• 제목/요약/키워드: Cell-based assay

검색결과 667건 처리시간 0.026초

삼황세제가감방(三黃洗劑加減方)의 항균, 항염 및 항알레르기 효과에 대한 실험적 연구 (A Study on the Anti-microbacterial Activity, Anti-inflammatory and Anti-allergic Effects of Samhwangseje gagambang(SHB))

  • 원영호;심은기;안찬근;박민철;황충연
    • 한방안이비인후피부과학회지
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    • 제18권1호
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    • pp.135-153
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    • 2005
  • Herbal mixture water extract of (Phellodendron amurense, Scuellaria baiklensis, Spphora flavescens, Lithospermum erythrorhizon, Mellaphis chinesis, Alumite, Zanthoxylum schinifolium, Glycyrrhiza uralensis), which exhibit several beneficial effects including acne and skin diseases, was tested for anti-microbial activity and anti-inflammation effects. The herbal mixture extract showed antimicrobial activity against Stapylococcus epidermis, Propionibacterium acne, and Malassezia furfur. The growth of Stapylococcus epidermis, Propionibacterium acne, and Malassezia furfur was inhibited allergy and LPS induced cyclooxygenase-2(COX-2)gene expression in RAW24.7macrophage. The results indicated th ear swelling and histamine release induced by compound 48/80 were dose-dependently reduced, ranging 11-38% and 11-56%, respectively. Furthermore the extract inhibited the expression of LPS-induced COX-2 proteins and mRNAs without an appreciable cytotoxic effects on RAW264.7 cells. The cytotoxicity of the extract using M7T assay showed the cytotoxicity of 7 and 18% against L929 cell line. Based on these results, it is concluded that the herbal mixture water extract can be applied to the acne and skin diseases therapy.

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대황(大黃), 황금(黃芩), 황백(黃柏) 복합 추출물의 항균, 항염 및 항알레르기 효과에 대한 실험적 연구 (A Study on the Anti-microbacterial Activity, Anti-inflammatory and Anti-allergic Effects of mixture of Rheum coreanum Scutellaria baikalensis Phellodenron amurense)

  • 손대범;송성필;황치환;홍석훈;황충연
    • 한방안이비인후피부과학회지
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    • 제18권1호
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    • pp.154-171
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    • 2005
  • Herbal mixture water extract of (Rheum coreanum Scutellaria baikalensis, Phellodendron amurese), which exhibit several beneficial effects including acne and skin diseases, was tested for anti-microbial activity and anti-inflammation effects. The herbal mixture extract showed antimicrobial activity against Stapylococcus epidermis and Propionbacterium acne. The growth of Stapylococcus epidermis and Propionibacterium acne was inhibited completely by addition of 1.0% of the extract. Also in the present study we examined the mixture extract on compound 48/80 induced allergy and LPS induced cyclooxygenase-2(COX-2) gene expression in RAW264.7 macrophage. The results indicated the ear swelling and histamine release induced by compound 48/80 were dose-dependently reduced, ranging 18-36% and 10-61%, respectively. Furthermore the extract inhibited the expression of LPS-induced COX-2 proteins and mRNAs without an appreciable cytotoxic effects on RAW264.1 cells. The cytotoxicity of the extract using MTT assay showed the cytotoxicity of 6% and 13% against L929 cell line. Based on these results, it is concluded that the herbal mixture water extract can be applied to this acne and skin diseases therapy.

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Instrumentation and Software for Analysis of Arabidopsis Circadian Leaf Movement

  • Kim, Jeong-Sik;Nam, Hong-Gil
    • Interdisciplinary Bio Central
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    • 제1권1호
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    • pp.5.1-5.4
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    • 2009
  • This article is an addendum to the authors’ previous article (Kim, J. et al. (2008) Plant Cell 20, 307-319). The instrumentation and software described in this article were used to analyze the circadian leaf movement in the previous article. Here, we provide detailed and practical information on the instrumentation and the software. The source code of the LMA program is freely available from the authors. The circadian clock regulates a wide range of cyclic physiological responses with a 24 hour period in most organisms. Rhythmic leaf movement in plants is a typical robust manifestation of rhythms controlled by the circadian clock and has been used to monitor endogenous circadian clock activity. Here, we introduce a relatively easy, inexpensive, and simple approach for measuring leaf movement circadian rhythms using a USB-based web camera, public domain software and a Leaf Movement Assay (LMA) program. The LMA program is a semi-automated tool that enables the user to measure leaf lengths of individual Arabidopsis seedlings from a set of time-series images and generates a wave-form output for leaf rhythm. This is a useful and convenient tool for monitoring the status of a plant's circadian clock without an expensive commercial instrumentation and software.

