• 제목/요약/키워드: Adhesion assay

검색결과 226건 처리시간 0.03초

활성화된 단핵구 및 대식세포의 항원제시기능에 대한 Kaempferitrin의 조절 효과 (Modulatory Effect of Kaempferitrin, a 3,7-Diglycosylflavone, on the LPS-Mediated Up-regulation of Surface Co-stimulatory Molecules and CD29-Mediated Cell-cell Adhesion in Monocytic- and Macrophage-like Cells)

  • 김병훈;조동하;조재열
    • 약학회지
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    • 제51권6호
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    • pp.482-489
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    • 2007
  • Kaempferitrin, isolated from Kenaf (Hibiscus cannabinus), was examined to evaluate its modulatory effects on antigen-presenting cell functions of macrophages/monocytes such as phagocytosis of foreign materials, up-regulation of costimulatory molecules (CD40, CD80 and CD86), adhesion molecule activation, and antigen processing and presentation. Kaempferitrin strongly blocked up-regulation of CD40, CD80 and CD86, but not pattern recognition receptor (PRR) (e.g., TLR2). It also suppressed functional activation of CD29 (${\beta}1$-integrins), as assessed by cell-cell adhesion assay, required for T cell-antigen-presenting cell (APC) interaction. Furthermore, this compound did not block a simple activation of CD29, as assessed by cell-fibronectin adhesion assay. However, the compound did not diminish phagocytic uptake, an initial step for antigen processing, and ROS generation in RAW264.7 cells. In particular, to understand molecular mechanism of kaempferitrin-mediated inhibition, the regulatory role of LPS-induced signaling events was examined using immunoblotting analysis. Interestingly, this compound dose dependently suppressed the phosphorylation of $I{\kappa}B{\alpha}$, Src, Akt and Syk, demonstrating that it can negatively modulate the activation of these signaling enzymes. Therefore, our data suggested that kaempferitrin may be involved in regulating APC function-relevant immune responses of macrophages and monocytes by regulating intracellular signaling.

Suppression of Human Fibrosarcoma Cell Metastasis by Phyllanthus emblica Extract in Vitro

  • Yahayo, Waraporn;Supabphol, Athikom;Supabphol, Roongtawan
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권11호
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    • pp.6863-6867
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    • 2013
  • Phyllanthus emblica (PE) is known to exhibit various pharmacological properties. This study aimed to evaluate the antimetastatic potential of a PE aqueous extract. Cytotoxicity to human fibrosarcoma cells, HT1080, was determined by viability assay using the 3-(4,5-dimethylthiazol,2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent. Cell migration and invasion were investigated using chemotaxis chambers containing membranes precoated with collagen IV and Matrigel, respectively. Cell attachment onto normal surfaces of cell culture plates was tested to determine the cell-adhesion capability. The molecular mechanism of antimetastatic activity was assessed by measuring the gene expression of matrix metalloproteinases, MMP2, and MMP9, using reverse transcription-polymerase chain reaction (RT-PCR) assay. The mRNA levels of both genes were significantly down-regulated after pretreatment with PE extract for 5 days. Our findings show the antimetastatic function of PE extract in reducing cell proliferation, migration, invasion, and adhesion in both dose- and time-dependent manners, especially growth arrest with low $IC_{50}$ value. A decrease in the expression of both MMP2 and MMP9 seems to be the cellular mechanism for antimetastasis in this case. There is a high potential to use PE extracts clinically as an optional adjuvant therapeutic drug for therapeutic intervention strategies in cancer therapy or chemoprevention.

In Vitro: Antimicrobial Effect of Lactobacillus salivarius on Staphylococcus pseudintermedius

  • Seo, Haeyoung;Bae, Seulgi;Oh, Taeho
    • 한국임상수의학회지
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    • 제36권2호
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    • pp.98-101
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    • 2019
  • Lactobacillus spp. are the bacteria most commonly used as probiotics and it has been proven that they inhibit pathogenic bacterial growth and improve skin repair in humans. This study was conducted to investigate the growth inhibitory effect of Lactobacillus on Staphylococcus pseudintermedius, the most commonly isolated pathogen in canine pyoderma, and whether Lactobacillus could inhibit the adhesion capability of S. pseudintermedius to canine corneocytes. For this study, L. salivarius and S. pseudintermedius were isolated from healthy beagle fecal samples and the skin surface of dogs with skin infection, respectively. S. pseudintermedius was co-cultured with L. salivarius to assess the inhibitory effect. For the adhesion assay, corneocytes were collected from healthy beagle ventral abdominal skin. Both bacterial species attached to corneocytes and were assessed in number. As a result, L. salivarius significantly inhibited the growth of S. pseudintermedius in the culture medium. Moreover, L. salivarius reduced attachment of S. pseudintermedius in the adhesion assay. These results suggest that L. salivarius has an inhibitory effect on S. pseudintermedius and may be effectively used in the topical therapy of canine skin infections.

