• Title/Summary/Keyword: Adhesion assay

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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 (활성화된 단핵구 및 대식세포의 항원제시기능에 대한 Kaempferitrin의 조절 효과)

  • Kim, Byung-Hun;Cho, Dong-Ha;Cho, Jae-Youl
    • YAKHAK HOEJI
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    • v.51 no.6
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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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    • v.14 no.11
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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
    • Journal of Veterinary Clinics
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    • v.36 no.2
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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.

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 (고추 탄저병균의 포자 발아와 부착, 균사 생장에 미치는 화합물의 활성 검정법 확립 및 살균제의 효과)

  • Kim, Jae-Jeung;Kim, Joon-Tae;Park, Sung-Woo;Park, Eun-Suk;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.7 no.3
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    • pp.159-168
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    • 2003
  • With microtiter plate, the assay method was developed for detecting the fungicidal activity of new compounds against spore germination, spore adhesion and mycelial growth of Colletotrichum sp. JC24 cal1Sing red pepper anthracnose. Also, the effects of some commercialized fungicides on fungal development like above mentioned were investigated by measuring the optical density of mycelia grown into wells of microtiter plate. For the standardization of assay method, some factors, such as the treatment of MTT and/or propanol, inodulum density and incubation period, affecting on mycelial optical density were investigated. For obtaining precise and consistent mycelial optical density, it was necessary the treatment of MTT for 12 hrs and propanol for 1 hr. inoculum density adjusted to $1\times10^5$ spores/mL and incubation period for 36 hrs at $25^{\circ}C$. For fungicidal activities, 6 protective fungicides, 6 ones inhibiting sterol biosynthesis, and one inhibiting respiration were used in this study. While mancozeb, chlorothalonil and dithianon among 6 protective fungicides inhibited strongly spore germination, adhesion, and mycelial growth at $6.25{\mu}g/mL$, propineb, iminoctadine and fluazinam inhibited intermediately spore germination and mycelial growth at $100{\mu}g/mL$. Washing above 3 fungicides with new PD broth, their activity against spore adhesion decreased. With hexaconazole, tebuconazole and myclobutanil, the tendency of the activity against fungal differentiation of the early infection stage was similar to the latter group of protective fungicides, showing the decrease of the inhibitory activity against spore adhesion by washing 2 hrs after incubation. However, kresoxim-methyl inhibited spore adhesion distinctly, depending on the applied concentrations. Based on these results, it might be able to assess the fungicidal activity of many compounds against spore germination, adhesion and mycelial growth by the use of microtiter plate in vitro. Using the assay developed in this report, it was possible to investigate the inhibitory activity of some commercialized fungicides, too.

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

  • Bae, Moon-Yong;Kang, In-Cheol;Kim, Sung-Hoon
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.5 no.1
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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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Selective Cytotoxicities of Phenolic Acids in Cancer Cells (페놀산의 구조가 암세포에 대한 세포독성에 미치는 영향)

  • 한두석;오상걸;오은상
    • Toxicological Research
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    • v.19 no.1
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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.

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

  • Kim, Sung Hoon;Ryu, Shi-Yong
    • The Journal of Korean Medicine
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    • v.17 no.1 s.31
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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
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2003.09a
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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
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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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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    • v.38 no.sup2
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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.