Cytochrome P-450 3A4 proximal promoter activity by histone deacetylase inhibitor in HepG2 cell.

  • Kim, Ja-Young;Ahn, Mee-Ryung;Sheen, Yhun-Yhong
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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    • pp.88-88
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    • 2003
  • Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to +64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

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Folate-Targeted Nanostructured Lipid Carriers (NLCs) Enhance (Letrozol) Efficacy in MCF-7 Breast Cancer Cells

  • Sabzichi, Mehdi;Mohammadian, Jamal;Khosroushahi, Ahmad Yari;Bazzaz, Roya;Hamishehkar, Hamed
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권12호
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    • pp.5185-5188
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    • 2016
  • Objective: Targeted-drug-delivery based lipid nanoparticles has emerged as a new and effective approach in cancer chemotherapy. Here, we investigated the ability of folate-modified nanostructured lipid carriers (NLCs) to enhance letrozol (LTZ) efficacy in MCF-7 breast cancer cells. Methods: New formulations were evaluated regarding to particle size and scanning electron microscope (SEM) features. Anti-proliferative effects of LTZ loaded nanoparticles were examined by MTT assay. To understand molecular mechanisms of apoptosis and cell cycle progression, flow cytometric assays were applied. Results: Optimum size of nanoparticles was obtained in mean average of $98{\pm}7nm$ with a poly dispersity index (PDI) of 0.165. The IC50 value was achieved for LTZ was $2.2{\pm}0.2{\mu}M$. Folate-NLC-LTZ increased the percentage of apoptotic cells from 24.6% to 42.2% compared LTZ alone (p<0.05). Furthermore, LTZ loaded folate targeted NLCs caused marked accumulation of cells in the subG1 phase. Conclusion: Taken together, our results concluded that folate targeted LTZ can be considered as potential delivery system which may overcome limitations of clinical application of LTZ and improve drug efficacy in tumor tissue.

Increased Association of ${\alpha}$-synuclein to Perturbed Cellular Membranes

  • Kim, Yoon-Suk;Lee, Seung-Jae
    • 대한의생명과학회지
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    • 제17권2호
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    • pp.167-171
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    • 2011
  • [ ${\alpha}$ ]synuclein (${\alpha}$-syn) is implicated in the pathogenesis of Parkinson's disease (PD) and other related diseases. We have previously reported that ${\alpha}$-syn binds to the cell membranes in a transient and reversible manner. However, little is known about the physiologic function and/or consequence of this association. Here, we examined whether chemically induced perturbations to the cellular membranes enhance the binding of ${\alpha}$-syn, based on hypothesis that ${\alpha}$-syn may play a role in maintenance of membrane integrity or repair. We induced membrane perturbations or alterations in ${\alpha}$-syn-overexpressing human neuroblastoma cells (SH-SY5Y) by treating the cells with hydrogen peroxide ($H_2O_2$) or oleic acid. In addition, membranes fractionated from these cells were perturbed by treating them with proteinase K or chloroform. Dynamic interaction of ${\alpha}$-syn to the membranes was analyzed by the chemical cross-linking assay that we developed in the previous study. We found that membrane interaction of ${\alpha}$-syn was increased upon treatment with membrane-perturbing reagents in a dose and time dependent manner. These results suggest that perturbations in the cellular membranes cause increased binding of ${\alpha}$-syn, and this may have significant implication in the physiological function of ${\alpha}$-syn in cells.

Cytochrome P-450 3A4 proximal promoter activity by histone deacetylase inhibitor in HepG2 cell.