고추 탄저병균의 포자 발아와 부착, 균사 생장에 미치는 화합물의 활성 검정법 확립 및 살균제의 효과 (Development of assay method for the activities of new compounds, and the effect of several fungicides against spore germination, adhesion, and myceial growth of Colletotrichum sp. causing red pepper anthracnose)

  • 김재정;김준태;박성우;박은숙;김흥태
    • 농약과학회지
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    • 제7권3호
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    • pp.159-168
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    • 2003
  • 본 실험에서는 microtiter plate를 이용하는 실내 검정법을 이용하여 고추 탄저병균의 기주체 침입 초기에 볼 수 있는 형태 변화인 포자 발아 및 부착과 균사 생장에 미치는 다양한 화합물의 활성을 대량으로 검정할 수 있는 방법을 확립하였고, 기존의 몇 가지 살균제가 가지는 고추 탄저병균의 초기 형태 분화 에 대한 억제 효과를 조사하였다. 본 실내 검정법을 표준화하기 위하여 MTT와 propanol 처리, 접종하는 포자의 농도, 배양 시간들이 병원균의 흡광도에 미치는 영향을 조사하였다. MTT를 12 시간, propanol을 1 시간 처리하였을 때, 그리고 JC24 의 포자를 $1\times10^5$개/mL의 농도로 접종하고 36 시간 배양하였을 때, 병원균의 흡광도가 일관성 있고 높게 나타났다. 살균제의 작용 특성을 알아보기 위해서 6종의 보호용 살균제, 6종의 스테롤 생합성 저해 살균제, 1종의 호흡저해제를 선발하여 microtiter plate 대량 검정 방법에서 효과를 조사하였다. 보호용 살균제 중에서 mancozeb, chlorothalonil, dithianone 등은 $6.25{\mu}g/mL$ 처리구에서도 포자의 발아, 부착과 균사의 생장을 모두 80% 이상 억제하였으나, propineb, iminoctadine, fluazinam 등은 $100{\mu}g/mL$의 처리를 2 시간만에 세척하였을 때 흡광도가 감소하는 것으로 보아 포자 부착 억제 효과는 적었다. 6종의 스테롤 생합성 저해제 중에서 hexaconazole, tebuconazole, myclobutanil 등은 포자의 발아와 균사 생육을 억제하고, 세척할 경우 JC24의 생육 억제 효과가 떨어지는 것을 보아 부착 억제 효과는 기대할 수 없었다. Nuarimol 은 $100{\mu}g/mL$로 처리한 고농도에서만 부착에 대한 억제 효과가 54.7%로 다른 억제 효과보다 우수하였으나, $6.25{\mu}g/mL$의 저농도로 처리하였을 때는 부착 억제 효과가 나타나지 않았다. Kresoxim-methyl은 6.25와 $100{\mu}g/mL$의 농도에서 포자 부착에 대한 억제율이 27.3%와 54.4%로 나타났는데, 처리하는 농도가 높아짐에 따라서 효과도 증가하였다. 이상의 결과와 같이 때 microtiter plate를 사용하여 병원균이 식물체를 침입하는 초기에 나타나는 포자 발아와 부착, 그리고 균사 생장을 검정할 수 있었으며, 본 실험 방법을 통하여 Kresoxim-methyl은 탄저병균의 포자 부착을 특이적으로 억제하는 효과가 있음을 알 수 있었다.

사미연견탕가미방(四味軟堅湯加味方)이 항암(抗癌) 및 항전이(抗轉移) 효과(效果)에 미치는 영향 (Study on Antitumor Activity of Samiyeongeontanggamibang(SYTG))

  • 배문용;강인철;김성훈
    • 대한한방종양학회지
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    • 제5권1호
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    • pp.33-46
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    • 1999
  • To evaluate the antitumor and antimetastatic effects of Samiyeongeontanggamibang(SYTG), We have examined whether SYTG can inhibit the growth of several tumor cell lines, tumor cell adhesion, experimental tumor metastasis and increase survival rate of tumor-bearing mice by inhibition of DNA topoisomrase activity. The results were obtained as follows: 1. SYTG extracts revealed an efficient cytotoxicity against A549, SK-OV-3, B16-F10, and SK-MEL-2 cell lines. 2. SYTG extracts inhibited DNA topo-isomerase I from calf thymus. 3. The T/C% in S-180 bearing ICR mice treated with SYTG was 115.8% 4. SYTG extracts significantly inhibited adhesion of A549 cell to complex extracellular matrix. 5. In pulmonary colonization assay, SYTG suppressed lung metastases in tumor cell-injected mice. 6. In CAM assay, SYTG extracts inhibited angiogenesis at $15{\mu}g/egg$ concentration as compared with control. These results suggested that SYTG extracts might be a potent inhibitor of cancer.

