• Title/Summary/Keyword: In vitro screening

검색결과 586건 처리시간 0.034초

신규 분리균주 Streptomyces sp. BCNU 1030의 in vitro 항균활성 (In vitro Antimicrobial Activity of a New Isolate Streptomyces sp. BCNU 1030)

  • 방지훈;최혜정;안철수;김동완;정영기;주우홍
    • 생명과학회지
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    • 제21권4호
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    • pp.589-595
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    • 2011
  • 질병를 유발하는 미생물들의 항생물질에 대한 증가하는 내성문제를 극복할 수 있는 새로운 항생물질을 개발하고자 항생물질을 생산하는 방선균을 탐색하여 특성화하는데 본 연구는 초점을 맞추고 있다. 충청북도 일대의 토양으로부터 PDA, starch casein agar 등의 배지를 이용하여 100여 개의 방선균주를 순수분리하였다. 순수분리된 방선균주 중 메티실린 내성 황색포도상구균에 가장 활성이 뛰어난 BCNU 1030을 선발하여 형태학적, 생화학적, 분자생물학적 방법으로 동정한 결과 Streptomyces 속으로 동정되었다. 분리균주인 BCNU 1030은 MRSA를 포함한 그람양성 및 그람음성세균에 대한 폭넓은 항균스펙트럼을 가졌다. BCNU 1030의 디클로로메탄 추출물은 MRSA CCARM 3090에 대하여 최소$0.78\;{\mu}g/ml$의 농도에서 저해활성이 있음이 확인되었다. 그러므로 BCNU 1030은 새로운 anti-MRSA 항생물질의 개발에 있어서 가능성을 가지고 있는 것으로 판단된다.

Hormonal Effects of Several Chemicals in Recombinant Yeast, MCF-7 Cells and Uterotrophic Assays in Mice

  • Park, Jin-Sung;Lee, Beom-Jun;Kang, Kyung-Sun;Tai, Joo-Ho;Cho, Jae-Jin;Cho, Myung-Haing;Inoue, Tohru;Lee, Yong-Soon
    • Journal of Microbiology and Biotechnology
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    • 제10권3호
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    • pp.293-299
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    • 2000
  • Many methods have been developed for screening chemicals with hormonal activity. Using recombinant yeasts expressing either human estrogen receptor [Saccharomyces cerevisiae ER + LYS 8127 (YER)] or androgen receptor [S. cerevisiae AR + 8320 (YAR)], we evaluated the hormonal activities of several chemicals by induction of ${\beta}-galactosidase$ activity. The chemicals were $17{\beta}-estradiol$ (E2), testosterone (T), ${\rho}-nonylphenol$ (NP), bisphenol A (BPA), genistein (GEN), 2-bromopropane (2-BP), dibutyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and butylparaben (BP). To assess the estrogenicity of NP, the result of the in vitro recombinant yeast assay was compared with an E-screen assay using MCF-7 human breast cancer cells and an uterotrophid assay using ovariectomized mice. In the YER yeast cells, E2, NP, BPA, GEN, and BP exhibited estrogenicity in a doseresponse manner, while TCDD did not. All the chemicals tested, except T, did not show androgenicity in the YAR yeast cell. The sensitivity of the yeast (YER) assay system to the estrogenic effect of NP was similar to that of the E-screen assay. NP was also estrogenic in the uterotrophic assay. However, in terms of convenience and costs, the yeast assay was superior to the E-screen assay or uterotrophic assay. These results suggest that the recombinant yeast assay can be used as a rapid tool for detecting chemicals with hormonal activities.

