• Title/Summary/Keyword: 염색성 변화

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Effects of Solidago virga-aurea var. gigantea Miq. Root Extracts on the Activity and Differentiation of MC3T3- E1 Osteoblastic Cell (미역취(Solidago virga-aurea var. gigantea Miq.) 뿌리 추출물이 MC3T3-E1 조골세포의 활성과 분화에 미치는 영향)

  • Park, Jung-Hyun;Lee, Ji-Won;Kim, Hyun-Jeong;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.929-936
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    • 2005
  • The purpose of this study was to examine the ability of alkaline phosphatase (ALP) synthesis of MC3T3-E1 cells when above edible sources, Solidago virga-aurea var. gigantea Miq. root (SVR) extracts, were supplimented. MC3T3-E1 cells were cultured with $\alpha-MEM$(vehicle control), dexamethasone and genestein (positive control), and SVR extracts for 27 days. The effects of SVR MeOH extracts and its fractions on cell proliferation were measured by MTT assay. At 10, 100${\mu}g/mL$ of SVR methanol extract treated, that were elevated of cell proliferation to 140 and $120\%$ via vehicle control, respectively. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry at 3, 9, 18, and 27th day. As the results, every extracts and fractions were promoted ALP activity by time course at 1, 10, 100${\mu}g/mL$, except n-hexane and chloroform fractions. Remarkably, the MeOH extracts were increased ALP activity more than 4.4 times compared with vehicle control, 2.2 times via positive control at 27th day (p<0.05). The SVR MeOH extracts treated cells, especially at a concentration of 10${\mu}g/mL$, showed remarkably higher than vehicle-treated control cells of mineralization which were checked by Alizarin red staining. These results indicate that SVR methanol extract have an induction ability of proliferation and differentiation on osteoblast.

Expression of EGFR on the Rat Gingival Epithelia During the Experimental Tooth Movement (실험적 치아이동시 백서 치은 상피의 표피성장인자 수용체의 발현)

  • Lee, Sang-Seon;Kim, Young-Ho;Bae, Chang
    • The korean journal of orthodontics
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    • v.28 no.5 s.70
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    • pp.775-782
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    • 1998
  • The purpose of this study is to investigate the change of the EGFR mRNA expression in the rat gingival epithelium by the experimental tooth movement. We applied reciprocal force between the upper anterior teeth using NiTi open coil spring and stainless steel wire for 1, 2 3, 7 days. For the detection of EGFR mRNA, in situ hybridization was done in the tissue samples which were taken from the pressure and tension sides of teeth. The results were as follows ; 1. The expression of EGFR mRNA was increased application-time dependently. a. Day 1 mild expression on the basal and spinous cell layers b. Day 2 . moderate expression on the whole layers c. Day 3 : severe expression on the basal and spinous cell layers 4. Day 7 severe expression on the whole layers 2. The expression level of EGFR mRNA in the pressure and tension sides were similar during the whole Period of experiment except seven day application at which the cornified layer of the tension side showed moderate expression. 3. Removal of the appliance after 7-day force application lowered the level of EGRF mRNA expression. It was returned to the mild and control (rare) level at three and seven days after the removal, respectively. In conclusion, EFGR mRNA was increased by the experimental tooth movement on the rat ginigval epithelium. Up-regulation of EGFR mRNA in the gingival epithelium can be regarded as responses to the possible changes caused by the physical stersses to the oral environment to maintain the homeostatic conditions of the periodontium.

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Development of Proteomics-based Biomarkers for 4 Korean Cultivars of Sorghum Seeds (Sorghum bicolor (L.) Moench) (국내 수수 종자 분석을 위한 프로테오믹스-기반 바이오마커 개발)

