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- 저자 : ( Sung Mee Lim ) (검색결과 298 건)
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12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할
임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 忠南大學校 癌共同硏究所 1998 癌共同硏究所 硏究誌 Vol.2 No.1
Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_(2) at 37°C. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KC1, 60 mM HEPES, pH 7.9, 25 mM MgCl_(2), 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2μg of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.
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12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할
임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-
Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_2 at 37℃. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KCI, 60 mM HEPES, pH 7.9, 25 mM MgCI_2, 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2 ㎍ of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.
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정자림,김미경,권상호,김미정 대구효성가톨릭대학교 식품과학연구소 1993 식품과학지 Vol.5 No.-
건대추를 이용한 소금절임과정중 품질변화를 조사한 결과는 다음과 같다. 절임중의 증량변화는 온도와 염도의 차이에 따라 상당한 차이를 나타내었으며 건대추중량의 260-280%이상의 증가현상이 나타날 경우는 조직이 흐트러지고 심한 연화현상을 동반하였다. 특히 온도가 30℃로 높을 경우에 이러한 현상이 두드러지게 나타났다. 절임온도가 10℃일때 그리고 염농도가 10%일때가 바람직하였다. 절임액으로의 물질의 용출정도는 염도가 높고 온도가 높을 수록 거의 비례적으로 높았다. 염절임 대추의 색상은 온도 10℃, 염농도 10%일때가 좋았으며 온도가 높고 염농도가 이보다 높을 경우는 색상이 불량하였다. Quality changes during salting of dried jujube fruits were investigated. The summarized results were as follows. The changes in the weight of the jujube fruit showed great difference according to the difference of temperature and salt concentration during salting. Softening and decay of tissue appeared when the weight of dried jujube increased over 260-280% during salting. Especially, the phenomena was serious at above 30℃. The most disirable salt concentration and temperature for jujube salting were estimated by color and tissue state, and best condition was 10% and 10℃, respectively. Elution amounts of jujube substance from the fruit to salt solution during salting became high when the salt concentration and temperature were high during salting.
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김대성,노성택,이장천,임규상,신미란,우원홍,문연자 한국전통의학연구소 2006 한국전통의학지 Vol.15 No.1
The aim of this study was to investigate the effect of ethanol extract of Fagopyrum escuentum(FE) on the melanogenesis. To determine whether ethanol extract of FE suppress melanin synthesis in cellular level, B16F10 melanoma cells were cultured in the presence of different concentrations of FE ethanol extract. In the present study, the author examined the effects of FE ethanol extract on cell proliferation, melanin contents, tyrosinase activity. Cell proliferation was slightly increased by treatment with ethanol extract of FE (25-200 ㎍/ml). The ethanol extract of FE effectively suppressed melanin contents at a dose of 100 ㎍/ml. It was observed that the color of cell pellets was totally whitened compared with the control. The ethanol extract of FE inhibited tyrosinase activity, regulate melanin biosynthesis as the key enzyme in melanogenesis. These results suggest that the ethanol extract of FE exerts its depigmenting effects through the suppression of tyrosinase activity. And it may be a potent depigmetation agent in hyperpigmentation condition.
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사람 태반조직 DNA Topoisomerase Ⅰ에 대한 Phosphorylation의 영향
곽상태,김승민,박종일,손미영,임규,황병두 충남대학교 의과대학 지역사회의학연구소 1996 충남의대잡지 Vol.23 No.2
DNA topoisomerase I was prepared from human term placenta and the effects of phosphorylation/dephosphorylation to the enzyme activity have been investigated. hosphorylation of the enzyme by protein kinase'A or casein kinase II did not affect the activity of DNA topoisomerase I. But dephosphorylation of the enzyme by calf intestinal alkaline phosphatase decreased the DNA topoisomerase I activity. The inhibitory effects of camptothecin or 10-OH-camptothecin on the enzyme were not related to dephosphorylation of the enzyme. These results suggest that DNA topoisomerase I from human term placenta present in forms of phosphorylated, and regulation of the enzyme activity is related with phosphorylation/dephosphorylation of the enzyme.