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HtrA2 Interacts with Aβ Peptide but Does Not Directly Alter Its Production or Degradation

  • Liu, Meng-Lu;Liu, Ming-Jie;Kim, Jin-Man;Kim, Hyeon-Jin;Kim, Jeong-Hak;Hong, Seong-Tshool
    • Molecules and Cells
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    • 제20권1호
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    • pp.83-89
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    • 2005
  • HtrA2/Omi is a mammalian mitochondrial serine protease homologous to the E. coli HtrA/DegP gene products. Recently, HtrA2/Omi was found to have a dual role in mammalian cells, acting as an apoptosis-inducing protein and being involved in maintenance of mitochondrial homeostasis. By screening a human brain cDNA library with $A{\beta}$ peptide as bait in a yeast two-hybrid system, we identified HtrA2/Omi as a binding partner of $A{\beta}$ peptide. The interaction between $A{\beta}$ peptide and HtrA2/Omi was confirmed by an immunoblot binding assay. The possible involvement of HtrA2/Omi in $A{\beta}$ peptide metabolism was investigated. In vitro peptide cleavage assays showed that HtrA2/Omi did not directly promote the production of $A{\beta}$ peptide at the ${\beta}/{\gamma}$-secretase level, or the degradation of $A{\beta}$ peptide. However, overexpression of HtrA2/Omi in K269 cells decreased the production of $A{\beta}40$ and $A{\beta}42$ by up to 30%. These results rule out the involvement of HtrA2/Omi in the etiology of Alzheimer's disease. However, the fact that overexpression of HtrA2/Omi reduces the generation of $A{\beta}40$ and $A{\beta}42$ suggests that it may play some positive role in mammalian cells.

HeLa 세포에서 IgE-dependent Histamine-Releasing Factor와 Triosephosphate Isomerase의 상호작용 규명 (Interaction between IgE-Dependent Histamine-Releasing Factor and Triosephosphate Isomerase in HeLa Cells)

  • 문지애;김화정;이경림
    • 한국미생물·생명공학회지
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    • 제33권4호
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    • pp.255-259
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    • 2005
  • 본 실험에서는 HRF의 조절단백질을 알아보기 위해 HRF를 bait로 한 yeast two hybrid assay를 실행한 결과 해당과정에 관여하는 TPI(triosephosphate isomerase)라는 효소를 발견하였으며, 가장 많이 중복되어 있었다. In vitro에서 HRF는 TPI의 C말단 잔기 부근(아미노산 156-249)이 상호작용에 주로 관여하는 부위임을 알 수 있었다. 또한, HeLa 세포에서 immunoprecipitation을 이용하여 HRF와 TPI의 상호작용이 실제 in vivo에서도 일어나는 현상이라는 것을 밝혔다. 결과적으로 HRF와 TPI 상호작용은 세포내 일정량이 존재하며 여러가지 신호전달에 의해 동시에 Na,K-ATPase와도 상호작용하는 것으로 생각된다.

Angiotensin II $AT_1$ 수용체 길항제인 SK-1080의 적출심장에 대한 허혈후 재관류시의 작용 및 혈소판응집과 혈액응고에 대한 효과 (Effects of the AngiotensinII $AT_1$ Receptor Antagonist SK-1080 on Ischemia/reperfusion in Isolated Rat Hearts and on Platelet Aggregation and Coagulation in Human Blood)

  • 우수경;최상수;이병호;권광일
    • 약학회지
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    • 제44권6호
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    • pp.558-565
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    • 2000
  • SK-1080 is one of the newly developed orally active nonpeptide angiotensinII $AT_1-receptor$ antagonist that selectively acts at $AT_1$ receptor with high affinity. The cardiac effect on ischemia/reperfusion injury of SK-1080 was compared with those of losartan, a prototype of this class, in isolated rat hearts. Isolated perfused rat heart was pretreated with drug for 10 min and then subjected to global ischemia for 30 min followed by reperfusion with- or without drug for 30 min. The possible additive effect of SK-1080 on the platelet aggregation and coagulation in human blood was also studied. We investigated whether SK-1080 effects the platelet aggregation induced by ADP, a platelet agonist partially dependent on $thromboxaneA_2$. The clotting times in the prothrombin time (PT) and activated partial thromboplastin time (APTT) were also examined in human plasma in vitro as coagulation screening test. SK-1080 improved reperfusion function (LVDP, left ventricular developed pressure; PRP, rate-pressure product) in a dose-dependent manner. SK-1080 reduced ADP-induced platelet aggregation compared with vehicle but less than losartan, and did not affect clotting times.