  • Kim, Jin Yeong;Lee, Su Ji;Ha, Tae Joung;Park, Ki Do;Lee, Byung Won;Kim, Sang Gon;Kim, Yong Chul;Choi, In Soo;Kim, Sun Tae
    • Korean Journal of Environmental Agriculture
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    • v.32 no.1
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    • pp.48-54
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    • 2013
  • BACKGROUND: Sorghum (Sorghum bicolor (L.) Moench) ranks as the 6th most planted crop in the world behind wheat, rice, maize, soybean, and barley. The objective of this study was to identify bio-marker among sorghum cultivars using proteomics approach such as two-dimensional polyacrylamide gel electrophoresis (2-DE) coupled with mass spectrometry (MS). METHODS AND RESULTS: Proteins were extracted from sorghum seed, and separated by 2-DE. Total 652 spots were detected from 4 different sorghum seed after staining of 2-DE with colloidal Coomassie brilliant blue (CBB). Among them, 8 spots were differentially expressed and were identified using MALDI-TOF/TOF mass spectrometry. They were involved in RNA metabolism (spot1, spot 4), heat shock proteins (HSPs, spot 2), storage proteins (spot 3, spot 5, and spot 6), and redox related proteins (spot 8). Eight of these proteins were highly up-regulated in Whinchalsusu (WCS). The HSPs, Cupin family protein, and Globulin were specifically accumulated in WCS. The DEAD-box helicase was expressed in 3 cultivars except for WCS. Ribonuclease T2 and aldo-keto reductase were only expressed in 3 cultivars except for Daepung-susu (DPS). CONCLUSION(S): Functions of identified proteins were mainly involved in RNA metabolism, heat shock protein (HSP), and redox related protein. Thus, they may provide new insight into a better understanding of the charactreization between the cultivars of sorghum.

Biodisc Tissue-Engineered Using PLGA/DBP Hybrid Scaffold (DBP/PLGA 하이브리드 담체를 이용한 조직공학적 바이오 디스크 개발)

  • Ko, Youn-Kyung;Kim, Soon-Hee;Jeong, Jae-Soo;Ha, Hyun-Jung;Yoon, Sun-Jung;Rhee, John-M.;Kim, Moon-Suk;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.31 no.1
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    • pp.14-19
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    • 2007
  • Demineralized bone particle (DBP) has been used as one of the powerful inducers of bone and cartilage tissue specialization. In this study, we fabricated DBP/PLGA scaffold for tissue engineered disc regeneration. We manufactured dual-structured scaffold to compose inner cylinder and outer doughnut similar to nature disc tissue. The DBP/PLGA scaffold was characterized by porosity, wettability, and water uptake ability. We isolated and cultured nucleus pulposus (NP) and annulus fibrosus (AF) cells from rabbit intervertebral disc. We seeded NP cells into the inner core of the hybrid scaffold and AF cells into the outer portion of it. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl) -2,5- diphenyltetrazolium -bromide (MTT) test. PLGA and PLGA/DBP scaffolds were implanted in subcutaneous of athymic nude mouse to observe the formation of disc-like tissue in vivo. And then we observed change of morphology and hematoxylin and eosin (H&E). Formation of disc-like tissue was better DBP/PLGA hybrid scaffold than control. Specially, we confirmed that scaffold impregnated 20 and 40% DBP affected to proliferation of disc cell and formation of disc-like tissue.

Effect of sonicates of Treponema denticola on osteoblast differentiation (Treponema denticola 분쇄액에 의한 조골세포분화 억제효과)

  • Choi, Bong-Kyu;Kang, Jung-Hwa;Jin, Seung-Wook;Ohk, Seung-Ho;Lee, Syung-Il;Yoo, Yun-Jung
    • Journal of Periodontal and Implant Science
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    • v.33 no.1
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    • pp.79-89
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    • 2003
  • 치주질환은 세균감염에 의해 치조골이 파괴되는 염증성질환으로서 치아상실의 주된 원인이다. Treponema denticola는 성인성 치주염의 병소에서 자주 발견되는 세균으로서 부착능 및 단백분해효소생성능과 같은 독성 인자가 밝혀져 치주조직 파괴에 있어서 중요성이 강조되어 왔다. 골개조는 조골세포의 골형성및 파골세포에 의한 골흡수의 균형에 의하여 유지되며 치주염시 야기되는 치조골파괴는 조골세포 및 파골세포 기능의 불균형에 의하여 야기되는 것으로 설명되고 있다. 골세포에 대한 영향으로서 T. denticola는 파골세포의 형성을 촉진시키는 것으로 보고되었으나 조골세포에 대한 영향은 아직 밝혀져 있지 않다. 따라서 본 연구에서는 T. denticola가 골형성에 미치는 영향을 알아보고자 마우스의 두개골세포로부터 조골세포를 분리한 후 T. denticola분쇄액으로 처리하여 본 세균이 조골세포의 alkaline phosphatase(ALPase) 활성, 석회화결절 형성 및 Prostaglandin $E_2\;(PGE_2)$ 생성에 미치는 영향을 평가하였다. ALPase활성은 p-nitrophenylphosphate분해능, 석회화결절형성은 Von Kossa 염색법, 그리고 PGE2의 농도는 효소면역측정법으로 측정하였다. T. denticola분쇄액 (2.5 ug/ml)은 마우스 두개골세포의 ALPase활성을 억제하였으며 석회화결절의 형성을 감소시켰다. 또한 동일한 농도의 균분쇄액은 마우스 두개골세포의 $PGE_2$ 생산을 증가시켰다. 균분쇄액과 prostaglandin의 합성억제제인 indomethacin으로 세포를 동시에 처리한 경우 T .denticola분쇄액에 의한 $PGE_2$의 생산은 감소되었으나, ALPase의 활성억제에는 변화가 없었다. 균분쇄액을 열처리하여 마우스 두개골세포에 처리하였을 때에도 ALPase의 활성이 억제되는 것에는 변함이 없었다. 이러한 결과는 T. denticola의 구성성분 중 열에 안정한 물질이 prostaglandin과 무관한 경로를 통해 조골세포의 분화를 억제함을 시사하며 이와 같은 T. denticola에 의한 골형성억제가 치주염시 야기되는 치조골 파괴에 관여할 수 있을 것으로 생각된다.

