• Title/Summary/Keyword: D-ra

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Enhancement of anti-inflammatory and anti-tumorigenic properties of 3D-spheroid formed mesenchymal stem cells derived from rheumatoid arthritis joints

  • Seung-Chan Lee;Chae-Yeon Hong;Yong-Ho Choe;Tae-Seok Kim;Won-Jae Lee;Gyu-Jin Rho;Sung-Lim Lee
    • 한국동물생명공학회지
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    • 제37권4호
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    • pp.246-254
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    • 2022
  • Current studies have revealed the capacity of mesenchymal stem cells (MSCs) in term of immunomodulatory properties, and this distinct potential is downgraded according to the disease duration of patients-derived MSCs. In order to enhance the immunomodulatory and anti-tumorigenic properties of the rheumatoid arthritis (RA) joints-derived MSCs, we aggregate synovial fluid-derived MSCs from RA joints (RA-hMSCs) into 3D-spheroids by the use of hanging drop culture method. Cells were isolated from synovial fluids of RA joints with longstanding active status over 13 years. For aggregation of RA-hMSCs into 3D-spheroids, cells were plated in hanging drops in 30 μL of advanced DMEM (ADMEM) containing 25,000-30,000 cells/drop and cultured for 48 h. To analyze the comparative immunomodulatory effects of 3D-spheroid and 2D monolayer cultured RA-hMSCs and then cells were cultured in ADMEM supplemented with 20% of synovial fluids of RA patients for 48 h and were evaluated by qRT-PCR for their expression of mRNA levels of inflammatory and anti-inflammatory markers. Cellular aggregation of RA-hMSCs was observed and cells were aggregate into a single sphere. Following treatment of RA patient's synovial fluids into the RA-hMSCs, spheroids formed RA-hMSCs showed significantly (p < 0.05) higher expression of TNFα stimulated gene/protein 6 (TSG-6) than the monolayer cultured RA-hMSCs. Therefore, the 3D-spheroid culture methods of RA-hMSCs were more effective than 2D monolayer cultures in suppressing inflammatory response treated with 20% of RA-synovial fluids by expression of TNFα (TSG-6) according to the immune response and enhanced secretion of inflammatory factors.

인체 유방암 세포에서 retinoids의 영향에 대한 연구 (Effect of Retinoids on Human Breast Cancer Cells)

  • 윤현정;신윤용;공구
    • 한국환경성돌연변이발암원학회지
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    • 제24권2호
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    • pp.51-66
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    • 2004
  • Retinoids, better known as vitamin A, have been reported to inhibit the growth of several breast cancer cell lines in culture and to reduce breast tumor growth in animal models. Furthermore, retinoids can augment the action of other breast cancer cell growth inhibitors both in vitro and in vivo. Clinically, interest has increased in the potential use of retinoids for the prevention and treatment of human breast cancer. We have examine the effect of all-trans retinoic acid(tRA) and 9-cis retinoic acid(9-cis RA) on human breast cancer cell(MCF-10A, T47-D, MCF-7) proliferation using MTT assay and cell cycle analysis(FACS). Overexpression of cyclin D1 protein is observed in the majority of breast cancers, suggesting that dysregulated expression of cyclin D1 might be a critical event in breast cancer carcinogenesis. We investigated whether tRA and 9-cis RA might affect expression of cyclin D1 on human breast cancer cells(MCF-10A, T47-D, MCF-7) using RT-PCR and west-ern bolt. In MCF-10A cells, either tRA or 9-cis RA treatment did not affect the cell proliferation. In T47-D cells and MCF-7 cells, either tRA or 9-cis RA treatment showed the inhibition of the cell proliferation over control cells and also inhibit the estrogen stimulated cell proliferation when it was given together with estrogen. The effect of retinoids was dose- and time- dependent. T47-D cells treated with 1.0 $\muM$ tRA undergo G0/G1-phase arrest by Day 5. MCF-7 cells treated with 1.0 $\muM$ tRA undergo S-phase arrest by Day 5. All-trans retinoic acid(tRA) and 9-cis retinoic acid(9-cis RA) inhibited the cyelin D1 mRNA and protein expression levels of human MCF-7 and T47-D breast carcinoma cells in vitro. The data indicate that retinoids can reduce cyclin D1 expression levels in a variety of breast cell lines in vitro and result in inhibition of cell proliferation. tRA-mediated growth inhibition and cyclin D1 expression inhibition is more potent than 9-cis RA mediated that. tRA-mediated inhibition effect is more potent on T47-D cells than on MCF-7 cells. Our data suggest that retinoids activity is different according to property of cell lines. Future chemoprevention of breast cancer studies using retinoids will be necessary to determine the mechanism of the retinoids-mediated growth inhibition.