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Growth-Inhibiting Effects of Vegetable Extracts on Beneficial and Harmful Human Intestinal Bacteria

  • Kim, Moo-Key;Kim, Min-Jeong;Shin, Dong-Hwa;Song, Chul-Gyu;Lee, Hoi-Seon
    • Journal of Applied Biological Chemistry
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    • 제44권2호
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    • pp.65-70
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    • 2001
  • Ethanol extracts of 38 vegetables were subjected to an in vitro screening for their growth-inhibitory activities towards Bifidobacterium bifidum, B. longum, Clostridium perfringens, Lactobacillus acidophilus, L. casei, and Escherichia coli using paper disc agar diffusion methods under anaerobic conditions. The responses varied with both bacterial strain and vegetable species. In a test with 20 mg/disc, Zingiber officinale extracts showed significant growth-inhibitory responses against B. bifidum, and strong inhibitions against L. casei were detected in the extracts of Chrysanthemum coronarium var. spatiosum and Lactuca sativa. The extracts of Allium sativum, Capsicum annuum, L. esculentum, L. esculentum var. cerasiforme, and Z. officinale showed strong inhibitory activities against C. perfringens, while moderate growth-inhibitory responses were observed in the extracts of C. ffutescens, Cucurbita moschata, Daucus carota var. sativa, and Rubus coreanus. However, all vegetable extracts showed no inhibitions against B. longum, L. acidophilus, and E. coli. In tests with 5 mg/disc, moderate inhibitions were observed in the extracts of C. coronarium var. spatiosum and L. sativa against L. casei and Z. officinale against B. bifidum. Vegetables extracts, except for C. coronarium var. spatiosum, L. sativa, and Z. officinale, did not affect the growth of beneficial bacteria. Strong inhibitory responses against C. perfringens were detected in the extracts of C. annuum and L. esculentum var. cerasiforme. Daily intake of vegetables may be important in the prevention of human diseases caused by the intestinal bacteria.

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식이 뽕잎이 흰쥐의 장내균총 조성에 미치는 영향 (Effect of Dietary Mulberry leaf on the Composition of Intestinal Microflora in SD Rats)

  • 이희삼;전호정;이상덕;문재유;김애정;류강선
    • 한국식품과학회지
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    • 제33권2호
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    • pp.252-255
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    • 2001
  • 뽕잎 분말이 1, 10%가 되도록 사료에 첨가시켜 4주동안 SD 흰쥐에게 급여한 후 이들의 대장 내용물로부터 총균수와 주요장내미생물의 생균수를 배지를 이용하여 혐기배양한 후 측정한 결과 총균수는 뽕잎 투여군에서 대조군에 비해 감소하는 경향을 보였다. 그리고 유해균으로 분류할 수 있는 Clostridium은 대조군보다 뽕잎투여군에서 유의성있게 감소하였고, E. coli와 Staphylococcus도 감소하는 경향을 보였다. 뽕잎 메탄올 추출물과 분획물들의 미생물의 생장억제효과를 측정한 결과, 헥산분획과 물분획물이 Clostridium perfringens에 대하여 억제효과를 보였다. 장내균총의 개선 측면에서 뽕잎은 기능성 식품소재로서의 이용가능성이 있는 것으로 나타났다.

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림포사이트의 증식과 대식세포의 활성화에 관한 린나린과 아세칠 린나린의 비교 연구 (Comparison of Linarin and Its Acetylated linarin for the Cytotoxicity on the Proliferation of Lymphocytes and Activation of Macrophages)

  • 한신하;신지혜;소명아;한혜란;김민준;임동술;김경제
    • 약학회지
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    • 제47권4호
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    • pp.234-238
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    • 2003
  • Linarin is a main compound from Chrysanthemum zawadskii var, latilobum. However, the biological mechanisms of these activities are unclear. Because of this wide diversity of effects, it is believed that they may be exerted through pluripotent effectors of linarin. In our previous screening study, the effects of linarin on the mouse macrophages cell line, RAW 264.7 cells, were investigated. It was found that linarin could stimulate macrophages activation by the production of tumor necrosis factor (TNF). The linarin (6.25∼12.5 $\mu\textrm{g}$/mι) inhibited the production of NO in LPS-activated RAW 264.7 cells and linarin became an useful candidates for the development of new drug to treat endotoxemia and inflammation accompanied by the overproduction of NO. However, linarin-treated total lymphocyte showed cytotoxicity in a dose dependent manner between 20 $\mu\textrm{g}$/mι and 40 $\mu\textrm{g}$/mι. In this study, linarin derivative (acetylated linarin) was synthesized in order to obtain less-cytotoxicity of linarin and evaluated for their in vitro cytotoxic activity aganist mouse total lymphocyte. There was no cytotoxic activity in a dose dependent manner (20∼40 $\mu\textrm{g}$/mι) of acetylated linarin whereas linarin showed. The production of NO, however, was not the case by this modified linarin. The cell morphological change was not significantly changed in response to acetylated linarin alone and these effects were potentiated by the addition of LPS. These results suggest that acetylated linarin may be developed to be a promising new drug candidate without cytotoxicity on the basis of its activity of macrophage activation.

