALA) synthase. This enzyme catalyzes the condensation of glycine and succinyl-CoA to yield ALA in the presence of the cofactor pyridoxal 5'-phosphate. The R. palustris KUGB306 hemA gene in the pGEX-KG vector system was transformed into Escherichia coli BL21. The effects of physiological factors on the extracellular production of ALA by the recombinant E. coli were studied. Terrific Broth (TB) medium resulted in significantly higher cell growth and ALA production than did Luria-Bertani (LB) medium. ALA production was significantly enhanced by the addition of succinate together with glycine in the medium. Maximal ALA production (2.5 g/l) was observed upon the addition of D-glucose as an ALA dehydratase inhibitor in the late-log culture phase. Based on the results obtained from the shake-flask cultures, fermentation was carried out using the recombinant E. coli in TB medium, with the initial addition of 90 mM glycine and 120 mM succinate, and the addition of 45 mM D-glucose in the late-log phase. The extracellular production of ALA was also influenced by the pH of the culture broth. We maintained a pH of 6.5 in the fermenter throughout the culture process, achieving the maximal levels of extracellular ALA production (5.15 g/l, 39.3 mM).

Statistical optimization of phytol and polyunsaturated fatty acid production in the Antarctic microalga Micractinium variabile KSF0031

  • Kim, Eun Jae;Chae, Hyunsik;Koo, Man Hyung;Yu, Jihyeon;Kim, Hyunjoong;Cho, Sung Mi;Hong, Kwang Won;Lee, Joo Young;Youn, Ui Joung;Kim, Sanghee;Choi, Han-Gu;Han, Se Jong
    • ALGAE
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    • 제37권2호
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    • pp.175-183
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    • 2022
  • Polar microorganisms produce physiologically active substances to adapt to harsh environments, and these substances can be used as biomedical compounds. The green microalga Micractinium variabile KSF0031, which was isolated from Antarctica, produced phytol, a natural antimicrobial agent. Furthermore, several polyunsaturated fatty acids (PUFAs), including omega-3, exhibit antioxidant properties. Here statistical methods (Plackett-Burman design and Box-Behnken design) were used to optimize the culture medium of KSF0031 to improve biomass production, and K2HPO4, MgSO4·7H 2O, and ammonium ferric citrate green (AFCg) were selected as significant components of the culture medium. Changes in the concentration of K2HPO4 and MgSO4·7H 2O as positive factors and AFCg as a negative factor affected cell growth to a remarkable degree. The biomass production in a 100 L culture using the optimized medium for 24 d at 18℃ was improved by 37.5% compared to that obtained using the original BG-11 medium. The quantities of PUFAs and phytol obtained were 13 mg g-1 dry cell weight (DCW) and 10.98 mg g-1 DCW, which represent improved yields of 11.70% and 48.78%, respectively. The results of this study could contribute to an improved production of phytol and fatty acids from Antarctic microalgae in the biomedical industry.

면역크로마토그래피를 이용한 E. coli O157: H7 신속검출 키트의 유효성 평가 (Evaluation of the Efficiency of E. coli O157: H7 Rapid Detection Kit using Immunochromatography)