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고온성 알콜발효 효모의 Alcohol Dehydrogenase의 특성

  • 예상수;임시규;손호용;진익렬;이인구;김영호;서정훈;박완
    • 한국미생물·생명공학회지
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    • 제25권4호
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    • pp.386-390
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    • 1997
  • The characteristics of alcohol dehydrogenase (ADH, EC 1.1.1.1, alcohol:NAD oxidoreductase) of thermotolerant alcohol-producing yeasts, Saccharomyces cerevisiae RA-74-2 and Kluyveromyces marxianus RA-912, were compared with that of mesophilic S. cerevisiae D, an industrial strain. Under anaerobic culture condition, both S. cerevisiae RA-74-2 and D had similar level of ADH activity at 30$\circ$C, and the activity of S. cerevisiae RA-74-2 at 37$\circ$C was the same level at 30$\circ$C. However, the level of ADH activity of S. cerevisiae D at 37$\circ$C decreased about 70% of that at 30$\circ$C. The level of enzyme activity of K. marxianus RA-912, which showed lower alcohol productivity than S. cerevisiae RA-74-2 and D, was about 43% of those strains at 30$\circ$C, and decreased somewhat at 37$\circ$C. The results showed a good correlation between the alcohol productivities and the level of ADH activities of these strains grown at 30$\circ$C and 37$\circ$C. And the higher heat stability of ADH of S. cerevisiae RA-74-2 than that of S. cerevisiae D seemed to reflect the ability of high temperature fermentation. Despite of its fermentation ability even at 45$\circ$C, however, the ADH of K. marxianus RA-912 showed lower heat stability than that of S. cerevisiae D. Both S. cerevisiae RA-74-2 and D showed similar patterns of two bands of ADH isozyme, and the low band of S. cerevisiae RA-74-2 moved slightly faster than that of S. cerevisiae D. The staining intensity of the bands of S. cerevisiae D at 37$\circ$C was weaker than those at 30$\circ$C. However, S. cerevisiae RA-74-2 showed no differences in total intensity of the bands of 30$\circ$C and 37$\circ$C. As the patterns of cellular proteins and ADH isozyme of K. marxianus RA-912 were different from S. cerevisiae RA-74-2 and D, K. marxianus might have its own characteristic ADH system.

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F9 EC 세포에서 레티노산에 의해 유도되는 Hoxc 유전자의 발현에 히스톤 메틸화가 미치는 영향 (Histone Methylation Regulates Retinoic Acid-induced Hoxc Gene Expression in F9 EC Cells)

  • 민혜현;김명희
    • 생명과학회지
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    • 제25권6호
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    • pp.703-708
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    • 2015
  • Hox 유전자는 호메오도메인을 포함한 전사인자로써, 발생 과정 중 전후축을 따라 몸의 형태 형성을 조절하는 역할을 한다. 레티노산(RA)은 발생 과정에서 필수적인 형태형성인자이며 세포의 특성을 결정하는데 중요한 조절자이다. 특히, RA는 생쥐나 인간으로부터 만들어진 배아암종(EC)세포에서 Hox 유전자의 발현을 조절한다고 밝혀져 있다. 또한 RA에 의한 세포 분화와 유전자 조절 과정에 히스톤 변이가 중요한 역할을 하는 것으로 보고되어 있다. 히스톤 변이가 RA에 의해 유도되는 Hox 유전자의 발현에 특이적인 역할을 할 것으로 유추되기 때문에, 이 연구의 목적은 F9 생쥐배아 기형암종세포에서 RA에 의해 유도되는 Hoxc 유전자의 순차적인 발현이 히스톤 변이에 의해 일어나는 것인지를 조사하는 것이다. Hox 유전자의 발현 양상과 히스톤 변이는 semi-quantitative RT-PCR, RNA-sequencing과 chromatin immuno-precipitation (ChIP)-PCR 기법을 이용하여 관찰하였다. RA 처리 후(0일(D0), 1일(D1), 3일(D3)), Hoxc4 유전자의 발현(D1)은 Hoxc5부터 –c10 유전자(D3)보다 먼저 시작되었다. Hox가 발현하지 않는 D0 샘플은 전사 억제 마커인 H3K27me3이 모든 Hoxc 좌위에 강하게 표지 되어 있었으나 D1과 D3 샘플에서는 모든 좌위의 H3K27me3 표지가 확연히 줄어들어 있었다. 전사 발현 마커인 H3K4me3가 Hoxc 유전자의 순차적인 발현과 더 연관성이 있는 것으로 보이는데 D1에서 Hoxc4 발현과 함께 H3K4me3이 표지 되어 있었고, D3에서는 Hoxc 유전자 발현과 함께 모든 좌위에서 H3K4me3 마커가 존재했기 때문이다. 모든 결과를 종합해 보았을 때 F9 세포에서 RA에 의해 유도된 Hoxc 유전자의 순차적인 발현은 Hoxc 좌위에서 H3K27me3가 사라지고, H3K4me3가 표지 되는 히스톤 메틸화의 변이에 의해 결정되는 것으로 사료된다.