A simple screening method using lignoceullulose biodegradation for selecting effective breeding strains in Agaricus bisporus (리그노셀룰로오스 생물학적 분해를 이용한 간단한 양송이 육종효율 우수 균주 선발)

  • Oh, Youn-Lee;Nam, Youn-Keol;Jang, Kab-Yeul;Kong, Won-Sik;Oh, Min ji;Im, Ji-Hoon
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.134-138
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    • 2017
  • The white button mushroom, Agaricus bisporus, is commercially the fifth most important edible mushroom, accounting for the production of 9,732 tons of mushrooms in Korea in 2015. The genus Agaricus has been known for its potential to degrade lignocellulosic materials. Chemical analyses carried out during the cultivation of A. bisporus indicated that the cellulose, hemicellulose, and lignin fractions were changed preferentially for both vegetative growth and sexual reproduction. We screened A. bisporus strains for effective biodegradation through extracellular enzyme activity using cellulase, xylanase, and ligninolytic enzymes. The enzyme biodegradations were conducted as follows: mycelia of collected strains were incubated in 0.5% CMC-MMP (malt-mops-peptone), 0.5 Xylan-MMP, and 0.5% lignin-MMP media for 14 days. Incubated mycelia were stained with 0.2% trypan blue. Eighteen strains were divided into 8 groups based on different extracellular enzyme activity in MMP media. These strains were then incubated in sterilized compost and compost media for 20 days to identify correlations between mycelial growth in compost media and extracellular enzyme activity. In this study, the coefficient of determination was the highest between mycelial growth in compost media and ligninolytic enzyme activity. It is suggested that comparison with ligninolytic enzyme activity of the tested strains is a simple method of screening for rapid mycelial growth in compost to select good mother strains for the breeding of A. bisporus.

Comparison of Neural Cell Differentiation of Human Adipose Mesenchymal Stem Cells Derived from Young and Old Ages (연령별 지방 중간엽 유래 줄기세포의 신경세포로의 분화 능력 비교)

  • Jo, Jung-Youn;Kang, Sung-Keun;Choi, In-Su;Ra, Jeong-Chan
    • Development and Reproduction
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    • v.13 no.4
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    • pp.227-237
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    • 2009
  • Recently, adipose mesenchymal stem cells (AdMSC) that are similar to bone marrow MSC and blood derived MSC are thought to be another source for stem cell therapy. However, the diseases that can be applied for stem cells therapy are age-dependent degenerative diseases. Accordingly, the present study investigated the growth and differentiation potential to neural cells of human AdMSC (hAdMSC) obtained from aged thirty, forty and fifty. The growth of cells and cell viability were measured by passage and neural differentiation of hAdMSC was induced in neural differentiation condition for 10 days. Our results demonstrated that cell number, viability and morphology were not different from hAdMSC by age and passage. Immunofluorescence analysis of neural cell marker (TuJ1, NSE, Sox2, GFAP or MAP2) demonstrated no significant differences in neural cell differentiation by age and passage. As the number of passage was increased, the mRNA level of MAP2 and Sox2 was decreased in hAdMSC from age of 50 compared to hAdMSC from age of 30. In conclusion, the present study demonstrated that ability of neural cell differentiation of hAdMSC was maintained with ages, suggesting that autologous stem cells from aged people can be applied for stem cell therapy with age-dependent neural disease with the same stem cell quality and ability as stem cell derived from young age.