  • 곽효선;이동하;문희숙;박종석;우건조;김창민
    • 한국식품위생안전성학회지
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    • 제18권3호
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    • pp.118-124
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    • 2003
  • 식품으로부터 다양한 병원 미생물을 신속 검출하기 위하여 다양한 검출 원리를 이요한 키트들이 개발 시판되고 있다. 검사키트는 신속, 정확하고 간단하게 사용할 수 있으므로 검사기관이나 실험실 뿐 아니라 식품회사에서 QC 또는 QA를 수행하기 위하여 사용이 증가되고 있는 추세이다. 이에 본 연구에서는 E. coli 0157:H7의 단클론항체를 이용하여 면역크로마토글래피법에 의해 개발된 E. coli 0157:H7 검출 키트(Donga Co, Korea, D-kit)에 대한 검출감도 및 특이성을 확인하고 식품 시료에 적용 가능성을 평가하였다. 면역크로마토그래피법에 의하여 개발 시판되고 있는 Reveal E. coli 0157:H7 kit (Neogen Co., USA. R-kit)와 VIP EHEC kit(Biocontro Inc., USA. V-kit)를 비교 키트로 사용하였다. E. coli 0157:H7 표준균주를 사용하여 실시한 검출감도 확인시험 결과 R-kit 및 D-kit는 104/m/의 농도에서 양성으로 확인되었고 $10^3$/ml에서도 약한 양성 반응을 보였으나, V-lit는 $10^5$/ml농도로 검출감도가 낮았다. 또한, 배양액을 가열하여 kit에 적용하는 것이 가열하지 않은 경우보다 검출감도를 높일 수 있었다. E. coli 0157:H7 분리 22주, verotoxin 생성 E. coli 7주 E. coli 분리주 40주 중 3주를 제외한 모든 균에서 음성의 결과를 보여 특이성을 확인하였다. 세 키트에 위양성 반응을 보인 것은 E. coli 0157:H19, E. coli 0148:H18 및 Salmonella gallinarium으로 이들 혈청형과 0157:H7 사이에는 유사한 혈청학적 특성이 존재하는 것으로 추정되었다. 이상의 실험결과로 D-kit는 E. coli 0157:H7을 검출하는데 감도 및 특이성 면에서 기존 키트인 R-kit 및 V-kit와 같이 이용한 것으로 확인되었다.

Bio-hybrid dental implants prepared using stem cells with β-TCP-coated titanium and zirconia

  • Safi, Ihab Nabeel;Hussein, Basima Mohammed Ali;Al-Shammari, Ahmed Majeed
    • Journal of Periodontal and Implant Science
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    • 제52권3호
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    • pp.242-257
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    • 2022
  • Purpose: This study investigated periodontal ligament (PDL) restoration in osseointegrated implants using stem cells. Methods: Commercial pure titanium and zirconium oxide (zirconia) were coated with beta-tricalcium phosphate (β-TCP) using a long-pulse Nd:YAG laser (1,064 nm). Isolated bone marrow mesenchymal cells (BMMSCs) from rabbit tibia and femur, isolated PDL stem cells (PDLSCs) from the lower right incisor, and co-cultured BMMSCs and PDLSCs were tested for periostin markers using an immunofluorescent assay. Implants with 3D-engineered tissue were implanted into the lower right central incisors after extraction from rabbits. Forty implants (Ti or zirconia) were subdivided according to the duration of implantation (healing period: 45 or 90 days). Each subgroup (20 implants) was subdivided into 4 groups (without cells, PDLSC sheets, BMMSC sheets, and co-culture cell sheets). All groups underwent histological testing involving haematoxylin and eosin staining and immunohistochemistry, stereoscopic analysis to measure the PDL width, and field emission scanning electron microscopy (FESEM). The natural lower central incisors were used as controls. Results: The BMMSCs co-cultured with PDLSCs generated a well-formed PDL tissue that exhibited positive periostin expression. Histological analysis showed that the implantation of coated (Ti and zirconia) dental implants without a cell sheet resulted in a well-osseointegrated implant at both healing intervals, which was confirmed with FESEM analysis and negative periostin expression. The mesenchymal tissue structured from PDLSCs only or co-cultured (BMMSCs and PDLSCs) could form a natural periodontal tissue with no significant difference between Ti and zirconia implants, consequently forming a biohybrid dental implant. Green fluorescence for periostin was clearly detected around the biohybrid implants after 45 and 90 days. FESEM showed the invasion of PDL-like fibres perpendicular to the cementum of the bio-hybrid implants. Conclusions: β-TCP-coated (Ti and zirconia) implants generated periodontal tissue and formed biohybrid implants when mesenchymal-tissue-layered cell sheets were isolated from PDLSCs alone or co-cultured BMMSCs and PDLSCs.