Bacillus Subtilis KJ-3를 이용한 생물전환물 및 그 혼합물의 생리활성 (Physiological Activities of Bioconversion Products Using Bacillus Subtillis KJ-3 and Their Mixtures)

  • 이진영;동재경;정유성;김미령;강재선
    • 생명과학회지
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    • 제29권10호
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    • pp.1086-1095
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    • 2019
  • 이 연구는 Red allium cepa (RA), Cucurbita moschata Duch (CM), and Angelica gigas Nakai (AG)의 혼합물을 이용한 새로운 기능성 물질을 개발하기 위해 수행되었다. RA와 CM은 수분 함량이 높아 저장 안정성이 매우 낮다. 따라서 각 재료들의 주요성분들을 추출하여 함량 분석 후 연구에 사용하였다. RA는 에탄올 추출 후 Bacilus subillis KJ-3 (BS3)에 의해 생물전환하였다. 생물전환 후, RA의 Quercetin 함량은 128.9% 증가되었음을 확인하였다. CM의 에탄올 추출물에서는 ${\beta}$-카로틴을 검출하였고 함량은 0.2 mg/g으로 낮았다. AG 에탄올 추출물(1 mg)의 데커신 및 데커시놀 엔젤레이트(D/DA)는 각각 0.4146 mg과 0.3659 mg을 함유했다. D/DA의 순도는 약 78%로 나타났다. 이들 각각의 물질 및 혼합물(혼합물 1 (RA:CM:AG = 5:2:3), 혼합물 2 (RA:CM: AG = 3:5:2), 혼합물 3 (RA:CM:AG = 3:2:5)의 총 플라보노이드 함량과 폴리페놀 함량을 측정하였다. 세포 생존율, 항염증 활성, 항산화 능력 또한 평가하였다. 모든 결과를 종합하여 볼 때, 혼합물 3 (RA:CM:AG=3:2:5)의 항산화 작용이 가장 효과적이었다. 따라서 이러한 연구 결과는 향후 RA, CM, AG의 3:2:5 혼합물을 이용한 식품개발을 위한 기초자료로 활용하고자 한다.

Electrochemical Ionic Mass Transfer Correlation in Fluid-Saturated Porous Layer

  • Cho, Eun Su
    • Korean Chemical Engineering Research
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    • 제53권6호
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    • pp.814-817
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    • 2015
  • A new ionic mass transfer correlation is derived for the fluid-saturated, horizontal porous layer. Darcy-Forchheimer model is used to explain characteristics of fluid motion. Based on the microscales of turbulence a backbone mass transfer relation is derived as a function of the Darcy-Rayleigh number, $Ra_D$ and the porous medium Schmidt number, $Sc_p$. For the Darcy's limit of $Sc_p{\gg}Ra_D$, the Sherwood number, Sh is a function of $Ra_D$ only. However, for the region of high $Ra_D$, Sh can be related with $Ra_DSc_p$. Based on the present backbone equation and the electrochemical mass transfer experiments which are electro plating or electroless plating, the new ionic mass transfer correlation is suggested in the porous media.

A study on F8L10D-N LoRa RF Module for Drone Based live Broadcasting system

  • Mfitumukiza, Joseph;Mariappan, Vinayagam;Lee, Minwoo;Cho, Juphil;Cha, Jaesang
    • International Journal of Advanced Culture Technology
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    • 제4권4호
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    • pp.1-5
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    • 2016
  • In this paper, we present the study on the proposed design of a real-time transmission of a video from the drone to broadcasting station (OBVan) by using F8L10D-N LoRa Module. Nowadays, LoRa technology is proved to be the mass of low cost, long range machine-to-machine connectivity. Particularly in the field of broadcasting and communication system, F8L10D-N LoRa RF Module spread spectrum technology with long transmission distance and strong penetrative ability that is double stronger than traditional FSK as well as PSK modulation scheme.

레틴알 안정화를 위한 사이클로덱스트린-리포좀에 관한 연구 (Study on Stabilization of Retinaldehyde using Drug-in-Cyclodextrinin-Liposome (DCL) for Skin Wrinkle Improvement)