  • Kim, Ja-Young;Ahn, Mee-Ryung;Sheen, Yhun-Yhong
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 추계국제학술대회
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    • pp.178-178
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    • 2003
  • Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this Is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to +64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

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돼지분변에서 PCR에 의한 Lawsonia intracellularis 검색 (Detection of Lawsonia intracellularis in swine feces by polymerase chain reaction)

  • 장성준;김정화;김영태;김기향;김중규;김영욱;최일영
    • 한국동물위생학회지
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    • 제24권1호
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    • pp.43-50
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    • 2001
  • Swine proliferative enteritis(SPE) caused by inかsoma intracellularis is a common enteric disaese of grower and finisher pig. Swine affected with SPE show variable clinical signs including diarrhea, weight loss, aberrant growth and death. The characteristic lesion of ileitis at necropsy is marked thickening of the last section of the small intestine. The inner lining of the thickened intestine proliferates almost like a cancer and curved rod bacteria(L intracellularis) are always seen inside the intestinal wall. Infected swine shed the organism in the feces. Isolation and growth of pure L intracellularis in vitro requires a suitable cell culture. This procedure is difficult and not a practical means of diagnosis, thus the polymerase chain reaction(PCR) test of feces can be used to determine whether a pig is shedding the infective organism. A sensitive assay based on amplification of a 319bp DffA fragment of the L intracellularis of Swine proliferative enteritis was attempted for the detection of the organism in the 62 feces of swine. L intracellularis was identified on three herds and detected in 6 fecal samples, representing a infection rate of 9.7%. The PCR was very sensitive and specific on the individual level. The PCR technique could be very useful for the diagnosis of this disease.

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Microbial Detection and Identification Using Biosensors

  • Kim, Sol
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2008년도 International Meeting of the Microbiological Society of Korea
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    • pp.135-135
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    • 2008
  • Various biosensors were evaluated for identifying and detecting foodborne pathogens in a rapid and effective manner. First, five strains of Escherichia coli and six strains of Salmonella were identified using Fourier transform infrared spectroscopy and a statistical program. For doing this, lipopolysaccharides (LPSs) and outer membrane proteins (OMPs) were extracted from a cell wall of each bacterial strain. As a result, each strain was identifed at the level of 97% for E. coli and 100% for Salmonella. Second, E. coli O157:H7, S. Enteritidis, and Listeria monocytogenes were identified by multiplex PCR products from four specific genes of each bacteria using a capillary electrophoresis (CE). Also, ground beef for E. coli O157:H7, lettuce for S. Enteritidis, and hot dog for L. monocytogenes were used to determine the possibility of detecting pathogens in foods. Foods inoculated with respective pathogen were cultivated for six hours and multiplex PCR products were obtained and assessed. The minimum detection levels of tested bacteria were <10 cells/g, <10 cells/g, and $10^4$ cells/g for E. coli O157:H7, S. Enteritidis, and L. monocytogenes, respectively. Third, it was possible to detect S. Typhimurium in a pure culture and lettuce by a bioluminescence-based detection assay using both recombinant bacteriophage P22::luxI and a bioluminescent bioreporter. In addition, bacteriophage T4 was quantitatively monitored using E. coli including luxCDABE genes.

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A Membrane-Array Method to Detect Specific Human Intestinal Bacteria in Fecal Samples Using Reverse Transcriptase-PCR and Chemiluminescence

  • KIM PYOUNG IL;ERICKSON BRUCE D;CERNIGLIA CARL E.
    • Journal of Microbiology and Biotechnology
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    • 제15권2호
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    • pp.310-320
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    • 2005
  • A membrane-based oligonucleotide array was used to detect predominant bacterial species in human fecal samples. Digoxygenin-labeled 16S rDNA probes were generated by PCR from DNA that had been extracted from fecal samples or slurries. These probes were hybridized to an array of 120 oligonucleotides with sequences specific for 40 different bacterial species commonly found in human feces, followed by color development using an alkaline phosphatase-conjugated antibody and NBT /BCIP. Twenty of the species were detected by this method, but E. coli, which was present at $\~$1 $\times 10$^5$ CFU per gram feces, was not detected. To improve the sensitivity of this assay, reverse transcriptase-PCR was used to generate probes from RNA extracted from fecal cultures. Coupled with a chemiluminescence detection method, this approach lowered the detection limit for E. coli from $\~1$ $\times 10$^6$ to ${\leq}$ 1 $\times 10$^5$ These results indicate that the membrane-array method with reverse transcriptase-PCR and chemiluminescence detection can simultaneously identify bacterial species present in fecal samples at cell concentrations as low as${\leq}$ 1 $\times 10$^5$ CFU per gram.