시험관내에서 천연물제제 BSASM의 항염증 및 면역억제 효능 평가 (Evaluation of the Anti-inflammatory and Immunomodulatory Effects of BSASM Using in vitro Experiments)

  • 이종성;박유미;박병화;정광선;김국현;이원희;박덕훈
    • 생약학회지
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    • 제34권3호통권134호
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    • pp.228-232
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    • 2003
  • For effective management of atopic dermatitis, it is important to introduce a therapeutic agent although having the fewest side effects, has the greatest anti- inflammatory effect. In the course of screening anti-inflammatory agents, we obtained BSASM composed of several plant extracts. This study was designed to investigate anti-inflammatory and immunomodulatory effects of BSASM. As a first step, $NF-{\kappa}B$ luciferase reporter assay was performed to know the involvement of BSASM in the production of proinflammatory cytokines because $NF-{\kappa}B$ element has been known to play a major role in expression of cytokine genes such as interleukin-8 (IL-8) or tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$. LPS (lipolysaccharide)-induced $NF-{\kappa}B$ activation was inhibited by BSASM. In addition, we found the fact that BSASM inhibits LPS-induced produced production of IL-8 and $TNF-{\alpha}$ proinflammatory cytokines, indicating BSASM has anti-inflammatory effect. In interleukin-2 (IL-2) luciferase reporter assay in Jurkat T cells, BSASM reduced PHA (Phytohemagglutinin)-induced IL-2 luciferase activity, suggesting the possibility that BSASM might also have an immunomodulatory function in T cell-mediated immune response. Based on these results, we suggest the possibility that BSASM can be introduced to improve symptom of immune-related skin diseases, namely, atopic dermatitis.

Comparison of minimally invasive versus conventional open harvesting technique for iliac bone graft in secondary alveolar bone grafting in cleft palate patients: a systematic review

  • Saha, Aditi;Shah, Sonal;Waknis, Pushkar;Bhujbal, Prathamesh;Aher, Sharvika;Vaswani, Vibha
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제45권5호
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    • pp.241-253
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    • 2019
  • This study evaluated and compared the donor site morbidity following minimally invasive and conventional open harvesting of iliac bone for secondary alveolar bone grafting in cleft palate patients. A thorough electronic search of PubMed, Google Scholar, EMBASE, and an institutional library and manual search of various journals was done; Inclusion criteria: 1) full-text articles using a minimally invasive or conventional open harvesting technique for iliac bone for secondary alveolar grafting in cleft palate patients and 2) articles published between January 1, 2001 and June 30, 2017 and Exclusion criteria: 1) articles published in languages other than English, 2) case reports, case series, animal studies, in vitro studies, and letters to the editor, and 3) full-text article unavailable even after writing to the authors. Preliminary screening of 274 studies excluded 223 studies for not meeting the eligibility criteria. Of the remaining 51 studies, 19 were removed for being duplicates. Of the remaining 32 studies, 15 were excluded after reading the abstract. Of the 17 studies that were left, 2 were excluded because they were in a language other than English, and 2 were excluded because the study group did not mention cleft palate patients. Thus, 13 studies providing results for a total of 654 patients were included in this qualitative synthesis. Minimally invasive bone graft harvest techniques are better than the conventional open iliac bone harvest method because they offer shorter operative time, decreased requirement for pain medications, less pain on discharge, and a shorter hospital stay.