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김순동,임영숙,조성경,김미향 대구효성가톨릭대학교 식품과학연구소 1992 식품과학지 Vol.4 No.-
감의 연화에 따른 물성 변화와 이에 따른 세포벽 다당류의 조성과 함량 및 column chromatography를 통한 계략적인 분자량의 변화를 조사하였다. 조세포벽의 함량은 연화된 감에서 현저히 감소되었다. 또한 온도변화에 따른 감의 물성변화와 세포벽 다당류의 변화를 조사하였다. 그 결과 25℃에서 가장 쉽게 연화 되었으며 45℃에서는 연화가 억제되었다. 연화가 진행되는 동안에 부착성은 증가하였으나 경도는 감소되었다. 이러한 현상은 25℃에서 가장 현저하게 나타났다. 조세포벽의 함량은 연화된 감에서 현저히 감소되었은데 구체적으로 ionically associated pectin(IAP)은 59%, covalently bounded pectin(CBP)은 60%, hemicellulose fraction 2(HF2)는 74%가 감소된 반면 hemicellulose fraction 1(HF1)과 cellulose fraction(CF)은 큰 변화가 없었다. 연화된 감의 IAP, CBP에서는 pentose와 hexose의 감소율이 높았다. HF1과 HF2역시 uronic acid의 함유율이 높았는데 HF2가 현저하였다. 또 연화에 따라 HF1은 pentose와 HF2는 uronic acid와 hexose의 손실율이 높았다. 특히 HF2는 수백만 단위의 거대분자가 만단위까지 저분자화 하였다. 이상의 결과에서의 감의 연화는 CBP에 연결된 hemicellulose중 특히 HF2의 저분자화와 밀접한 관련이 있는 것으로 판단 되었다. The changes of cell wall polysaccharides were studies by gel filteration chromatography. Various cell wall polysaccharides such as ionically associated pectin(IAP), covalently bounded pectin(CBP), 2N potasium hydroxide soluble hemicellulosic fraction(HF1) and 0-3N potasium hydroxide soluble hemicellulosic fraction(HF2) were fractionated by chemical method. The content of crude cell wall remarkably decreased in the soft persimmon. The changes of texture, composition of cell wall polysaccharides during preserving of persimmon at various temperature were estimated to investigate the softening characteristics of persimmon fruit. The softening of persimmon was the most promoted at 25℃, Whereas it was inhibited at 45℃. During softening adhesiveness increased and hardness decreased. This phenomenon was obvious at 25℃. The decreasing rates of IAP, CBP and HF2 were 59, 60 and 74%, respectively, while HF1 and cellulose changed only a little during softening. The loss rate of pentose in IAP, of hexose in CBP, of hexose and uronic acid in HF2, of pentose in HF1 increased during softening. Though apparent molecular weight of all polysaccharides shifted from high molecular weight to low molecular weight polymer, the shifting degree of CBP and HF2 was especially remarkable during softening. It is suggested that the servere softening phenomenon of persimmon involved the degradation and dissolution of wall bounded-CBP and HF2 which were associated with each other.
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수산화칼슘 처리된 Porphyromonas endodontalis Lipopolysaccharide가 다형핵백혈구의 IL-1과 TNF-α 생성에 미치는 영향에 관한 연구
박찬제,박동성,유현미,오태석,임성삼 大韓齒科保存學會 2002 Restorative Dentistry & Endodontics Vol.27 No.5
Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines , namely, interleukin 1 and tumor necrosis factor-α from immune cells, Although monocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-α. Calcium hydroxide has been shown to be an effective medicament in root canal infectionsm, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)_2 may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromanas endodonatalis LPS with calcium hydroxide alters its biological action as neasured by human PMN secretion of IL-1 and TNF-α, and it was compared with Escherichia coli LPS. P.endodonatalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified . 100 ㎍/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)_2 at 37℃ for 7 days. The supernatants were subjected to ultrafiltration. And the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4 × 10^6 cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10㎍/ml) for 24 hours at 37℃ in 5% CO_2 incubator. The supernatants of cells were collected and the levels of IL-1α, IL-1β and TNF-α were measured by enzyme-linked immunosorbent assay. The results were as follows; 1. The levels of IL-1α, IL-1β, TNF-α from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05). 2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05). while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05). 3. The levels of secretion or all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05). 4. The levels of all three cytokines released from PMN stimulated with P.endodontalis LPS were significantly lower than those released from PMN stimulated with E. coli LPS (p<0.05).
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