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Toxoplasmacidal Effect of HL-60 Cells Differentiated by Dimethylsulfoxide (Dimethylsulfoxide로 분화시킨 HL-60 세포의 yoxoplasma 파괴 효과)

  • Choe, Won-Yeong;Nam, Ho-U;Yu, Jae-Eul
    • Parasites, Hosts and Diseases
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    • v.26 no.4
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    • pp.229-238
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    • 1988
  • In vitro culture of Toxoplasma gondii in HL-60 cells and cell-mediates immunity against Toxoplasma in dimethylsulfoxide(DMSO) -induced HL-60 cells, i.e., differentiation into granulocytes, were pursued. HL-60 calls were treated with various concentrations of DMSO, and 1.3%(v/v) for 3 day incubation was chosen as the optimal condition icy differentiation into granulocytes. The degree of differentiation was assayed in physiological and functional aspects in addition to morphological point. When treated with 1.3% DMSO for 3 days, HL-60 cells did not synthesiar DNA materials beyond background level, and showed active chemotactic response to chemotactic peptide, formal-methionyl-leucyl-phenylalanine(FMLP). Morphologically promyelocytes of high nuclearlcytoplasmic(NIC) ratio changed to granulocytes of relatively low WJC ratio. The relationships between HL-60 cells or DMSO-induced HL-60 cells and Toxoplasma were examined after stain with Giemsa and Buorescent dye (acridine orange). HL-60 cells did not show any sign of torso- plasmacidal activity but showed intracellular proliferation of Texoplasma to form rosette for 72 hr co-culture. In contrast, OMSO-induced HL-60 cells phagocytosed Toxoplasma within 1 hr, and performed a process of intracellular digestion of Toxoplasma thereafter. With the above results, it is suggested that phagosome-Iysosome fusion is one of the critical events for the parasitism by Toxoplasma or for susceptibility of host cells. The in vitro culture system of this study has offered a defined condition to study the protozoan parasite-host cell interactions.

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STUDIES OF GINSENG EXTRACT ON AGE-RELATED ENZYMES (고려인삼 추출물이 노화관련효소에 미치는 영향)