The Inhibition of Epileptogenesis During Status Epilepticus by Ginsenosides of Korean Red Ginseng and Ginseng Cell Culture (Dan25)

  • N.E., Chepurnova;Park, Jin-Kyu;O.M., Redkozubova;A.A., Pravdukhina;K.R., Abbasova;E.V., Buzinova;A.A., Mirina;D.A., Chepurnova;A.A., Dubina;U.A., Pirogov;M., De Curtis;L., Uva;S.A., Chepurnov
    • Journal of Ginseng Research
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    • 제31권3호
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    • pp.159-174
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    • 2007
  • Pharmacology of Korean Red ginseng gives us unique possibility to develop new class of antiepileptic drugs today and to improve one's biological activity. The chemical structures of ginsenosides (GS) have some principal differences from well-known antiepileptic new generation drugs. The antiepileptic effect of GS was also demonstrated in all models of epilepsy in rats (young and adult), which have studied, in all models of epilepsy including status epilepticus (SE), induced by lithium - pilocarpine. In our experiments in rats new evidences on protective effects were exerted as a result of premedication by GS. Pre-treatment of several GS could induce decrease of the seizures severity and brain structural damage (by MRI), neuronal degeneration in hippocampus. Wave nature of severity of motor seizures during convulsive SE was observed during lithium-pilocarpine model of SE in rats (the first increase of seizures was 30 min after the beginning of SE and the second - 90 min after. The efficacy of treatment on SE by ginsenoside as expected was observed after no less 3 weeks by daily GS i.p. administration. It is blocked SE or significantly decrease the severity of seizures during SE. The implication of presented data is that combination of ginsenosides from Korean Red ginseng and ginseng cell culture Dan25 that could be applied for prevention of epileptical status development. However, a development of optimal ratio of different ginsenosides $(Rb_1$ Rc, Rg, Rf,) should consummate in the new antiepileptic drug development.

Expression of the Antioxidant Enzyme and Apoptosis Genes in In vitro Maturation/In vitro Fertilization Porcine Embryos

  • Jang, H.Y.;Kong, H.S.;Lee, S.S.;Choi, K.D.;Jeon, G.J.;Yang, B.K.;Lee, C.K.;Lee, H.K
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권1호
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    • pp.33-38
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    • 2004
  • This study was aimed at testing the gene expression of antioxidant enzymes and apoptosis genes for in vitro culture in porcine embryos produced by in vitro maturation/in vitro fertilization (IVM/IVF). Pocine preimplantation embryos obtainted from IVM/IVF can be successfully culture in vitro, but they are delayed or stop to develop at specific developmental stage. Many factors such as reactive oxygen species and apoptosis in an IVM/IVF system followed by in vitro culture influence the rate of production of viable blastocysts. Porcine embryos derived from IVM/IVF were cultured in the atmosphere of 5% $CO_2$ and 20% $O_2$ at $38.5^{\circ}C$ in NCSU23 medium. The patterns of gene expression for antioxidant enzymes and apoptosis genes during in vitro culture in pocine IVM/IVF embryos were examined by the modified semi-quantitative single cell reverse transcriptase-polymerase chain reaction (RT-PCR). Porcine embryos produced by in vitro procedures were expressed mRNAs for CuZn-SOD, GAPDH and GPX, whereas transcripts for Mn-SOD and catalase were not detected at any developmental stages. Expression of caspase-3 mRNA was detected at 2 cell, 8 cell 16 cell and blastocyst, but p53 mRNA was not detected at any stages. The fas transcripts was only detected in blastocyst stage. These results suggest that various antioxidant enzymes and apoptosis genes play crucial roles in vitro culture of porcine IVM/IVF embryos.