  • 하지훈;최형;홍인기;한상근;빈범호
    • 대한화장품학회지
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    • 제48권1호
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    • pp.77-85
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    • 2022
  • 레틴알(RA)은 레티놀과 레티노익애씨드의 중간체로 비타민A 유도체이며 주름개선 효과가 우수하다. 본 연구에서는 drug-in-cyclodextrin-in-liposome (DCL)을 이용하여 레틴알의 안정성을 높였다. 레틴알과 hydroxypropyl-β-cyclodextrin (HP-β-CD) 복합체를 동결건조 방식으로 제조하였고, UV-Vis 분광법, FT-IR 및 SEM 이미지로 레틴알의 포접 여부를 확인하였다. 레틴알과 HP-β-CD의 비율이 1 : 15 (w/w)일 때 약 95.6% 포집되었다. 레틴알-HP-β-CD 복합체는 호모믹서 및 마이크로플루다이저로 리포좀에 담지시켰으며, 평균 입자 크기는 215.3 ± 4.2 nm, 제타포텐셜 -33.2 ± 1.5 mv로 나타났다. 레틴알의 분해 안정도 평가에서, 물에서 레틴알-HP-β-CD-리포좀의 레틴알 감소율은 1.8%로 레틴알-리포좀(5.8%), 레틴알-HP-β-CD복합체(9.7%), 레틴알 단독(37.6%)보다 높게 나타났다. 레틴알-HP-β-CD-리포좀이 함유된 크림(0.05% RA 함유)을 제조하여, 미간, 이마, 목, 눈가, 입가, 팔자 주름개선 효능 및 피부 치밀도를 2 ~ 4 주간 평가하였다. 그 결과 레틴알크림은 피부 자극 없이 유의한 주름 개선 효과를 보였다. 결론적으로, DCL시스템을 이용한 이중 안정화 기술은 레틴알의 안정화를 높여 피부 주름 개선 효과에 기여함을 확인하였다.

Cytochalasin D Regulates Retinoic Acid Induced COX-2 Expression but not Dedifferentiation via p38kinase Pathway in Rabbit Articular Chondrocytes

  • ;김송자
    • 대한의생명과학회지
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    • 제15권4호
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    • pp.343-347
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    • 2009
  • Cytochalasin D (CD) is known as a disruptor of actin cytoskeleton architecture in chondrocytes. We have studied the role of CD in retinoic acid (RA) caused dedifferentiation and inflammation responses in rabbit articular chondrocytes. We have examined the effect of CD on RA induced dedifferentiation of chondrocytes. CD inhibited RA induced dedifferentiation determined by Western blot analysis and Alcian blue staining in rabbit articular chondrocytes. Also, CD additionally reduced inflammation response molecules such as cyclooxygenase-2 (COX-2) and prostaglandin $E_2$ ($PGE_2$) in RA treated cells. Treatment of CD reduced phosphorylation of p38 by treatment of RA. Inhibiton of p38kinase with SB203580 reduced expression of COX-2 and production of $PGE_2$ by treatment of CD in RA treated cells. But, Inhibiton of p38kinase with SB203580 did not any relationship with effect of CD on RA caused dedifferentiation. In summary, our results indicate that CD regulates RA reduced expression of COX-2 and production of PGE2 via p38kinase pathway.

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All-trans Retinoic Acid-Associated Low Molecular Weight Water-Soluble Chitosan N anoparticles Based on Ion Complex

  • Kim Dong-Gon;Choi Changyong;Jeong Young-Il;Jang Mi-Kyeong;Nah Jae-Woon;Kang Seong-Koo;Bang Moon-Soo
    • Macromolecular Research
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    • 제14권1호
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    • pp.66-72
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    • 2006
  • The purpose of this study is to develop novel nanoparticles based on polyion complex formation between low molecular weight water-soluble chitosan (LMWSC) and all-trans retinoic acid (atRA). LMWSC nanoparticles encapsulating atRA based on polyion complex were prepared by mixing of atRA into LMWSC aqueous solution using ultrasonication. In FTIR spectra, the carbonyl group of atRA at 1690 $cm^{-1}$ disappeared or decreased when ion complexes were formed between LMWSC and atRA. In ${1}^H$ NMR spectra, specific peaks of atRA disappeared when atRA-encapsulated LMWSC (RAC) nanoparticles were reconstituted into $D_{2}O$ while specific peaks both of atRA and LMWSC appeared in $D_{2}O$/DMSO (1/3, v/v) mixture. XRD patterns also showed that the crystal peaks of atRA were disappeared by encapsulation into LMWSC nanoparticles. LMWSC nanoparticles encapsulating atRA have spherical shapes with particle size below 200 nm. The mechanism of encapsulation of atRA into LMWSC nanoparticles was thought to be an ion complex formation between LMWSC and atRA. LMWSC nanoparticles showed high atRA loading efficiency over 90$\%$ (w/w). AtRA was continuously released from nanoparticles over 10 days. In in vitro cell cytotoxicity test, free atRA showed higher cytotoxic effect against CT 26 colon carcinoma cell line on 1 day. However, RAC nanoparticles showed similar cytotoxicity against CT 26 cells on 2 day. These results suggest the potential for the introduction of LMWSC nanoparticles into various biomedical fields such as drug delivery.