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페놀산의 구조가 암세포에 대한 세포독성에 미치는 영향 (Selective Cytotoxicities of Phenolic Acids in Cancer Cells)

  • 한두석;오상걸;오은상
    • Toxicological Research
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    • 제19권1호
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    • pp.45-50
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    • 2003
  • The purpose of this study was to determine the role of substituted groups in phenolic compounds to develop an anticancer agent having strong cytotoxicity against cancer cells but weak against normal cells. The phenolic compounds used in this study were gallic acid and ferulic acid with hydroxyl and carboxyl groups, syringic acid with hydroxyl, carboxyl and methoxy groups, and pyre-gallol with hydroxyl groups. Cytotoxicities of these compounds were evaluated by MTT assay for cell viability and XTT assay for cell adhesion activity in normal human skin fibroblast (Detroit 551) and human skin melanoma (SK-MEL-3) cells. Syringic acid, gallic acid and ferulic acid decreased the cell viability and cell adhesion activity in SK-MEL-3 cells but not in Detroit 551 cells while pyrogallol decreased in both cells. The susceptibility of cell viability based on the $IC_{50}$ values of MTT assay in Detroit 551 cells was in the following order: pyrogallol > gallic acid > ferulic acid > syringic acid, while it was in SK-MEL-3 cells: Syringic acid > progallol > ferulic acid > gallic acid. These results suggest that carboxyl and methoxy groups of these compounds play an important role in selectivity of cytotoxicity in normal and cancer cells.

항암활성 수종생약의 B16-Fo와 A549 암세포에 대한 항전이 효과( II ) (Antimetastatic effect of several crude drugs with antitumor activity on B16-Fo and A549 cells( II ))

  • 김성훈;유시용
    • 대한한의학회지
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    • 제17권1호
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    • pp.132-145
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    • 1996
  • Ethyl ether fractions of Ferula Resina(EFR) and Lithospermi Radix(ELR) and hexane fraction of Salviae miltiorrhizae Radix(HSR) exerted an inhibitory effect on cell adhesion of A549 to extracelluar matrix most effectively in vitro cell adhesion assay. Thus, with above fractions for the evaluation of antitumor activity, T/C% was monitored in ICR bearing S-180 and for antimetastatic effect, pulmonary colonization assay, weight of organs, changes of WBC and platelet were studied. In pulmonary colonization assay incidence rate to control was 73 % 42 %, 14 % in ELR, HSR and EFR-treated groups repsectively. Gain of lung weight was significantly inhibited in all groups while spleen weight was significantly reduced only in SMR group, but no changes in kidney and liver as compared with control. Number of platelet was significantly increased in all groups to normal range as compared with thrombocytopenic contol. WBC was significantly reduced only in LR group. These results suggest that ethyl ether fraction of Ferula Resina has more effective antimetastatic activity.

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표면특성 변화에 따른 유류분해 미생물의 토양내 거동성 조절

  • 류두현;목지예;최상일;김용미;이경애
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2003년도 추계학술발표회
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    • pp.360-362
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    • 2003
  • The adhesion of hydrocarbon degrading bacteria(HDB) differing in surface hydrophobicity was investigated. Cell wall hydrophobicity was modified chemically and physiologically. Modified adhesion deficient mutant of HDB was selected in a soil column assay Physiologically and chemical modification increased cell surface hydrophobicity. Cell surface charcteristis including BATH and zeta potential were measured. Physiological modification using ampicillin was not stable, but chemical modification was stabel. Hydrocarbon degrading potential was measured for modified and unmodifed HDB.

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유류분해 미생물의 표면특성에 따른 분해성 및 거동성 변화

  • 류두현;목지예;최명석;김진명;김동일;전경화;박소연
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2004년도 임시총회 및 추계학술발표회
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    • pp.273-276
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    • 2004
  • The adhesion of hydrocarbon degrading bacteria(HDB) differing in surface hydrophobicity was investigated. Cell wall hydrophobicity was modified chemically and physiologically. Modified adhesion deficient mutant of HDB was selected in a soil column assay. Physiologically and chemical modification increased cell surface hydrophobicity. Cell surface characteristics including BATH and FTIR were measured. Physiological modification using ampicillin was not stable, but chemical modification was stable. Hydrocarbon degrading efficiency was measured of TPH modified and unmodifed HDB.

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DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface

  • Park, Ju-Mi;Jeon, Hye-Ran;Pang, Eun-Kyoung;Kim, Myung-Rae;Kang, Na-Ra
    • Journal of Periodontal and Implant Science
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    • 제38권sup2호
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    • pp.299-308
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    • 2008
  • Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.