  • Cho Young Dong;Koo Bon Sook;Lee Song Jae
    • Proceedings of the Ginseng society Conference
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    • 1984.09a
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    • pp.37-43
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    • 1984
  • 노화는 모든 다세포 유기물의 특징이다. 노화가 됨에 따라서 효소활성 및 면역반응의 감소와 과산화지질과 지방갈색물질의 축적, 효소와 염색질을 포함하는 단백질 구조의 변화, 호르몬계의 불균형 등이 일어난다. 그렇지만, 노화가 어떻게 일어나는지에 관하여는 현재까지 확실하지 않다. 본 연구진은 노화와 관련된 효소들에 관하여 연구를 하여 왔으며, 노화가 진행되는 동안의 효소의 활성을 유지시켜주거나, 또는 효소의 활성이 감소되는 것을 지연시켜 주는 물질을 찾고자 노력하였다. 그 가운데 하나로서, 고려인삼 추출물을 흰쥐에 기간별로 투여하여 효소활성의 차이, 열에 대한 안정성, 기질에 대한 친화력, 전기 영동 상의 이동성과 면역적인 반응을 대조군과 비교하였다. Glucose-6-phosphate dehydrogenase, 6-phosphog-luconate dehydrogenase, glutathione redutease, glutathion peroxidase와 같은 노화와 관련된 효소들의 활성을 고려인삼 추출물을 1개월간 흰쥐에 ($60{\~}80$g)투여하여 대조군과 비교 조사하였으나, 별 차이가 없었다. 그러나 고려인삼 추출물을 15개월간 투여하였을 때에는 이러한 노화관련 효소들의 활성이 급격히 증가함이 ($70{\~}200\%$) 관찰되었다. 예견된 바와 같이, 효소의 열에 대한 안정성과 기질에 대한 친화력도 증가함이 관찰되었다. 그러나 glucose-6-phosphate dehydrogenase의 경우에서 전기영동상의 차이 및 면역적인 반응은 대조군과 유사하였다. 이상의 결과는 고려인삼 추출물이 노화와 관련된 효소들의 활성이 감소되는 것을 지연시켜줄 수 있으며, 노화를 어느정도 지연시켜 줄 수 있음을 의미한다. 이와 같은 결과를 포함한 실험자료를 국제 인삼심포지움에서 발표할 것이다.$-tocopherol의 항 산화작용을 더욱 효과적으로 촉진 시킬 것으로 생각된다.-L(독성 호르몬-L)의 작용을 억제함으로서 암환자의 체중 감소를 방지하고, 식욕감퇴를 개선할것으로 생각된다.해되었으며, $Rb_{1}$은 장내의 효소와 tetracycline-resistantant bacteria에 의해 Rd와 2 종류의 미확인 물질로 분해되었다.xA_{2}$ synthetase 억제제인 imidazole의 효과와 유사하였다. 4. G-Re는 $1{\times}10^{-5}g/ml$ 이하의 농도에서는 효과가 없으나 $1{\times}10^{-4}g/ml$ 이상의 농도에서 농도의존적으로 유의성 있는 $PGE_{2},\;PGF_{2}{\alpha},\;TXB_{2}$의 생성억제와 함께 6-keto-$PGF_{1}{\alpha}$ 증가를 보였다. 이는 prostacyclin synthetase를 자극하는 serotonin의 효과와 같은 작용으로서 prostacyclin synthetase 억제제인 tranylcypromine에 대하여 길항효과를 보였다. 5. $TxB_{2}$생성억제 작용을 나타내는 ginsenoside들의 효과를 뒷받침하기 위하여 인삼 saponin 성분을 전처치한 patelet rich plasma에서 혈소판 응집시험 결과, ADP로 유도된 혈소판 응집반응에는 모든 인삼 saponin 성분들이 효과가 없었으나 arachidonic acid로 유도된 혈소판 응집반응에는 $G-Rb_{2}$, G-Rc, G-Re의 순으로 농도 의존적인 억제현상을 보였다. 이상의 결과와 같이 인삼 saponin 성분들은 arachidonic acid로부터 cyclooxygenase를

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Relationship between the Regulator of Calcineurin 1-4 Isoform and In Vitro Osteoclast Differentiation (Regulator of calcineurin 1-4과 파골세포 분화의 관련성)

  • Park, Kyeong-Lok
    • Journal of Life Science
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    • v.25 no.2
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    • pp.223-230
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    • 2015
  • Regulator of calcineurin 1 (RCAN1) is an endogenous calcineurin inhibitor that plays an important role in the pathogenesis of diseases related to the calcineurin-NFATc1 signaling pathway. The RCAN1-4 isoform is subject to NFATc1-dependent regulation. During receptor activator of nuclear factor kappa-B ligand (RANKL)-stimulated osteoclastogenesis, the calcineurin-NFATc1 pathway is critical. Because there is little information available on the role of RCAN1 in osteoclast differentiation, this study investigated whether changes in RCAN1 expression are related to the calcineurin-NFATc1 pathway and osteoclast differentiation. Mouse bone marrow monocytes (BMMs) were treated with 50 ng/ml of RANKL and M-CSF. Expression levels of NFATc1, calcineurin, and RCAN1 isoforms were determined using RT-PCR and Western blotting. Osteoclast differentiation was examined using tartrate-resistent acid phosphatase (TRAP) staining. To evaluate the effect of RCAN1 overexpression on osteoclastogenesis, cells were transfected with a mouse RCAN1-4 cDNA plasmid. After RANKL stimulation of BMMs, expression of NFATc1 and RCAN1 was increased at the mRNA and protein level, while calcineurin expression was unchanged. When the RCAN1-4 gene construct was transfected, the expression of RCAN1 protein was not increased despite several-fold increases in RCAN1-4 mRNA expression. Regardless of RANKL stimulation, over-expression of RCAN1-4 tended to reduce NFATc1 expression and knock-down of RCAN1 increase it. While BMMs transfected with the RCAN1-4 vector were differentiated into distinct osteoclasts, their phenotypes did not vary from those of mock controls. These results suggest that RCAN1 has a limited effect on the calcineurin-NFATc1 pathway during RANKL-stimulated osteoclast differentiation.