GERI-BP001 Compounds, New Inhibitors of Acyl-CoA: Cholesterol Acyltransferase from Aspergillus fumigatus F37

  • Jeong, Tae-Sook;Kim, Sung-Uk;Son, Kwang-Hee;Kwon, Byoung-Mog;Kim, Young-Kook;Bok, Song-Hae
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1995년도 춘계학술대회
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    • pp.67-67
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    • 1995
  • Acyl-CoA:cholesterol acyltransferase (ACAT, EC 2.3.1.26) plays an important role in the control of intracellular free cholesterol content via its cholesterol esterifying activity. ACAT inhibitors are expected to be effective for treatment of atherosclerosis and hypercholesterolemia. In the course of a screening program for ACAT inhibitors from microbial sources, GERI-BP001 M, A, and B were isolated from the fermentation broth of a fungal strain. GERI-BP001 compounds were isolated from a culture broth of Aspergillus fumigatus F37 by acetone extraction, EtOAc extraction, SiO$_2$ column chromatography, and reverse phase HPLC. The structure of GERI-BP001 coumpounds were determined by $^1$H-NMR, $\^$l3/C-NMR, 2D-NMR, NOESY, and long range C-H COSY experiments. GERI-BP001 M, A, and B inhibit ACAT activity in an enzyme assay system using rat liver microsomes by 50% at concentrations of 75, 147, and 71 ${\mu}$M, respectively.

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에키네시아 조직 배양체 추출물의 생리활성에 관한 연구 (Study on Biological Activities of Adventitious Roots Cultured from Echinacea angustifolia)

  • 박창민;정민석;고두진;백기엽;최종완
    • 대한화장품학회지
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    • 제37권3호
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    • pp.219-226
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    • 2011
  • 일반적으로 쌍떡잎식물 국화과 허브의 한 종인 에키네시아는 미국 북미 대평원에 서식하는 야생 식물로 수세기 동안 감기나 또 다른 바이러스 감염에 의한 질병에 대하여 면역 기능을 증진시키는 전통 식물 약제로 널리 사용되어왔다. 본 연구에서는 생물반응장치를 이용하여 조직 배양된 에키네시아 식물체에 대하여 화장품 성분으로써 응용 가치를 평가하였다. 이미 몇몇 보고된 논문에서 에키네시아는 조직재생, 상처 치유, 그리고 염증 억제 등의 약리학적 활성에 대한 연구가 진행되었다. 본 연구에서는 생물반응장치를 이용하여 조직 배양된 에키네시아 추출물에 대하여 화장품 성분으로서 효능, 효과를 평가하였다. 실험 결과 조직배양된 에키네시아 추출물은 항산화 효과 및 단백분해효소 발현 억제 효과가 자연 상태의 에키네시아 추출물과 비교하여 우수한 결과를 보였다. 이러한 결과들은 피부보호를 위한 화장품 성분으로서 응용 가능성을 제공할 수 있을 것으로 사료된다.

Purification and reaction pattern of cephalexin synthesizing enzyme from Acetobacter turbidans

  • Kang, Sang-Moo;Kim, June-Hoe;Kim, Deog-Jung;Kim, Young-Jun
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 1986년도 추계학술대회
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    • pp.525.2-525
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    • 1986
  • Cephalexin synthesizing enzyme (${\alpha}$ amino acid ester hydrolase) was partially purified from the culture broth of Acetobacter turbidans ATCC9325 through ammonium sulfate fractionation, DEAE, CM, and Sephacryl S-200 gel filtration. The enzyme has optimum pH 6.0 and temperature, 40$^{\circ}C$ respectively. From the analysis of reaction mixtures by thin layer chromatographic and high performance liquid chromatographic techniques, it was confirmed this enzyme catalyzed simultaneously the following reactions : 1) Synthesis of cephalexin from D-${\alpha}$-phenylglycine methylester (PGM) and 7-amino 3-deacetoxy-cetoxycephalosporanic acid (7-ADCA) 2) Hydrolysis of cephalexin to form 7-ADCA and phenylglycine (PG) 3) Hydrolysis of PGM to form PG and methanol. Base on the above experimental observations, the reaction model of this enzyme was identical with that of the enzyme from Xanthomonas citri.

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